Molecular detection of Sarcocystis sp. in a kept under human care black capuchin monkey (Sapajus nigritus., Goldfuss 1809) with necrotizing encephalitis.

A senile male black capuchin monkey (Sapajus nigritus) kept under human care in a Zoo was found dead after 2 weeks presenting signals of weight loss and hyporexia. Histopathological revealed a necrotizing encephalitis. Although it was not observed microscopically, Sarcocystis sp infection was detected in brain tissue from molecular assays. These infections have been rarely described in neotropical primates, particularly associated with tissue lesions.

First reported human case of isolation of Vagococcus fluvialis from the urine of a former zoo clerk in Japan: a case report.

BACKGROUND: Vagococcal infections are extremely rare in humans. There are limited studies on the optimal methods for identification, antimicrobial susceptibility testing, and clinical manifestations of vagococcal infections. Herein, we report a patient with a urinary tract infection who had Vagococcus fluvialis in the urine. CASE PRESENTATION: An 84-year-old man presented to our urology department with a fever that had persisted for several days. He previously worked as a zoo clerk. The patient underwent a left nephroureterectomy for ureteral cancer 5 years ago, and total cystectomy and right cutaneous ureterostomy for muscle-invasive bladder cancer 1 year prior. He was empirically treated with 500 mg of levofloxacin intravenously every 24 h for the urinary tract infection. V. fluvialis was detected in his urine samples and Pseudomonas aeruginosa was detected in his urine and blood samples. Two bacterial species were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. He was administered intravenous levofloxacin for approximately 1 week, followed by oral levofloxacin for another week, after which the infections were eradicated. CONCLUSIONS: To the best of our knowledge, this is the first report of V. fluvialis detected in human urine in Japan. Vagococcus spp. is commonly isolated from fish or animals, and based on the patient's work history, it is possible that the patient was a carrier because of transmission from animals.

Mortality associated with SARS-CoV-2 in nondomestic felids.

Between September and November 2021, 5 snow leopards (Panthera uncia) and 1 lion (Panthera leo) were naturally infected with severe acute respiratory coronavirus 2 (SARS-CoV-2) and developed progressive respiratory disease that resulted in death. Severe acute respiratory syndrome coronavirus 2 sequencing identified the delta variant in all cases sequenced, which was the predominant human variant at that time. The time between initial clinical signs and death ranged from 3 to 45 days. Gross lesions in all 6 cats included nasal turbinate hyperemia with purulent discharge and marked pulmonary edema. Ulcerative tracheitis and bronchitis were noted in 4 cases. Histologically, there was necrotizing and ulcerative rhinotracheitis and bronchitis with fibrinocellular exudates and fibrinosuppurative to pyogranulomatous bronchopneumonia. The 4 cats that survived longer than 8 days had fungal abscesses. Concurrent bacteria were noted in 4 cases, including those with more acute disease courses. Severe acute respiratory syndrome coronavirus 2 was detected by in situ hybridization using probes against SARS-CoV-2 spike and nucleocapsid genes and by immunohistochemistry. Viral nucleic acid and protein were variably localized to mucosal and glandular epithelial cells, pneumocytes, macrophages, and fibrinocellular debris. Based on established criteria, SARS-CoV-2 was considered a contributing cause of death in all 6 cats. While mild clinical infections are more common, these findings suggest that some SARS-CoV-2 variants may cause more severe disease and that snow leopards may be more severely affected than other felids.

Identification of Haemoproteus infection in an imported grey crowned crane (Balearica regulorum) in China.

