Development and validation of a versatile non-invasive urinary steroidomics method for wildlife biomonitoring.

Wildlife conservation is often challenged by a lack of knowledge about the reproduction biology and adaptability of endangered species. Although monitoring steroids and related molecules can increase this knowledge, the applicability of current techniques (e.g. immunoassays) is hampered by species-specific steroid metabolism and the requisite to avoid invasive sampling. This study presents a validated steroidomics method for the (un)targeted screening of a wide range of sex and stress steroids and related molecules in urine using ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). In total, 50 steroids (conjugated and non-conjugated androgens, estrogens, progestogens and glucocorticoids) and 6 prostaglandins could be uniquely detected. A total of 45 out of 56 compounds demonstrated a detection limit below 0.01 ng µL-1. Excellent linearity (R2 > 0.99), precision (CV < 20 %), and recovery (80-120 %) were observed for 46, 41, and 39 compounds, respectively. Untargeted screening of pooled giant panda and human samples yielded 9691 and 8366 features with CV < 30 %, from which 84.1 % and 83.0 %, respectively, also demonstrated excellent linearity (R2 > 0.90). The biological validity of the method was investigated on male and female giant panda urine (n = 20), as well as pooled human samples (n = 10). A total of 24 different steroids were detected with clear qualitative and quantitative differences between human and giant panda samples. Furthermore, expected differences were revealed between female giant panda samples from different reproductive phases. In contrast to traditional biomonitoring techniques, the developed steroidomics method was able to screen a wide range of compounds and provide information on the putative identities of metabolites potentially important for reproductive monitoring in giant pandas. These results illustrate the advancements steroidomics brings to the field of wildlife biomonitoring in the pursuit to better understand the biology of endangered species.

Rapid facial mimicry in Platyrrhini: Play face replication in spider monkeys (Ateles fusciceps, Ateles hybridus, and Ateles paniscus).

Rapid facial mimicry (RFM), the rapid and automatic replication of facial expression perceived, is considered a basic form of empathy and was investigated mainly during play. RFM occurs in Catarrhini (Old World primates), but it is not still demonstrated in Platyrrhini (New World primates). For this reason, we collected video data on playful interactions (Nplay_interactions = 149) in three species of spider monkeys (Ateles fusciceps-N = 11, Ateles hybridus-N = 14, and Ateles paniscus-N = 6) housed at La Vallée des Singes and the ZooParc de Beauval (France). For the first time, we demonstrated the occurrence of RFM in Platyrrhini (analyzing 175 events). Players' sex, age, species, relationship quality, and kinship did not modulate RFM probably due to the species' complex fission-fusion dynamics and flexible interindividual social relationships. Compared to the absence of any playful expressions or the presence of only not replicated play face, RFM prolonged the session duration and was sequentially associated with more types of more intense offensive playful patterns (patterns aimed at attacking/pursuing the playmate). We proposed that RFM may favor synchronization and context sharing between players, thus decreasing the risk of behavior misinterpretation while simultaneously fostering a more competitive nature of play. In conclusion, this study stimulates additional research on the evolutionary origins of motor mimicry in primates, possibly dating back to before the divergence of New and Old World monkeys. Furthermore, it also points toward the possibility that RFM may not always lead to cooperation but also to competition, depending on the context and species' social and cognitive features.

Visual and acoustic exploratory behaviors toward novel stimuli in Antillean manatees (Trichechus manatus manatus) under human care.

