Comammox Nitrospira among dominant ammonia oxidizers within aquarium biofilter microbial communities.

Nitrification by aquarium biofilters transforms ammonia waste (NH3/NH4+) to less toxic nitrate (NO3-) via nitrite (NO2-). Prior to the discovery of complete ammonia-oxidizing ("comammox" or CMX) Nitrospira, previous research revealed that ammonia-oxidizing archaea (AOA) dominated over ammonia-oxidizing bacteria (AOB) in freshwater aquarium biofilters. Here, we profiled aquarium biofilter microbial communities and quantified the abundance of all three known ammonia oxidizers using 16S rRNA gene sequencing and quantitative PCR (qPCR), respectively. Biofilter and water samples were each collected from representative residential and commercial freshwater and saltwater aquaria. Distinct biofilter microbial communities were associated with freshwater and saltwater biofilters. Comammox Nitrospira amoA genes were detected in all 38 freshwater biofilter samples (average CMX amoA genes: 2.2 × 103 ± 1.5 × 103 copies/ng) and dominant in 30, whereas AOA were present in 35 freshwater biofilter samples (average AOA amoA genes: 1.1 × 103 ± 2.7 × 103 copies/ng) and only dominant in 7 of them. The AOB were at relatively low abundance within biofilters (average of 3.2 × 101 ± 1.1 × 102 copies of AOB amoA genes/ng of DNA), except for the aquarium with the highest ammonia concentration. For saltwater biofilters, AOA or AOB were differentially abundant, with no comammox Nitrospira detected. Additional sequencing of Nitrospira amoA genes revealed differential distributions, suggesting niche adaptation based on water chemistry (e.g., ammonia, carbonate hardness, and alkalinity). Network analysis of freshwater microbial communities demonstrated positive correlations between nitrifiers and heterotrophs, suggesting metabolic and ecological interactions within biofilters. These results demonstrate that comammox Nitrospira plays a previously overlooked, but important role in home aquarium biofilter nitrification. IMPORTANCE: Nitrification is a crucial process that converts toxic ammonia waste into less harmful nitrate that occurs in aquarium biofilters. Prior research found that ammonia-oxidizing archaea (AOA) were dominant over ammonia-oxidizing bacteria (AOB) in freshwater aquarium biofilters. Our study profiled microbial communities of aquarium biofilters and quantified the abundance of all currently known groups of aerobic ammonia oxidizers. The findings reveal that complete ammonia-oxidizing (comammox) Nitrospira were present in all freshwater aquarium biofilter samples in high abundance, challenging our previous understanding of aquarium nitrification. We also highlight niche adaptation of ammonia oxidizers based on salinity. The network analysis of freshwater biofilter microbial communities revealed significant positive correlations among nitrifiers and other community members, suggesting intricate interactions within biofilter communities. Overall, this study expands our understanding of nitrification in aquarium biofilters, emphasizes the role of comammox Nitrospira, and highlights the value of aquaria as microcosms for studying nitrifier ecology.

Cutaneous infection due to Mycobacterium marinum: A series of four cases from Kerala, India.

Mycobacterium marinum is a non-tuberculous mycobacterium which can be found in naturally occurring, non-chlorinated water sources and is a known pathogen that affects fish. In humans, M. marinum typically results in cutaneous lesions, it can occasionally lead to more invasive disorders. We discuss four cases of M. marinum-related cutaneous infections examined in a tertiary care facility. We want to draw attention to the challenges of accurately diagnosing this infection, stress the significance of having a high level of clinical suspicion in order to identify it, and discuss the available treatment choices.

Ocular injury from saltwater coral palytoxin.

We describe a case of a young male who presented to the emergency department with unilateral eye pain, blurred vision, conjunctival injection, and ocular pH of 9, one day after direct ocular exposure to palytoxin (PTX) from coral in a home saltwater fish tank. Although uncommon, ocular PTX toxicity is a potentially vision-threatening condition that requires prompt recognition. This case report documents the successful management of presumed ocular PTX exposure and suggests additional workup and treatment considerations for future patients.

Diversity and distribution of Aphanomyces astaci in a European hotspot of ornamental crayfish introductions.