Blood parasites from the order Haemosporida infect many vertebrates and cause malaria-like diseases. In this study, a haemosporidian infection was detected in a sick grey crowned crane imported into China using a combination of morphological and molecular approaches. Blood samples were collected from the jugular vein and processed for morphological identification of infective parasites using stained blood smears and microscopy. No merogony occurs in the blood cells, and sporadic pigment granules were observed. Nested-PCR assays were employed for a molecular examination, which indicated that the cytb gene of this parasite had 94.1-94.9% identity to Haemoproteus antigonis. Subsequently, its mitochondrial genome structure was determined by high-throughput sequencing using the DNBSEQ-T7 platform. The determined structure was confirmed by the Sanger sequencing using amplicons. The mitochondrial genome obtained for this parasite exhibited a low CG content (32.0%) and possessed three protein-coding genes, encoding 1068 amino acids, which constituted 53.7% of the genome. Phylogenetic analysis indicated that this parasite clustered with Haemoproteus sp. is detected in grey crowned cranes from Africa. This parasite was likely acquired during importation of this animal; thus, strict quarantine of imported ornamental animals is required to prevent the entry of new pathogens.

Safety and immunogenicity of an adjuvanted recombinant spike protein-based severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine, SpikeVet™, in selected Carnivora, Primates and Artiodactyla in Australian zoos.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can infect a broad range of animal species and has been associated with severe disease in some taxa. Few studies have evaluated optimal strategies to mitigate the risk to susceptible zoo animals. This study evaluated the safety and immunogenicity of a protein-based veterinary SARS-CoV-2 vaccine (SpikeVet™) in zoo animals. Two to three doses of SpikeVet™ were administered intramuscularly or subcutaneously 3-4 weeks apart to 354 zoo animals representing 38 species. SpikeVet™ was very well tolerated across all species. Minor adverse effects were observed in 1.69% of animals vaccinated, or 1.04% of vaccine doses administered. Preliminary immunogenicity analyses in representative carnivores (meerkats, lions) and an artiodactylid (domestic goat) showed SpikeVet™-immunized animals developed serum antibodies able to neutralize a range of SARS-CoV-2 variants, including the vaccine-homologous Wuhan and Mu variants, as well as vaccine-heterologous Omicron BA.2 and XBB.1 strains. Prior to vaccination, all eight lions were seropositive for Wuhan strain by surrogate viral neutralization testing, suggesting past infection with SARS-CoV-2 or cross-reactive antibodies generated by another closely related coronavirus. These results from a range of zoo species support the ongoing development of SpikeVet™ as a safe and effective veterinary SARS-CoV-2 vaccine.

Case study: Visual barriers reduce pacing in captive tigers.

Captive large felines are prone to abnormal repetitive behaviors like pacing, which are associated with welfare issues. Visual contact without the opportunity to engage in appropriate behavior is known to increase pacing. To better understand the relationship between pacing and conspecific visual contact, we investigated this effect by conducting a barrier experiment on a male-female pair of Sumatran tigers (Panthera tigris sumatrae) in Rotterdam Zoo, the Netherlands. The tigers were exposed to four consecutive housing treatments: (i) housed in the same enclosure (baseline), (ii) housed in separate enclosures with visual contact, (iii) housed in separate enclosures without visual contact, and (iv) housed in the same enclosure after the separation. We used focal and scan sampling to measure pacing and recorded the number of visitors. Moreover, we applied scan sampling to measure activity. Overall, our results indicate that the tigers paced significantly more when housed in separate enclosures with conspecific visual contact. Moreover, our results suggest that limiting visual contact between neighboring tigers can mitigate pacing. Implementing these findings in tiger husbandry and enclosure design has the potential to improve animal welfare zoo populations of large felines.

Elephant Gestation: Insights Into Idiopathic Abortions and Stillbirth.

The declining African and Asian elephant populations emphasize the importance of a backup population. Successful reproduction in captivity plays a key role in maintaining such a genetically diverse ex situ population but is challenged by reproductive loss in the form of abortions and stillbirths. The elephants' biphasic prolactin pattern led us to predict a higher incidence of abortions during the time of reduced prolactin concentrations. Therefore, this study focuses on the identification of months during elephant gestation which are prone to loss of pregnancy. A metric was developed to identify the fetal age of aborted calves based on the fetal mass using a regression model. Data on idiopathic abortions in captive and wild elephants collected from zoos, tourist camps, semi-captive, and free-ranging populations since 1974 were analyzed, revealing a significantly higher prevalence of abortions during the 15th and 17th month of gestation. Additionally, the prevalence of stillbirths in the 22nd month of gestation between 2000 and 2023 was assessed. Although stillbirths showed a declining trend over time, the average prevalence between 2019 and 2023 was still 2.8%. Consequently, the 15th, 17th, and 22nd month of gestation were identified as stages prone to pregnancy loss. These findings underscore the necessity of researching risk factors and preventative measures for pregnancy loss in these 3 months, especially exploring a possible link with prolactin during the 15th and 17th month of gestation. The identification of stages prone to fetal loss is a key step towards enhancing elephant reproductive success and welfare.