Exploratory behaviors describe the actions performed by an animal to obtain information on an object, environment, or individual by using its different senses. Exploration is described in some marine mammals, but not yet in manatees. Our study investigated behavioral and acoustic responses of two groups of Antillean manatees (N = 12 and N = 4) housed in zoological parks toward various stimuli involving three sensory modalities: visual, tactile, and auditory. Simultaneous audio and video recordings were collected during three periods of time (i.e., before, during, and after the presentation of all stimuli). Behaviors related to interest, social behaviors, the number and type of calls produced, and their frequency and duration were recorded and analyzed. Manatees reacted more to submerged stimuli than to out-of-water and sound stimuli, with an increase in approach, social contacts, and number of vocalizations. The proportion of squeaks and squeals call types also varied according to stimuli, and call entropy and F0 range varied according to periods. Our results suggest that manatees display sensory preferences when exploring stimuli, with more interest in manipulable stimuli, supporting the importance of their somatic perception. We highlight the need for particular enrichment programs (i.e., involving submerged objects) in zoological facilities. By displaying social contacts and by producing vocalizations, manatees communicate information such as their motivational state. The increase in call rate, harsh calls, and entropy values could be valid indicators of heightened arousal. We encourage further studies to associate acoustic recordings with ethological data collection to increase the understanding of manatees' behaviors andperception. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Retrospective Study of Intertarsal Joint Inflammation in Avian Species From a Zoological Institution.

A retrospective cross-sectional study was performed to measure the prevalence of avian intertarsal inflammation over a 5-year period, identify risk factors, and discuss treatment options. The authors hypothesized that: 1) long-legged birds would be more affected, 2) participation in a bird show would be a significant risk factor, and 3) young animals would be more frequently affected. Thirty-five clinical cases from 9 avian orders were included in the study. Statistical analysis indicated that the orders Ciconiiformes (9/150; 6%, 95% confidence interval [CI]: 3.2-11), Gruiformes (4/132; 3%, 95% CI: 1.2-7.5), and Pelecaniformes (7/152; 2.8%, 95% CI: 1.4-5.6) were significantly more affected than other orders (P < 0.01). Similarly, long-legged birds (21/35) had 9.8 times greater chance (P < 0.001, 95% CI: 4.7-21) of developing the condition compared with other birds. Participation in a free-flight show (22/35) was a significant risk factor (P < 0.001; odds ratio: 7.0, 95% CI: 3.3-15). Mean age at onset of clinical signs was 5.7 years, and being < 2 years-of-age during the study period was not a significant predictor of disease (P = 0.054). The tibial cartilage, a fragile fibrocartilaginous structure, was frequently affected (34%, 12/35). Treatment protocols included anti-inflammatory drugs, analgesic drugs, or both (94%, 33/35), low-level laser therapy (54%, 19/35), joint immobilization (34%, 12/35), intra-articular corticoid injections (20%, 7/35), surgical stabilization (17%, 6/35), physiotherapy (9%, 3/35), intra-articular hyaluronic acid (6%, 2/35) or platelet-rich plasma (3%, 1/35) injections, and chiropractic care (3%, 1/35). Overall recovery rate was 49% (17/35), and the condition was associated with a poor prognosis in chronic cases.

The impact of the land-to-sea transition on evolutionary integration and modularity of the pinniped backbone.

In this study, we investigate how the terrestrial-aquatic transition influenced patterns of axial integration and modularity in response to the secondary adaptation to a marine lifestyle. We use 3D geometric morphometrics to quantify shape covariation among presacral vertebrae in pinnipeds (Carnivora; Pinnipedia) and to compare with patterns of axial integration and modularity in their close terrestrial relatives. Our results indicate that the vertebral column of pinnipeds has experienced a decrease in the strength of integration among all presacral vertebrae when compared to terrestrial carnivores (=fissipeds). However, separate integration analyses among the speciose Otariidae (i.e., sea lions and fur seals) and Phocidae (i.e., true seals) also suggests the presence of different axial organizations in these two groups of crown pinnipeds. While phocids present a set of integrated "thoracic" vertebrae, the presacral vertebrae of otariids are characterized by the absence of any set of vertebrae with high integration. We hypothesize that these differences could be linked to their specific modes of aquatic locomotion -i.e., pelvic vs pectoral oscillation. Our results provide evidence that the vertebral column of pinnipeds has been reorganized from the pattern observed in fissipeds but is more complex than a simple "homogenization" of the modular pattern of their close terrestrial relatives.

Conserved core microbiota in managed and free-ranging Loxodonta africana elephants.