Ornamental trade has become an important introduction pathway of non-native aquatic species worldwide. Correspondingly, there has been an alarming increase in the number of established crayfish of aquarium origin in Europe over the previous decade. The oomycete Aphanomyces astaci, the pathogen causing crayfish plague responsible for serious declines of European crayfish populations, is dispersed with introduced North American crayfish. The role of ornamental taxa in introducing and spreading different genotypes of this pathogen in open waters remains unclear. We investigated the distribution, prevalence, and diversity of A. astaci in Budapest, Hungary, which became a hotspot of aquarium crayfish introductions. Their establishment in this area was facilitated by locally abundant thermal waters. We screened for A. astaci in six host taxa from 18 sites sampled between 2018 and 2021: five cambarids (Cambarellus patzcuarensis, Faxonius limosus, Procambarus alleni, P. clarkii, P. virginalis) and one native astacid (Pontastacus leptodactylus). The pathogen was confirmed at five sampled sites in four host taxa: P. virginalis, P. clarkii, F. limosus, and for the first time in European open waters also in P. alleni. Genotyping was successful only in individuals from two different brooks where multiple host species coexisted but revealed unexpected patterns. Mitochondrial B-haplogroup of A. astaci, previously usually reported from Pacifastacus leniusculus or infected European species, was detected in P. virginalis at both sites, and in both F. limosus and P. virginalis sampled from a thermally stable tributary of Barát brook in 2018. In contrast, A-haplogroup of A. astaci was detected in coexisting F. limosus, P. virginalis and P. clarkii sampled in the same watercourse just a few hundred meters downstream in 2020. Additional genotyping methods indicated that a previously unknown A. astaci strain was associated with the latter haplogroup. One P. virginalis individual from 2020 was apparently co-infected by strains representing both mitochondrial haplogroups. The results indicated multiple sources of A. astaci in Budapest, likely directly associated with the introduction of ornamental species, interspecific transmission of this pathogen among ornamental hosts, and potential for a quick spatial or temporal turnover of dominant A. astaci strains at a certain locality. This highlights that in regions with high richness of potential A. astaci hosts, host taxon/pathogen genotype combinations become unpredictable, which might prevent reliable genotyping of pathogen sources in local crayfish mass mortalities.

Life in a fishbowl: Space and environmental enrichment affect behaviour of Betta splendens.

The public has expressed growing concern for the well-being of fishes, including popular pet species such as the Siamese fighting fish (Betta splendens). In captivity, male Bettas behave aggressively, often causing injuries and death if housed together. As a result, they are typically isolated in small fishbowls, which has been widely criticised as cruel. To investigate the impact of keeping Bettas in these conditions, we recorded the behaviour of individual males in containers of different sizes that were either bare or enriched with gravel, large rocks, and live plants. When male Bettas were housed individually in small bowls (0.5 L) they spent less time swimming than they did when they were kept in larger aquaria (10, 38, and 208 L). Fish that were kept in enriched containers exhibited more instances of swimming. To determine if two male Bettas housed together might coexist peacefully if given enough space and cover from plants and large rocks, we quantified the behaviour of pairs of male Bettas in bare or enriched aquaria of different sizes (10, 38, 208, 378 L). Fish performed fewer approaches and aggressive displays, but not attacks, and more bouts of foraging, when in larger aquaria. This study shows that the small fishbowls typically used in pet stores suppress swimming behaviour in male Bettas and at least a 10-L aquarium is required to ensure full expression of swimming behaviour. Furthermore, even the use of very large aquaria cannot guarantee peaceful cohabitation between two males.

Husbandry conditions of spotted ratfish (Hydrolagus colliei, Chimaeriformes) in aquaria for successful embryonic development and long-term survival of juveniles.

The spotted ratfish Hydrolagus colliei is the most common holocephalan species exhibited in aquaria worldwide for introducing deep-sea environments and raising awareness of their conservation. However, little is known about the biology of H. colliei. Current practices in aquaria allow long-term survival of sexually mature H. colliei specimens; however, this species struggles to complete a reproductive cycle in captivity mostly because embryos do not reach the hatchling stage. The aquarists of Planet Ocean Montpellier (POM, France) have bred H. colliei for 15 years and recorded parameters suitable for this species' successful embryonic and post-embryonic development. POM aquarists now regularly record egg-laying events of H. colliei and use four tanks to incubate eggs and raise neonates, late hatchlings, early and intermediate juveniles, subadults, and sexually mature specimens. In this work we provide the first long-term biometric data on H. colliei from the hatchling to the subadult stage. We also report the biotic and abiotic parameters sufficient to breed H. colliei in aquaria. We finally describe the methods used to facilitate individual monitoring of specimens along the ontogeny and several pathologies identified in this species, their putative causes, and the corresponding treatments. This work highlights the importance of ex situ research and points to the valuable outcomes of collaborative efforts between aquaria and academia in deciphering the biology of species whose study in the wild remains challenging.