The effects of anthropogenic noise on frogs housed on exhibit at a public aquarium.

Anuran behavior and reproduction are dominated by vocalizations, rendering them vulnerable to the effects of signal masking. For anurans on display in zoos and aquaria, a major source of ambient noise is visitors, which pose a unique source of potential anthropogenic signal masking. Call characteristics (total call duration, and minimum and maximum call frequencies) of three populations of dendrobatids (Dendrobates leucomelas, Epipedobates tricolor, and Ranitomeya imitator) on public display were investigated at time periods of increasing visitor-related noise (closed, off-peak, and peak aquarium visiting hours) to determine if there were changes in call characteristics that correlated with changes in visitor noise levels. The data revealed that call length increased with more visitor noise for D. leucomelas and E. tricolor, with their longest calls during peak hours, and all three species had their shortest calls during closed hours. Both minimum and maximum call frequencies increased with more visitor noise for E. tricolor and R. imitator, with their highest frequencies during peak hours, and lowest frequencies during closed hours. This study found evidence that anurans on public display adjust their vocalizations in the presence of visitor noise. These findings support expanded monitoring of ambient noise for animals on public display to determine if noise poses significant effects that might influence well-being or reproduction.

Never be mute about bird welfare: Swanning around with environmental enrichment.

Environmental enrichment (EE) is commonly provided to animals managed under human care, being beneficial to behavioral diversity and improving animal welfare. Use of EE appears to be particularly beneficial to individual wild animals spending a short period of time in captivity, for example, as part of conservation or rehabilitation programs. This paper documents a case study on the application and relevance of EE for a group of captive mute swans housed in a rescue center. Observational data were analyzed for two groups of juvenile swans that were provided with a physical EE device to increase time spent foraging. Periods of no EE were observed and compared to data from when birds were provided with EE. Results show that EE promoted foraging time and helped to reduce long periods of inactivity in captive birds. EE helped to reduce occurrence of captive-focused (i.e., abnormal behaviors) although these was already seen at very low rates. Inactivity as a measure of welfare in captive swans specifically (and waterbirds generally) should be further investigated to understand potential impacts on bird health. Our research shows the benefits of simple and easy-to-use EE devices on captive animal behavior and how use of EE for individuals spending a short amount of time in captivity (e.g., within a rescue center) could ensure diversity of behavior patterns and promote the performance of adaptive behaviors upon release to the wild.

Comparing the effectiveness of flagship species in zoo interpretation videos involving dialogic-based narrative approaches.

Zoological institutions frequently engage in indirect conservation activities as global conservation targets suggest a need for raising public awareness and engagement in biodiversity conservation. However, research suggests that while members of the public are typically aware of general conservation issues, they are often uncertain of simple and practical actions they take that will be impactful. In light of current conservation goals and targets, and the need for social science research to address the environmental learning and behavior change gap, this study builds upon prior action-based environmental education research and tests the efficacy of ex situ environmental education in supporting in situ conservation. Zoos typically employ flagship species to center their conservation messaging due to the purported draw of charismatic species. Using outreach videos with a dialogic-based narrative approach, we evaluated the efficacy of different flagships for conservation, comparing both species-focused versus generic conservation messaging and charismatic versus less charismatic species ambassadors. We found that zoo conservation outreach videos using dialogic-based narrative approaches were equally impactful regardless of the level of focus (i.e., species vs. broader biodiversity) or charisma level of the focal taxa.

Assessing the impact of environmental enrichment on behavior in understudied armadillo species: A case study.