The gut microbiota plays a crucial role in animal health and homeostasis, particularly in endangered species conservation. This study investigated the fecal microbiota composition of European captive-bred African savanna elephants (Loxodonta africana) housed in French zoos, and compared it with wild African savanna elephants. Fecal samples were collected and processed for DNA extraction and amplicon sequencing of the 16S rRNA gene. The analysis of α and ß diversity revealed significant effects of factors such as diet, daily activity, and institution on microbiota composition. Specifically, provision of branches as part of the diet positively impacted microbiota diversity. Comparative analyses demonstrated distinct differences between captive and wild elephant microbiomes, characterized by lower bacterial diversity and altered co-occurrence patterns in the captive population. Notably, specific taxa were differentially abundant in captive and wild elephants, suggesting the influence of the environment on microbiota composition. Furthermore, the study identified a core association network shared by both captive and wild elephants, emphasizing the importance of certain taxa in maintaining microbial interactions. These findings underscore the impact of environment and husbandry factors on elephant gut microbiota, highlighting the benefits of dietary enrichment strategies in zoos to promote microbiome diversity and health. The study contributes to the broader understanding of host-microbiota interactions and provides insights applicable to conservation medicine and captive animal management.

Captive Blue-and-yellow macaws (Ara ararauna) show facial indicators of positive affect when reunited with their caregiver.

In mammals, human-animal bonding is recognized as a source of positive affect for companion or farm animals. Because this remains unexplored in birds, we investigated captive parrots' perspective of the human-animal relationship. We used a classical separation-reunion paradigm and predicted that variations in parrots' facial displays and behaviours would indicate their appraisal of the relationship. The test was divided into three phases of two minutes each: the bird was placed in an unfamiliar environment with a familiar caregiver (union), then the bird was left alone (separation) and finally, the caregiver returned (reunion). The test was repeated 10 times for each bird and video recorded in order to analyze their behaviour. The data show significantly higher crown and nape feather heights, higher redness of the skin and higher frequency of contact-seeking behaviours during the union and reunion phases than during the separation phase during which they expressed long distance contact calls. We observed the expression of eye pinning during the union and reunion phases in one out of five macaws. We argue that variation in facial displays provides indicators of parrot's positive appraisal of the caretaker presence. Our results broaden the scope for further studies on parrots' expression of their subjective feelings.

Advancements in veterinary medicine: the use of Flowgy for nasal airflow simulation and surgical predictions in big felids (a case study in lions).

Flowgy is a semi-automated tool designed to simulate airflow across the nasal passage and detect airflow alterations in humans. In this study, we tested the use and accuracy of Flowgy in non-human vertebrates, using large felids as the study group. Understanding the dynamics of nasal airflow in large felids such as lions (Panthera leo) is crucial for their health and conservation. Therefore, we simulated airflow during inspiration through the nasal passage in three lions (Panthera leo), two of which were siblings (specimens ZPB_PL_002 and ZPB_PL_003), without breathing obstructions. However, one of the specimens (ZPB_PL_001) exhibited a slight obstruction in the nasal vestibule, which precluded the specimen from breathing efficiently. Computed tomography (CT) scans of each specimen were obtained to create detailed three-dimensional models of the nasal passage. These models were then imported into Flowgy to simulate the airflow dynamics. Virtual surgery was performed on ZPB_PL_001 to remove the obstruction and re-simulate the airflow. In parallel, we simulated the respiration of the two sibling specimens and performed an obstructive operation followed by an operation to remove the obstruction at the same level and under the same conditions as the original specimen (ZPB_PL_001). Thus, we obtained a pattern of precision for the operation by having two comparable replicas with the obstructed and operated specimens. The simulations revealed consistent airflow patterns in the healthy specimens, demonstrating the accuracy of Flowgy. The originally obstructed specimen and two artificially obstructed specimens showed a significant reduction in airflow through the right nostril, which was restored after virtual surgery. Postoperative simulation indicated an improvement of >100% in respiratory function. Additionally, the temperature and humidity profiles within the nostrils showed marked improvements after surgery. These findings underscore the potential of Flowgy in simulating nasal airflow and predicting the outcomes of surgical interventions in large felids. This could aid in the early detection of respiratory diseases and inform clinical decision-making, contributing to improved veterinary care and conservation efforts. However, further research is needed to validate these findings in other species and explore the potential of integrating Flowgy with other diagnostic and treatment tools in veterinary medicine.