Traumatic telson avulsion and attempted external stabilization in Atlantic horseshoe crabs (Limulus polyphemus).

Atlantic horseshoe crabs (Limulus polyphemus) are prevalent in public aquarium touch pools. Despite their popularity, the literature concerning medical management under managed care is sparse. Noninfectious conditions include trauma to the exoskeleton and compound eyes; however, injury to the soft tissue connecting the telson and opisthosoma has not been previously reported. This report describes telson avulsion in three Atlantic horseshoe crabs at a public aquarium and attempted external stabilization with a vascular silicone tie in two of three affected animals. The horseshoe crab that received no veterinary intervention suffered a complete telson amputation 1 month after the initial injury. Although the two other horseshoe crabs did well postoperatively, and external stabilization prevented further avulsion or amputation, they still could not right themselves if flipped into dorsal recumbency. All three horseshoe crabs were ultimately euthanized due to animal welfare concerns. To avoid potentially serious telson ligament injury, horseshoe crabs should never be picked up by the telson. The authors recommend that telson ligament injuries be addressed promptly, any inciting cause be mitigated, and external stabilization be considered earlier.

Nephrolithiasis in a giant Pacific octopus Enteroctopus dofleini.

Managed care of cephalopods can be complicated by numerous factors including infectious and non-infectious disease, environmental stressors, and anatomic and physiological changes associated with senescence. The current report describes a unique case of nephrolithiasis in a senescent, >2 yr old female Pacific octopus Enteroctopus dofleini housed in a public aquarium. Clinical signs included generalized external pallor, inappetence progressing to anorexia, lethargy, and a slow healing mantle abrasion over a period of a year. Due to the animal's declining condition, humane euthanasia was elected. At necropsy, multiple, small, approximately 1-5 mm diameter crystalline deposits were reported throughout all sections of the renal appendages. Histopathology identified a large crystal expanding and rupturing a focal tubule, causing necrosis, ulceration, and hemocytic infiltration. Crystalline stone analysis revealed that the nephrolith was composed of 100% ammonium acid urate. Additional changes to the digestive gland included marked atrophy with fibrosis correlated to the animal's history of hyporexia/anorexia secondary to senescence. To our knowledge, this is the first report of nephrolithiasis in E. dofleini.

Core single nucleotide polymorphism analysis reveals transmission of Mycobacterium marinum between animal and environmental sources in two aquaria.

Mycobacterium marinum is a slow-growing, photochromogenic nontuberculous mycobacterium, which can cause mycobacteriosis in various animals, including humans. Several cases of fish mycobacteriosis have been reported to date. Mycobacterium marinum has also been isolated from aquatic environmental sources such as water, sand, biofilms, and plants in the natural environments. Hence, we hypothesized that a wide variety of sources could be involved in the transmission of M. marinum. In this study, we tested this hypothesis by isolating M. marinum from various sources such as fish, invertebrates, seagrass, periphytons, biofilms, sand, and/or water in two aquaria in Japan and conducting a phylogenetic analysis based on single-nucleotide polymorphisms (SNPs) using whole-genome sequences of the isolated strains. The analysis revealed that the strains from animal and environmental sources belonged to the same clusters. This molecular-based study epidemiologically confirmed that various sources, including fish, invertebrates, and environmental sources, could be involved in transmission of M. marinum in a closed-rearing environment. This is the first report where M. marinum was isolated from different sources, and various transmission routes were confirmed in actual cases, which provided essential information to improve the epidemiology of M. marinum.

The environmental impact of keeping a tropical aquarium in Northern Europe.