The implementation of environmental enrichment (EE) can be effective in promoting zoo animal welfare by enhancing the performance of natural or species-typical behaviors. Research on the effects of EE is biased towards larger mammalian species, with less charismatic species being overlooked. Armadillos are one such overlooked example. A captive environment that results in inactivity, obesity, and associated poor health can negatively affect armadillo well-being. The aim of this study was to evaluate how the implementation of four physical object-based EEs could positively affect the behaviors of three armadillo species, housed in four similar exhibits. Behavioral data were collected both before (baseline) and during (treatment) EE periods, alongside of visitor number and environmental temperatures. The EE comprised of a plastic ball or a cardboard tube or a cardboard box, or a scatter-feed, and these were rotated each week of study until each exhibit had received them in turn. Despite the presence of different EE types, activity remained low throughout the study. However, results suggest that the plastic ball and cardboard box increased exploratory behaviors in the armadillos, but no overall increase in activity was noted during the scatter feed. Visitor presence had no effect on armadillo activity, and armadillos showed reduced activity with increasing environmental temperature. Overall, the use of physical object-based EE promoted beneficial natural behaviors in zoo-housed armadillos, but environmental conditions (i.e., temperature) also impacted armadillo activity, suggesting a complicated relationship between an enclosure's environmental variable and any behavioral husbandry measures.

Moving Toward Evidence-Based Care: Measuring Preferences in Zoo-Housed Aquatic Turtles Using 24-h Remote Monitoring.

Providing evidence-based care to animals is a common goal in zoos and aquariums. However, there are many understudied species with a lack of information available, which may result in suboptimal care for individuals of these species. Using preference tests to ask individual animals what they want can be an effective means to enhance their care, and subsequently, their welfare. Using 24-h remote monitoring, we assessed substrate, enclosure feature, and lighting preferences of five aquatic turtles (two wood turtles, one Blanding's turtle, and two spotted turtles) that share an indoor enclosure at Lincoln Park Zoo. In addition, we compared enclosure feature and lighting preferences between winter and summer to determine whether the turtles had changing seasonal preferences despite being housed indoors. We found that the wood turtles and Blanding's turtle had substrate preferences, whereas the spotted turtles did not. In addition, we found that the turtles' enclosure feature preferences were consistent between winter and summer, but the amount of time spent on land, logs, and in water varied by individual. Finally, we found that the turtles utilized areas with ambient lighting more than areas with a heat lamp or UV lamp; this was consistent between winter and summer. The turtle preferences observed in this study can contribute to the development of care strategies that support their welfare and, in turn, may inspire similar preference studies at other institutions as we collectively advance toward providing animals of all taxonomic groups with evidence-based care.

Visitor engagement with zoo staff and interpretive demonstrations increases visitor stay time in two free-flight aviaries.

Understanding how visitors spend time in zoo exhibits is important as zoological facilities work to enhance visitor experience and conservation education outcomes. We explored a variety of factors we hypothesized would influence visitor stay times in two aviaries at Disney's Animal Kingdom®. Within these aviaries, visitors could utilize educational materials, listen to staff-led demonstrations, or talks, view animal training sessions, or speak directly with animal care or education staff. We observed visitors and recorded the opportunities they utilized in the exhibit. Visitors to either aviary who conversed with exhibit-specific staff stayed between 67% and 89% longer than visitors who did not. In addition, visitors who used a bird guide while in either aviary stayed between 59% and 82% longer than those who did not. Those who listened to an animal care talk or engaged with a staff-led demonstration stayed between 50% and 68% longer than those who did not have the option to participate in such activities. This study provides insight into how exhibit offerings can influence visitor behavior. Implementing strategies used in this study to increase visitors' time in an exhibit may enhance visitor experience and improve learning outcomes.

Ancestral chromosomal signatures of Paenungulata (Afroteria) reveal the karyotype of Amazonian manatee (Trichechus inunguis, Sirenia: Trichechidae) as the oldest among American manatees.