Measuring the force of the tip of the elephants trunk.

Forces that animals can exert is of great interest in biology. Regarding the elephant trunk, the maximum mass that an elephant can lift with its trunk is known, but the pinching force of the trunk tip is unknown. We here present an original system to measure this force for an organ much coveted in soft robotics for example. It consists of:•A wooden box protecting the elephants and the measuring system with no protruding parts except the sensor. This box has an opening at the back to fill it with apples and a trap door at the front to release the apples.•Two load cells protruding from the box connected to an electronic system controlled by an Arduino card that records and sends the pinch force via Wi-Fi to a laptop while releasing a reward apple.•Depending on the threshold chosen, the elephant must pinch harder than the previous time to release the next apple. The repetition of this action allows us to approach the maximum force it can achieve.The system, tested on elephants at the Beauval Zooparc (France), has demonstrated that it is effective in measuring the pinching force of the tip of the trunk.

Aspergillosis in a colony of Humboldt penguins (Spheniscus humboldti) under managed care: a clinical and environmental investigation in a French zoological park.

Aspergillosis is pervasive in bird populations, especially those under human care. Its management can be critically impacted by exposure to high levels of conidia and by resistance to azole drugs. The fungal contamination in the environment of a Humboldt penguin (Spheniscus humboldti) group, housed in a French zoological park next to numerous large crop fields, was assessed through three serial sessions of surface sampling in nests, in 2018-20: all isolates were counted and characterized by sequencing. When identified as Aspergillus fumigatus, they were systematically screened for resistance mutations in the cyp51A gene and tested for minimal inhibitory concentrations (MICs) determination. At the same time, the clinical incidence of aspergillosis was evaluated in the penguin population by the means of systematic necropsy and mycological investigations. A microsatellite-based analysis tracked the circulation of A. fumigatus strains. Environmental investigations highlighted the substantial increase of the fungal load during the summer season (>12-fold vs. the other timepoints) and a large overrepresentation of species belonging to the Aspergillus section Fumigati, ranging from 22.7 to 94.6% relative prevalence. Only one cryptic species was detected (A. nishimurae), and one isolate exhibited G138S resistance mutation with elevated MICs. The overall incidence of aspergillosis was measured at ∼3.4% case-years, and mostly in juveniles. The analysis of microsatellite polymorphism revealed a high level of genetic diversity among A. fumigatus clinical isolates. In contrast, one environmental strain appeared largely overrepresented during the summer sampling session. In all, the rural location of the zoo did not influence the emergence of resistant strains.

Assessment of the Safety and Efficacy of an Oral Probiotic-Based Vaccine Against Aspergillus Infection in Captive-Bred Humboldt Penguins (Spheniscus humboldti).