Tropical fishkeeping is a popular practice in societies across the globe and involves recreating and sustaining an entire ecosystem in an aquarium within a domestic setting. The process invariably has an environmental impact, yet an assessment of this impact has previously been limited to the ecological consequences of harvesting fish from the wild or the release of non-native fish species. Provided here are the first estimates of carbon dioxide equivalent (CO2 eq) emissions produced from running a tropical aquarium across multiple countries in Northern Europe (France, Poland and the UK), along with water consumption. Estimates were produced in silico and are discussed in the context of freshwater and marine aquariums, calculated using example aquarium sizes of 50, 200 and 400 l. Using estimates from the UK, depending on size and running conditions, a tropical aquarium produces an estimated 85.3-635.2 kg of CO2 eq per year, equating to 1.6%-12.4% of the UK annual average household CO2 emissions, and uses 156-31,200 l of water per year, equating to 0.2%-30.1% of the UK annual average household water usage. Despite this, comparison with the CO2 eq of an average-size dog (127-1592 kg of CO2 eq per year) or cat (121-251 kg of CO2 eq per year), estimated from meat consumption alone, demonstrates that ornamental fishkeeping can be a more environmentally conscious pet choice. In addition, the majority of CO2 eq produced from tropical fishkeeping is generated from the energy consumption of aquarium equipment and as more national electricity grids begin to decarbonize, this estimate should decrease.

Characteristics and Risk Assessment of Microplastics in Water and Mussels Sampled from Cape Town Harbour and Two Oceans Aquarium, South Africa.

The aim of this study was to measure the characteristics and risk assessment of microplastics (MPs) in Cape Town Harbour (CTH) and the Two Oceans Aquarium (TOA) in Cape Town, South Africa from 2018 to 2020. Water and mussel MP samples were analyzed at 3 sites in CTH and TOA, respectively. Microplastics were mainly filamentous, black/grey and 1000-2000 µm in size. A total of 1778 MPs, averaging 7.50 (± 0.6 standard error of the mean, SEM) MPs/unit were recorded. Average MP concentrations were 10.3 ± 1.1 MPs/L in water and 6.27 ± 0.59 MPs/individual or, based on weight, 3.05 ± 1.09 MPs/g soft tissue wet weight in mussels. Average MPs in seawater in CTH (12.08 ± 1.3 SEM MPs/L) was significantly higher (4.61 ± 1.1 MPs/L) than inside the TOA (U = 536, p = 0.04). Various risk assessment calculations indicate that MPs in seawater poses a greater ecological risk than MPs in mussels at the sites sampled.

Ultraviolet light alters experimental aquarium water microbial communities.

The effect of ultraviolet (UV) light exposure, alone and in combination with CO2 exposure, on the water microbial community composition was tested in replicate experimental aquaria using source water from an established Amazon-themed exhibit housing mixed species of fishes. Total bacterial abundance, α-diversity metrics, and ß-diversity metrics were determined 3 weeks and 1 week before, and weekly during 8 weeks of continuous treatment. The UV treatment significantly lowered the overall bacterial abundance while CO2 treatment had no effect. However, the UV exposure effect was variable across phyla. Some phyla were decreased while others were increased, including some of potential clinical significance. At the genus level, there were no significant differences in the relative abundance of Mycobacteria between treatments and an increase in the relative abundance of Aeromonas spp. with UV light treatment. Further work is needed to determine if the observed effects are dose-dependent or if different exposure doses produce different results.

Tandem spawning and development in the African freshwater pufferfish Tetraodon schoutedeni.

To advance breeding techniques for the African freshwater pufferfish Tetraodon schoutedeni and observe tandem spawning closely, we monitored the reproduction of captive individuals. Eight spawning sessions (stable water temperature 24-25°C; daily light period 07:00-19:00) occurred between May 2016 and November 2017. After 65-150 min of tandem swimming (the male biting and clinging to the female's abdomen), 3-50 spherical, weakly adhesive eggs were spawned, being scattered onto the sandy substrate or water plants. The removal of cohabitants (potentially eating spawned eggs) and provision of small initial food items, such as small-type Brachionus spp., for larval fish were essential for successful breeding.

Food-related substrate preference in juveniles seastar Echinaster (Othilia) brasiliensis (Müller & Troschel,1842) in captivity.

There are only a few studies that describe the larval development of Echinaster or aspects on culture systems for the genus. For starfishes, the choice of suitable substrates has received special attention since it could influence the acid-base balance of the water, movement capacity and predation rate. The objective of this study was to evaluate the ideal food-related substrate for the rearing of juvenile Echinaster brasiliensis. A batch of fertilized eggs released in spontaneous spawning was collected and kept in a plankton-kreisel until metamorphosis. Data on preference of food-related substrate was recorded for 10 weeks from day 58 post-release. From release to 132 days old, arm length increased from 0.81 mm to 1.31 ± 0.03 mm. Considering the sudden increase in arm length (AL), it was estimated that feeding started around 40 days of age. Regarding food-related substrate preferences, biofilm grown on "rocks" showed a significant difference among other treatments, adding up to 50% of preference (p < .05). For sponge and biofilm from bio media, there was no statistical difference for the whole period. In this study, sponges showed to be the least preferred food-related substrate for post-settlement juveniles. Considering that Echinaster and other starfish are commonly maintained on a diet of collected or cultured sponges, difficulties in sourcing a ready supply throughout the year represent limitations to their sole use within commercial or laboratory-scale production. In this sense, the use of biofilm from biological media for the feeding of juvenile starfish is not yet reported in the literature and showed to be an easy and promising option.