BACKGROUND: Chromosomal painting in manatees has clarified questions about the rapid evolution of sirenians within the Paenungulata clade. Further cytogenetic studies in Afrotherian species may provide information about their evolutionary dynamics, revealing important insights into the ancestral karyotype in the clade representatives. The karyotype of Trichechus inunguis (TIN, Amazonian manatee) was investigated by chromosome painting, using probes from Trichechus manatus latirostris (TML, Florida manatee) to analyze the homeologies between these sirenians. RESULTS: A high similarity was found between these species, with 31 homologous segments in TIN, nineteen of which are whole autosomes, besides the X and Y sex chromosomes. Four chromosomes from TML (4, 6, 8, and 9) resulted in two hybridization signals, totaling eight acrocentrics in the TIN karyotype. This study confirmed in TIN the chromosomal associations of Homo sapiens (HSA) shared in Afrotheria, such as the 5/21 synteny, and in the Paenungulata clade with the syntenies HSA 2/3, 8/22, and 18/19, in addition to the absence of HSA 4/8 common in eutherian ancestral karyotype (EAK). CONCLUSIONS: TIN shares more conserved chromosomal signals with the Paenungulata Ancestral Karyotype (APK, 2n = 58) than Procavia capensis (Hyracoidea), Loxodonta africana (Proboscidea) and TML (Sirenia), where TML presents less conserved signals with APK, demonstrating that its karyotype is the most derived among the representatives of Paenungulata. The chromosomal changes that evolved from APK to the T. manatus and T. inunguis karyotypes (7 and 4 changes, respectively) are more substantial within the Trichechus genus compared to other paenungulates. Among these species, T. inunguis presents conserved traits of APK in the American manatee genus. Consequently, the karyotype of T. manatus is more derived than that of T. inunguis.

Anthropogenic Transmission of SARS-CoV-2 from Humans to Lions, Singapore, 2021.

In Singapore, 10 captive lions tested positive for SARS-CoV-2 by real-time PCR. Genomic analyses of nanopore sequencing confirmed human-to-animal transmission of the SARS-CoV-2 Delta variant. Viral genomes from the lions and zookeeper shared a unique spike protein substitution, S:A1016V. Widespread SARS-CoV-2 transmission among humans can increase the likelihood of anthroponosis.

Characterization of a Tigecycline-Resistant and blaCTX-M-Bearing Klebsiella pneumoniae Strain from a Peacock in a Chinese Zoo.

In Chinese zoos, there are usually specially designed bird parks, similar to petting zoos, that allow children and adults to interact with diverse birds. However, such behaviors present a risk for the transmission of zoonotic pathogens. Recently, we isolated eight strains of Klebsiella pneumoniae and identified two blaCTX-M-positive strains from 110 birds, including parrots, peacocks, and ostriches, using anal or nasal swabs in a bird park of a zoo in China. There, K. pneumoniae LYS105A was obtained from a diseased peacock with chronic respiratory diseases by a nasal swab, which harbored the blaCTX-M-3 gene and exhibited resistance to amoxicillin, cefotaxime, gentamicin, oxytetracycline, doxycycline, tigecycline, florfenicol, and enrofloxacin. According to an analysis by whole-genome sequencing, K. pneumoniae LYS105A belongs to serotype ST859 (sequence type 859)-K19 (capsular serotype 19) and contains two plasmids, of which pLYS105A-2 can be transferred by electrotransformation and harbors numerous resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91. The above-mentioned genes are located in a novel mobile composite transposon, Tn7131, which makes horizontal transfer more flexible. Although no known genes were identified in the chromosome, a significant increase in SoxS upregulated the expression levels of phoPQ, acrEF-tolC, and oqxAB, which contributed to strain LYS105A acquiring resistance to tigecycline (MIC = 4 mg/L) and intermediate resistance to colistin (MIC = 2 mg/L). Altogether, our findings show that bird parks in zoos may act as important vehicles for the spread of multidrug-resistant bacteria from birds to humans and vice versa. IMPORTANCE A multidrug-resistant ST859-K19 K. pneumoniae strain, LYS105A, was obtained from a diseased peacock in a Chinese zoo. In addition, multiple resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91 were located in a novel composite transposon, Tn7131, of a mobile plasmid, implying that most of the resistance genes in strain LYS105A can be moved easily via horizontal gene transfer. Meanwhile, an increase in SoxS can further positively regulate the expression of phoPQ, acrEF-tolC, and oqxAB, which is the key factor for strain LYS105A to develop resistance to tigecycline and colistin. Taken together, these findings enrich our understanding of the horizontal cross-species spread of drug resistance genes, which will help us curb the development of bacterial resistance.