Aspergillosis is a fungal infection caused mainly by Aspergillus fumigatus that often results in respiratory disease in birds. Aspergillosis is a major cause of morbidity and mortality in captive-bred penguin species. Currently, there is no registered vaccine to prevent aspergillosis. Recent research demonstrated that oral administration of gram-negative bacteria expressing high levels of galactose-α-1,3-galactose (α-Gal) modulates anti-α-Gal immunity and protects turkeys from clinical aspergillosis caused by experimental A. fumigatus infection. The role of anti-α-Gal immunity in penguins has not been studied. Here, we tested the distribution of α-1,3-galactosyltransferase (α1,3GT) genes in the fecal microbiome of Humboldt penguins (Spheniscus humboldti). The occurrence of natural anti-α-Gal antibodies (Abs) in sera and eggs of healthy Humboldt penguins was also assessed. A trial was then conducted to test whether oral administration of Escherichia coli Nissle, expressing high α-Gal levels, modulates anti-α-Gal immunity in a colony of Humboldt penguins. Animals in the vaccination and placebo groups were evaluated before the trial and followed for one year for aspergillosis detection using a diagnostic panel including computed tomography scans, capillary zone electrophoresis, 3-hydroxybutyrate levels, and anti-A. fumigatus Abs. Anti-α-Gal Abs were detected in sera (IgM and IgY) and eggs (IgY) of healthy penguins. Microbiota analysis and functional predictions revealed the presence of α1,3GT genes in the microbiota of Humboldt penguins and other penguin species. A strong decrease in anti-α-Gal IgM levels was observed in all animals in the placebo group three months after vaccination protocol. This decrease was not observed in E. coli Nissle-treated penguins. After the vaccination protocol, we found a positive correlation between anti-E. coli IgY and anti-α-Gal IgY in the E. coli Nissle group, suggesting a correlation between the presence of the bacteria and these Abs. During the study period, three penguins exhibited respiratory signs consistent with aspergillosis. Two were from the placebo group whose symptoms resolved with specific treatments, while a single vaccinated individual developed fatal respiratory aspergillosis eight months after the trial. We conclude that E. coli Nissle represents a safe potential probiotic with a protective effect against aspergillosis in Humboldt penguins that deserves to be further explored for therapeutic uses in these animals.

The relationship between distal trunk morphology and object grasping in the African savannah elephant (Loxodonta africana).

Background: During reach-to-grasp movements, the human hand is preshaped depending on the properties of the object. Preshaping may result from learning, morphology, or motor control variability and can confer a selective advantage on that individual or species. This preshaping ability is known in several mammals (i.e., primates, carnivores and rodents). However, apart from the tongue preshaping of lizards and chameleons, little is known about preshaping of other grasping appendages. In particular, the elephant trunk, a muscular hydrostat, has impressive grasping skills and thus is commonly called a hand. Data on elephant trunk grasping strategies are scarce, and nothing is known about whether elephants preshape their trunk tip according to the properties of their food. Methods: To determine the influence of food sizes and shapes on the form of the trunk tip, we investigated the morphology of the distal part of the trunk during grasping movements. The influence of food item form on trunk tip shape was quantified in six female African savannah elephants (Loxodonta africana). Three food item types were presented to the elephants (elongated, flat, and cubic), as well as three different sizes of cubic items. A total of 107 ± 10 grips per individual were video recorded, and the related trunk tip shapes were recorded with a 2D geometric morphometric approach. Results: Half of the individuals adjusted the shape of the distal part of their trunk according to the object type. Of the three elephants that did not preshape their trunk tip, one was blind and another was subadult. Discussion and perspectives: We found that elephants preshaped their trunk tip, similar to the preshaping of other species' hands or paws during reach-to-grasp movements. This preshaping may be influenced by visual feedback and individual learning. To confirm these results, this study could be replicated with a larger sample of elephants.

A novel male Japanese quail structural connectivity atlas using ultra-high field diffusion MRI at 11.7 T.

The structural connectivity of animal brains can be revealed using post-mortem diffusion-weighted magnetic resonance imaging (MRI). Despite the existence of several structural atlases of avian brains, few of them address the bird's structural connectivity. In this study, a novel atlas of the structural connectivity is proposed for the male Japanese quail (Coturnix japonica), aiming at investigating two lines divergent on their emotionality trait: the short tonic immobility (STI) and the long tonic immobility (LTI) lines. The STI line presents a low emotionality trait, while the LTI line expresses a high emotionality trait. 21 male Japanese quail brains from both lines were scanned post-mortem for this study, using a preclinical Bruker 11.7 T MRI scanner. Diffusion-weighted MRI was performed using a 3D segmented echo planar imaging (EPI) pulsed gradient spin-echo (PGSE) sequence with a 200 [Formula: see text]m isotropic resolution, 75 diffusion-encoding directions and a b-value fixed at 4500 s/mm2. Anatomical MRI was likewise performed using a 2D anatomical T2-weighted spin-echo (SE) sequence with a 150 [Formula: see text]m isotropic resolution. This very first anatomical connectivity atlas of the male Japanese quail reveals 34 labeled fiber tracts and the existence of structural differences between the connectivity patterns characterizing the two lines. Thus, the link between the male Japanese quail's connectivity and its underlying anatomical structures has reached a better understanding.

