The Effects of Enrichment on Zoo-Housed Scarlet Ibis Behavior.

Good zoo animal welfare is commonly promoted with environmental enrichment; however, some species are less likely to be offered enrichment than others. This study tested the effect of enrichment on a group of scarlet ibises from Zoomarine Algarve, Portugal. The study consisted of a first baseline condition, followed by four types of enrichment displays with individual presentations, a post-enrichment condition, and a post-enrichment with all enrichment types presented simultaneously. The enrichment types chosen were physical, with a tidal simulation in the lakes of the enclosure; nutritional, presenting mussels in a plastic mesh tube; sensory, by playing scarlet ibises calls; and tactile, with brushes through the habitat. The data collection was performed for 21 days between January and March 2021, 3 times a day, using scan sampling and instantaneous time sampling every 2 min. The comparison between conditions revealed that most behaviors showed similarity between the baseline and post-enrichment conditions, suggesting that after removing the enrichment, the behaviors returned to their initial baseline. It was also found that each enrichment influenced different types of behaviors and these behaviors also changed depending on the time of day. Exploratory behavior was only associated with the presence of enrichment, and vocalizations were only heard after the sensory enrichment was performed. This study demonstrates that the use of physical and tactile enrichments increased activity and decreased stationary behaviors in this group of ibises and may be used to improve their lives in zoological contexts if included in the housing and husbandry protocols.

The plasma phospholipidome of the bottlenose dolphin (Tursiops truncatus) is modulated by both sex and developmental stage.

Lipidomics represent a valid complementary tool to the biochemical analysis of plasma in humans. However, in cetaceans, these tools have been unexplored. Here, we evaluated how the plasma lipid composition of Tursiops truncatus is modulated by developmental stage and sex, aiming at a potential use of lipidomics in integrated strategies to monitor cetacean health. We characterized the fatty acid profile and detected a total of 26 fatty acids in T. truncatus plasma. The most abundant fatty acids were palmitic acid (C16:0), stearic acid (C18:0) and oleic acid (C18:1n-9). Interestingly, there are consistent differences between the fatty acid profile of mature female and mature male specimens. Phospholipidome analysis identified 320 different lipid species belonging to phosphatidylcholine (PC, 105 lipid species), lysophosphatidylcholine (42), phosphatidylethanolamine (PE, 67), lysophosphatidylethanolamine (18), phosphatidylglycerol (14), lysophosphatidylglycerol (8), phosphatidylinositol (14), lysophosphatidylinositol (2), phosphatidylserine (3), sphingomyelin (45) and ceramides (2) classes. The statistical analysis of the phospholipidome showed that its composition allows discriminating mature animals between sexes and mature males from immature males. Notably, discrimination between sexes is mainly determined by the contents of PE plasmalogens and lysophospholipids (LPC and LPE), while the differences between mature and immature male animals were mainly determined by the levels of PC lipids. This is the first time that a correlation between developmental stage and sex and the lipid composition of the plasma has been established in cetaceans. Being able to discern between age and sex-related changes is an encouraging step towards using these tools to also detect differences related to disease/dysfunction processes.

First Documentation of Exophiala spp. Isolation in Psittaciformes.

Several fungi species are reported to act as opportunistic agents of infection in avian species. After the isolation of Exophiala spp., a dematiaceous fungal pathogen associated with a mucosal lesion in a military macaw (Ara militar), samples were collected from another 24 birds of the order Psittaciformes to study the possibility of Exophiala spp. being part of the commensal microbiota of these animals or its possible association with other clinical conditions. Swab samples were collected from the trachea and/or choanae of the birds and inoculated in Sabouraud chloramphenicol agar for fungal isolation. After incubation, fungal species were identified through their macroscopic and microscopic morphology. The presence of Exophiala spp. was identified in 15 of the 25 birds sampled and no statistical association was found between the clinical record of the birds and the fungal isolation. Our results suggest that Exophiala spp. can colonize the upper respiratory airways of psittaciform birds and has a low pathogenic potential in these animals. To the authors' knowledge, this is the first report of Exophiala spp. isolation from samples of the upper respiratory tract of Psittaciformes.

Seroprevalence of Toxoplasma gondii in Pinnipeds under Human Care and in Wild Pinnipeds.

Toxoplasma gondii infection has been reported in numerous species of marine mammals, some of them with fatal consequences. A serosurvey for T. gondii infection was conducted in pinnipeds from an oceanographic park in Portugal (n = 60); stranded pinnipeds on the Portuguese coast (n = 10); and pinnipeds captured in Lorenzensplate, Germany (n = 99). Sera from 169 pinnipeds were tested for the presence of antibodies to T. gondii by the modified agglutination test with a cut-off titre of 25. An overall seroprevalence of 8.9% (95% confidence interval: 5.1-14.2) was observed. Antibody titres of 25, 50, 100, 1600 and ≥3200 were found in five (33.3%), two (13.3%), five (33.3%), one (6.7%) and two (13.3%) animals, respectively. Pinnipeds under human care had a seroprevalence of 20.0% (12/60), in contrast to 2.8% (3/109) in wild pinnipeds (p < 0.001). General results suggest a low exposure of wild pinnipeds to T. gondii, while the seroprevalence found in pinnipeds under human care highlights the importance of carrying out further studies. This is the first serological survey of T. gondii in pinnipeds in Portugal and the first infection report in South African fur seal (Arctocephalus pusillus pusillus).

The plasma phospholipidome of Tursiops truncatus: From physiological insight to the design of prospective tools for managed cetacean monitorization.