Increasing the data on elasmobranch plasma protein electrophoresis: electrophoretogram reference values determination in the undulate skate (Raja Undulata) and the nursehound shark (Scyliorhinus stellaris) maintained under human care.

BACKGROUND: This study determined plasma protein electrophoresis (PPE) reference intervals in two elasmobranch species: the undulate skate (Raja undulata) and the nursehound shark (Scyliorhinus stellaris), using a reference population of 48 undulate skates (27 males, 21 females) and 62 nursehounds (32 males, 30 females), considered to be clinically healthy. Plasma samples were analyzed using capillary zone electrophoresis (CZE). RESULTS: The undulate skate electrophoretogram resembled those previously reported in other batoids and could be divided into seven consistent fractions. No statistically significant differences were detected between sexes and developmental stages. The nursehound electrophoretogram was similar to that previously described in other shark species and could be divided into eight consistent fractions. Fraction 5% was significantly higher in juvenile nursehounds when compared to adults, while fraction 6 concentration and percentage were significantly higher in adults. Fraction 4% was higher in males than in females. Albumin band was not detected, and pre-albumin was negligible in both studied species. Alpha-globulins were predominant in the undulate skate, while beta-globulins were predominant in nursehounds. Statistically significant differences were found in all electrophoretogram fraction percentages and concentrations between the two species. CONCLUSION: To the authors knowledge, this is the first study reporting PPE values in undulate skates and nursehounds, and the first study using CZE in elasmobranch plasma. These findings can serve as a primary reference for health monitoring in both species and will add to the limited data available on PPE in elasmobranchs.

Autochthonous transmission of the Indomalayan parasite, Transversotrema patialense, in the Caribbean: Molecular, morphological, and experimental evidence.

The Asian freshwater snail Melanoides tuberculata has been established since the 1960s in the Americas, where it transmits cercariae of a small number of digenetic trematode species from its native range. In 2021-2022, 24 M. tuberculata were discovered shedding transversotrematid cercariae in Puerto Rico, where parasites of this snail have not been previously studied. Adult transversotrematids (in some cases, gravid) were found on field-caught fish and on fish exposed to shedding snails, including on fish species native to Puerto Rico. Adults and cercariae were identified as Transversotrema patialense (Soparkar, 1924), a species native to the Indomalayan region. Morphological identification was supported with 28S rDNA sequences closely matching that from unidentified transversotrematid cercariae in Thailand. The absence of T. patialense in snails collected prior to 2021, increasing prevalence of infection in snails collected thereafter, and lack of variation in parasite DNA sequences (28S, internal transcribed spacer 2, cytochrome c oxidase I) from three isolates are consistent with a recently introduced and possibly expanding parasite population. Transversotrema patialense has been recorded outside its native range before, but most studies (including a prior record in the Americas) reported the parasite from captive hosts from commercial sources such as pet shops. The present results thus provide the first demonstration of natural transmission of T. patialense in the Americas. Phylogenetic analysis of 28S but not of ITS2 show the transversotrematid genus Transversotrema Witenberg, 1944 is paraphyletic, with Crusziella Cribb, Bray and Barker 1992 nested within it.

Noise-Robust Multimodal Audio-Visual Speech Recognition System for Speech-Based Interaction Applications.

Speech is a commonly used interaction-recognition technique in edutainment-based systems and is a key technology for smooth educational learning and user-system interaction. However, its application to real environments is limited owing to the various noise disruptions in real environments. In this study, an audio and visual information-based multimode interaction system is proposed that enables virtual aquarium systems that use speech to interact to be robust to ambient noise. For audio-based speech recognition, a list of words recognized by a speech API is expressed as word vectors using a pretrained model. Meanwhile, vision-based speech recognition uses a composite end-to-end deep neural network. Subsequently, the vectors derived from the API and vision are classified after concatenation. The signal-to-noise ratio of the proposed system was determined based on data from four types of noise environments. Furthermore, it was tested for accuracy and efficiency against existing single-mode strategies for extracting visual features and audio speech recognition. Its average recognition rate was 91.42% when only speech was used, and improved by 6.7% to 98.12% when audio and visual information were combined. This method can be helpful in various real-world settings where speech recognition is regularly utilized, such as cafés, museums, music halls, and kiosks.