Social pressure drives "conversational rules" in great apes.

In the last decade, two hypotheses, one on the evolution of animal vocal communication in general and the other on the origins of human language, have gained ground. The first hypothesis argues that the complexity of communication co-evolved with the complexity of sociality. Species forming larger groups with complex social networks have more elaborate vocal repertoires. The second hypothesis posits that the core of communication is represented not only by what can be expressed by an isolated caller, but also by the way that vocal interactions are structured, language being above all a social act. Primitive forms of conversational rules based on a vocal turn-taking principle are thought to exist in primates. To support and bring together these hypotheses, more comparative studies of socially diverse species at different levels of the primate phylogeny are needed. However, the majority of available studies focus on monkeys, primates that are distant from the human lineage. Great apes represent excellent candidates for such comparative studies because of their phylogenetic proximity to humans and their varied social lives. We propose that studying vocal turn-taking in apes could address several major gaps regarding the social relevance of vocal turn-taking and the evolutionary trajectory of this behaviour among anthropoids. Indeed, how the social structure of a species may influence the vocal interaction patterns observed among group members remains an open question. We gathered data from the literature as well as original unpublished data (where absent in the literature) on four great ape species: chimpanzees Pan troglodytes, bonobos Pan paniscus, western lowland gorillas Gorilla gorilla gorilla and Bornean orang-utans Pongo pygmaeus. We found no clear-cut relationship between classical social complexity metrics (e.g. number of group members, interaction rates) and vocal complexity parameters (e.g. repertoire size, call rates). Nevertheless, the nature of the society (i.e. group composition, diversity and valence of social bonds) and the type of vocal interaction patterns (isolated calling, call overlap, turn-taking-based vocal exchanges) do appear to be related. Isolated calling is the main vocal pattern found in the species with the smallest social networks (orang-utan), while the other species show vocal interactions that are structured according to temporal rules. A high proportion of overlapping vocalisations is found in the most competitive species (chimpanzee), while vocal turn-taking predominates in more tolerant bonobos and gorillas. Also, preferentially interacting individuals and call types used to interact are not randomly distributed. Vocal overlap ('chorusing') and vocal exchange ('conversing') appear as possible social strategies used to advertise/strengthen social bonds. Our analyses highlight that: (i) vocal turn-taking is also observed in non-human great apes, revealing universal rules for conversing that may be deeply rooted in the primate lineage; (ii) vocal interaction patterns match the species' social lifestyle; (iii) although limited to four species here, adopting a targeted comparative approach could help to identify the multiple and subtle factors underlying social and vocal complexity. We believe that vocal interaction patterns form the basis of a promising field of investigation that may ultimately improve our understanding of the socially driven evolution of communication.

Analogous laterality in trunk movements in captive African elephants: A pilot study.

Lateralization of hand use in primates has been extensively studied in a variety of contexts, and starts to be investigated in other species and organs in order to understand the evolution of the laterality according to different tasks. In elephants, the orientation of the movements of the trunk has been observed mainly in feeding and social contexts, in free conditions. However, little is known about the influence of task complexity on trunk laterality. In this study, we compared the lateralization of the trunk in two conditions: standardized and free. We offered granules to six African elephants on each side of an opened trapdoor to create a constraining environment and reported the different behaviours employed and their orientation. In addition, we observed the same individuals in free conditions and noted the lateralization of the use of their trunk. We revealed a common right side preference in all our elephants, both in standardized and free conditions. This side bias was stronger in our constraining task, adding evidence for the task complexity theory. We finally described laterality in new behaviours in the literature on elephants, such as pinching, gathering or exploration with the trunk.