Plasma biochemical analysis remains one of the established ways of monitoring captive marine mammal health. More recently, complementary plasma lipidomic analysis has proven to be a valid tool in disease diagnosis and prevention, with the potential to validate and complement common biochemical analysis, providing a more integrative approach. In this study, we thoroughly characterized the plasma polar lipid content of Tursiops truncatus, the most common cetacean species held under human care. Our results showed that phosphatidylcholine, lysophosphatidylcholine, and sphingomyelins (CerPCho) are the most represented phospholipid classes in T. truncatus plasma. Palmitic, oleic, and stearic acids are the major fatty acid (FA) present esterified to the plasma polar lipids of this species, although some n-3 species are also remarkably present, namely eicosapentaenoic and docosahexaenoic acids. The polar lipidome identified by HILIC LC-MS allowed identifying 304 different lipid species. These species belong to the phosphatidylcholine (103 lipid species), lysophosphatidylcholine (35), phosphatidylethanolamine (71), lysophosphatidylethanolamine (20), phosphatidylglycerol (13), lysophosphatidylglycerol (5), phosphatidylinositol (15), lysophosphatidylinositol (3), phosphatidylserine (6) lysophosphatidylserine (1), and sphimgomyelin (32) classes. This was the first time that the dolphin plasma phospholipid profile was characterized, providing a knowledge that will be important to further understand lipid metabolism and physiological regulation in small cetaceans. Furthermore, this study proved the practicability of the use of plasma lipid profiling for health assessment in marine mammals under human care.

The use of posaconazole delayed-release tablets in the successful treatment of suspected mucormycosis in a bottlenose dolphin (Tursiops truncatus) calf.

Mucorales infections in cetaceans have a high mortality rate. This case report refers to a bottlenose dolphin calf with suspected mucormycosis treated with posaconazole. This antifungal agent was discontinued after 96 days of therapy, however, the infection relapsed. Posaconazole was then resumed for a total of 255 days, with no signs of disease reactivation. The retrospective analysis of posaconazole serum levels in this successful case showed concentrations varying between 5.18 and 11.63 mg/L.

Erysipelas vaccination protocols in dolphins Tursiops truncatus evaluated by antibody responses over twenty continuous years.

Erysipelas is an infection caused by Erysipelothrix rhusiopathiae that affects many different species around the world, including cetaceans. The acute septicemic form can rapidly cause death in bottlenose dolphins Tursiops truncatus. The ultimate goals of this long-term study were the development and identification of the most effective vaccination protocol against clinical erysipelas in T. truncatus using a commercially available swine vaccine, and to determine whether there is a need for a semi-annual vaccination versus an annual vaccination. The present study concentrated on the immunization of a dolphin population (7 wild-born and 22 captive-born individuals) with 2 swine vaccines, the European 'Eurovac Ery®' vaccine and the American 'ER Bac Plus®' vaccine, and immunological profile results over a 20-yr time period. The general protocol was a primo-vaccination (between 3 and 7 mo of age for calves) with or without a booster 1 mo post primo-vaccination and either annual or semi-annual vaccination thereafter. Sera were collected prior to vaccination, 2 wk post-vaccination and monthly. A dolphin-specific ELISA was developed to analyze the erysipelas-specific antibody response of vaccinated animals. The final ELISA results (n = 1362 samples from 29 animals at pre- and post-vaccination time) suggest that (1) there is a significant difference in antibody levels at the start of the vaccination between older and younger animals; (2) at least 3 vaccinations are necessary to obtain antibody levels above the levels at pre-vaccination; (3) thereafter, annual vaccinations seem sufficient to keep antibody levels above the levels at pre-vaccination; and (4) both vaccines induced similar responses. No case of erysipelas infection was observed in this population during the study.

Dirofilaria immitis in pinnipeds and a new host record.

BACKGROUND: Dirofilaria immitis is a mosquito-borne pathogen that is spreading worldwide, and the associated infection (i.e. dirofilariosis) is becoming a threat to animals and humans living in endemic areas. Little is known about the occurrence and risk of infection of D. immitis in pinnipeds. Here we report dirofilariosis by D. immitis in several pinniped species kept in captivity in Portugal. METHODS: Animals were housed in an oceanographic park located in Algarve, southern Portugal, a geographical area endemic for canine dirofilariosis. To assess the occurrence of D. immitis, blood was collected from the park's resident pinniped population, which consisted of 16 animals (5 common seals Phoca vitulina, 2 grey seals Halichoerus grypus, 3 California sea lions Zalophus californianus and 6 South African fur seals Arctocephalus pusillus pusillus). Dirofilaria immitis nematodes were detected by real-time PCR and by the presence of circulating antigens. In addition, modified Knott's technique was performed to detect circulating microfilariae. Necropsies and histopathological examination of two animals which died during the study were also conducted. RESULTS: Out of the 16 pinnipeds housed at the park, seven (43.8%) were positive for D. immitis by real-time PCR (3 P. vitulina, 2 Z. californianus and 2 A. p. pusillus), two of which (P. vitulina) were also positive for the nematode's antigen. Additionally, D. immitis microfilariae were detected in one A. p. pusillus. Furthermore, several D. immitis specimens were retrieved from the right ventricle and pulmonary arteries at the necropsy of one P. vitulina and one A. p. pusillus. CONCLUSIONS: This study provides new epidemiological data on D. immitis infection in pinnipeds diagnosed through clinical, molecular and pathological findings. Additionally, the South African fur seal is herein reported as a new host for this zoonotic filarioid. The situation herein described could also occur in other parks located in areas where canine dirofilariosis is endemic. Active surveillance and preventive measures of dirofilariosis in pinnipeds on a local and global scale are therefore vital to improve the early diagnosis and control of dirofilariosis.