Unique bacterial communities associated with components of an artificial aquarium ecosystem and their possible contributions to nutrient cycling in this microecosystem.

In order to better understand the bacterial distribution characteristics in a whole microecosystem, the bacterial communities in different components of an artificial aquarium (i.e., plants, fishes, sand and water) were characterized using high throughput sequencing of bacterial 16S rRNA genes. Across all samples, 2873 operational taxonomic units were identified and assigned to 771 genera in 36 phyla. In a principle coordinate analysis, samples clustered according to their origin, indicating that bacterial communities from the same component were most similar. Further taxonomic analysis revealed that most dominant genera, even those with the similar functions, were biased to one component: Nitrospira and Rhodobacter were mainly abundant in plant samples; Rhodococcus, Serratia, Ralstonia, Sphingobacterium and Pseudomonas were most common in sand samples; Cetobacterium and Aeromonas dominated fish samples; and Flavobacterium, Alpinimonas and Limnobacter were especially common in water samples. Functional predictions performed by PICRUSt and the dominant genera exhibited that bacteria detected in each component could participate in all nutrient cycles in the aquarium. However, those involved in carbon and nitrogen cycling were most common in plant and fish samples, while phosphate metabolism-related pathways were more abundant in sand and water samples. Moreover, the aquarium plants, in association with their bacterial communities might be the most important component in the aquarium, as indicated by their highest bacterial richness and diversity. This study adds to our understanding on the differences in the microbiome of different components and their possible contributions to nutrient cycling in a self-sustaining aquarium.

Estimating the total length of Mekong giant catfish, Pangasianodon gigas, in an aquarium via stereo-video shooting and direct linear transformation.

The Mekong giant catfish (MGC), Pangasianodon gigas, is one of the world's largest catfish species. Endemic to the Mekong River system, the MGC is critically endangered. Six MGC were transported from Thailand to the Gifu World Freshwater Aquarium (GWFA) in Japan in May 2004. The MGC have not been measured since they were introduced to the GWFA due to the challenges associated with their large size. Traditional methods, such as anesthesia and net-capture, could harm the fish, potentially reducing the population further. However, understanding the ecology of the species is essential to conservation efforts. Seasonal feeding rhythms and long-term fasting were previously observed in captive MGCs. To investigate the effect of long-term fasting on MGC growth, total length measurements are necessary. In this study, we applied a noncontact method to estimate the total length via image analysis. We shot a stereo-video of free-swimming MGC in the aquarium tank using two digital video cameras and analyzed the 3D images using the direct linear transformation method. We successfully estimated the total length of each MGC individual without contacting the fish. This accurate estimation method is versatile, simple, and useful in aquarium breeding and is also recommended from the viewpoint of animal welfare because it is a noninvasive method of measurement.

Conservation psychology and the legacy of Carol Saunders for zoo and aquarium programs, networks, practices, and exhibits.

Conservation Psychology created a dialog between environmental conservation and behavioral sciences. With an outsized influence by Dr. Carol Saunders, it started at Brookfield Zoo in Chicago exploring questions about the impacts of a zoo visit, and particularly how human behavior influences environmental outcomes for our planet. Here we explore how Conservation Psychology influenced the development of programs, exhibits and communities of practice at Brookfield Zoo and elsewhere in the zoo and aquarium world, and how eventually these applications changed the way modern zoos and aquariums operate. We present testimonials and review a handful of examples in which Conservation Psychology led to tangible programs, practices and wide professional networks at zoos and aquariums. These include an exploration of the future of zoos with George Rabb, followed by the legacy of nature play and the groundbreaking Hamill Family Play Zoo. Furthermore, we discuss how visitor studies at zoos and aquariums were influenced by Conservation Psychology, including the development of the Association of Zoos and Aquariums Social Science Research and Evaluation Scientific Advisory Group and two climate change education networks. We end with the development of tools, practices, and professional networks to explore empathy for animals. Most of these programs were envisioned or facilitated by Dr. Saunders, who was always a role model with an impact and a legacy that lives on.