Use of a simplified non-invasive technic to monitor fecal progesterone metabolites and reproduction function in several zoo species: Efficacy of mini VIDAS® automate (bioMérieux).

Developing the zoos' ability to assess the reproductive status of the individuals they house is essential to improve the husbandry and management of these species. The use of non-invasive techniques such as fecal hormone analysis has been proven to be a simple and effective way to achieve this. Designed by bioMérieux, mini VIDAS® instrument is used in human and veterinary medicine to evaluate different endocrinological parameters, including serum or plasma progesterone. This study evaluates VIDAS® Progesterone (PRG) assay's efficacy to monitor fecal progestagens using a simple sample extraction protocol adapted to the zoo environment. We compared (1) VIDAS® PRG fecal profiles with established assays specifically designed for fecal progestagens analysis at the VetmedUni (Vienna, Austria) for okapis (Okapia johnstoni), greater one-horned rhinoceros (Rhinoceros unicornis), giraffes (Giraffa camelopardalis reticulata) and hippopotamus (Hippopotamus amphibius) (2) VIDAS® PRG fecal profiles with VIDAS® PRG serum profiles for African elephants (Loxodonta Africana), giant anteater (Myrmecophaga tridactyla) and white rhinoceros (Ceratotherium simum). Spearman mean correlations were: 0.6748 for African elephants (n = 2 animals), 0.7969 for giant anteater (n = 1 animal), 0.7926 for okapis (n = 2 animals), 0.6072 for greater one-horned rhinoceros (n = 4 animals), 0.6062 for giraffes (n = 4 animals) and 0.5740 for hippopotamus (n = 2 animals). Fecal progestagens analysis revealed estrous cycles in several species: 12.5 ± 0.5 weeks for African elephants (n = 2 cycles), 15.3 ± 1.1 days for okapis (n = 6 cycles), 44 ± 2.1 days for greater one-horned rhinoceros (n = 4 cycles) and 15.5 ± 0.5 days for giraffes (n = 4 cycles). We observed pregnancies in a giant anteater, an okapi and a hippopotamus. We observed a strong positive Spearman correlation (r > 0.60) for individuals exhibiting estrous cycles. These first results indicate that the mini VIDAS® can be used for monitoring of the reproductive status of non-domesticated species and can be a useful tool for the reproductive management through fecal progesterone analysis. A simple extraction protocol was suitable for sample preparation of fecal progesterone metabolite analysis. Further studies using a larger number of individuals per species at different reproductive stages could confirm the relevance of mini VIDAS® in the zoo community.

EQUID ALPHAHERPESVIRUS 9 OUTBREAK ASSOCIATED WITH MORTALITY IN A GROUP OF GREVY'S ZEBRA (EQUUS GREVYI) HOUSED IN A MIXED-SPECIES EXHIBIT.

A herd of seven captive-born Grevy's zebras (Equus grevyi) experienced an outbreak of nasal discharge and sneezing. Clinical signs, including lethargy and anorexia, were severe and acute in three animals, including a 16-mo-old male that died within 48 h. Treatment of two severely affected zebras included valacyclovir (40 mg/kg PO), meloxicam (0.6 mg/kg IM/PO), and cefquinome (2.5 mg/kg IM q48h). An adult female improved rapidly, and clinical signs resolved within 48 h of treatment. Administration of valacyclovir pellets was very complicated in a 2-mo-old female, and death occurred within 48 h. Histologic examination of the two individuals that died revealed severe fibrinonecrotic interstitial pneumonia with prominent hyaline membranes and type II pneumocyte hyperplasia. Additionally, the 16-mo-old male presented systemic endothelial activation with vascular thrombosis and necrosis and mild nonsuppurative meningoencephalitis. Herpesviral DNA was detected in the lungs of both individuals by nested polymerase chain reaction. The nucleic acid sequence of the amplicons showed 100% similarity with previously published equid alphaherpesvirus 9 sequences. Three additional animals developed mild nasal discharge only and recovered spontaneously. The zebras shared housing facilities with other species, including white rhinoceros (Ceratotherium simum), reticulated giraffe (Giraffa camelopardalis reticulata), and several antelope species. None of these animals showed clinical signs. Additionally, nasal swabs and whole blood samples were collected from cohoused white rhinoceroses (n = 3) and springboks (Antidorcas marsupialis, n = 3) as well as nasal swabs from cohoused reticulated giraffes (n = 4). Nucleic acid sequence from equid herpesviruses was not detected in any of these samples. The source of the infection in the zebras remains unclear.