Chromosome evolution in Iberolacerta, a genus that deviates from the standard karyotype formula of Lacertidae.

This paper describes the preparation of flow-sorted chromosome paints from the Iberian Rock lizard Iberolacerta monticola, exemplifying their subsequent use in cross-species comparisons of chromosome painting. We carried out comparative analyses of chromosome evolution in the congeneric species I. galani and I. bonnali, as well as in two other species of Lacertini (Lacerta schreiberi and Timon lepidus) whose sex chromosomes were also studied through comparative genomic hybridization. Most species of Lacertini possess a diplod number of 2n = 38, with 36 acrocentric macrochromosomes and 2 microchromosomes. However, the nine species included in the genus Iberolacerta do not possess microchromosomes. Furthermore, very conspicuous differences from the standard Lacertini karyotype were observed in the three Pyrenean species of this genus, which included several biarmed metacentrics and a Z1Z2W multiple sex-chromosome system. With the possible exception of L. schreiberi, all the species of the family Lacertidae described to date appear to share homologous Z chromosomes, which date back to the last common ancestor of the whole group. We provide conclusive evidence that L. schreiberi should no longer be considered an exception to this rule, and demonstrate that the loss of microchromosomes in Iberolacerta was produced by their fusion to a middle-sized chromosome. Furthermore, we show that the multiple sex-chromosome system of the Pyrenean species of Iberolacerta originated from the fusion of the ancestral W chromosome with one of the shortest autosomes, and provide additional evidence of the fast evolution of DNA sequences linked to the W chromosome in Lacertini.

Successful surgical correction of a cataract in a chimpanzee (Pan troglodytes) via field setting at a zoological facility.

A 41-year-old male vasectomized, zoo-housed chimpanzee (Pan troglodytes) presented with progressive visual deficits due to bilateral cataract formation. Phacoemulsification and lenticular implant were performed by a veterinary and human board-certified ophthalmologist team in a field setting. Post-operative healing occurred without complication, and the patient returned to the troop with improved vision.

Genomic resources for Asian (Elephas maximus) and African savannah elephant (Loxodonta africana) conservation and health research.

We provide novel genomic resources to help understand the genomic traits involved in elephant health and to aid conservation efforts. We sequence 11 elephant genomes (5 African savannah, 6 Asian) from North American zoos, including 9 de novo assemblies. We estimate elephant germline mutation rates and reconstruct demographic histories. Finally, we provide an in-solution capture assay to genotype Asian elephants. This assay is suitable for analyzing degraded museum and noninvasive samples, such as feces and hair. The elephant genomic resources we present here should allow for more detailed and uniform studies in the future to aid elephant conservation efforts and disease research.

Zoo studies in primate physiology, health, and welfare.

Researchers have been studying primates in zoos for more than half a century. There are numerous benefits to conducting research with zoo collections, such as access to a variety of species, ease of sample collection, and the potential to manipulate some research variables. While much of the primate research conducted in zoos is behavioral, there also is a tradition of research focused on reproduction and endocrinology, especially in North America. The contributions to this special issue exemplify how this tradition continues today through a collection of articles on basic and applied research on reproduction and using physiological measures of health and welfare that could be beneficial across primate taxa. As is the case for primatological research in zoos more broadly, the articles in this special issue reflect a taxonomic bias for great apes despite the high species diversity found across zoo collections. Given this bias as well as the threat of extinction faced by many species, there remains a pressing need to increase primatology in zoos through research dedicated to both conservation in the wild and wellbeing in human care.