Occurrence of Ten Protozoan Enteric Pathogens in Three Non-Human Primate Populations.

Non-human primate populations act as potential reservoirs for human pathogens, including viruses, bacteria and parasites, which can lead to zoonotic infections. Furthermore, intestinal microorganisms may be pathogenic organisms to both non-human primates and humans. It is, therefore, essential to study the prevalence of these infectious agents in captive and wild non-human primates. This study aimed at showing the prevalence of the most frequently encountered human enteric protozoa in non-human primate populations based on qPCR detection. The three populations studied were common chimpanzees (Pan troglodytes) in Senegal and gorillas (Gorilla gorilla) in the Republic of the Congo and in the Beauval Zoo (France). Blastocystis spp. were mainly found, with an occurrence close to 100%, followed by Balantidiumcoli (23.7%), Giardiaintestinalis (7.9%), Encephalitozoonintestinalis (1.3%) and Dientamoebafragilis (0.2%). None of the following protozoa were detected: Entamoebahistolytica, Enterocytozoonbieneusi, Cryptosporidiumparvum, C. hominis, Cyclosporacayetanensis or Cystoisosporabelli. As chimpanzees and gorillas are genetically close to humans, it is important to monitor them frequently against different pathogens to protect these endangered species and to assess potential zoonotic transmissions to humans.

Malignant Melanoma in a Long-legged Buzzard (Buteo rufinus): Characterization of Clinical and Histologic Lesions.

A 17-year-old, male long-legged buzzard (Buteo rufinus) presented for acute lethargy and black gelatinous tissue in both nostrils. Despite intensive care, the bird died within 2 days. A postmortem computed tomographic scan of the head revealed a complete obstruction of the rostral infraorbital sinus, later confirmed by necropsy. Postmortem examination also highlighted multifocal, black, infiltrating nodules of various sizes (2 mm to 2 cm in diameter) in almost all internal organs: lungs, heart (pericardium, myocardium, endocardium), kidneys, pancreas, adrenals, muscles, and bones (periosteum and medulla). Histologically, the nodules were composed of anaplastic cells containing abundant melanin pigment, consistent with a disseminated melanoma. Malignant melanomas are aggressive neoplasms frequently reported in mammals but rarely found in avian species. In Accipitridae and Falconidae, only 2 cases have been reported.

New insights in Cercopithecinae spermatozoa.

Sperm morphometric and morphologic data have been shown to represent useful tools for monitoring fertility, improving assisted reproduction techniques and conservation of genetic material as well as detecting inbreeding of endangered primates. We provide here for the first time sperm morphologic and morphometric data from Cercopithecus neglectus, Cercopithecus cephus, Papio papio and critically endangered Cercopithecus roloway, as well as comparative data from other Cercopithecinae species, i.e. Allochrocebus lhoesti, Mandrillus sphinx and Papio anubis. Following collection from the epididymis, spermatozoa were measured for each species for the following parameters: head length, head width, head perimeter, head area, midpiece length and total flagellum length, and the head volume, ellipticity, elongation, roughness and regularity were then calculated. Our data are consistent with both the general morphology and the morphometric proportions of Cercopithecinae sperm. Some specificities were observed, with C. cephus displaying a narrow head (width = 2.76 ± 0.26 µM) and C. roloway displaying a short midpiece (6.65 ± 0.61 µM). This data set represents an important contribution, especially for Cercopithecus roloway, one of the most endangered monkeys in the world, and further data on additional specimens coupled to data on mating systems and reproductive ecology should allow a better understanding of the mechanisms underlying these morphological differences across primate species.