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cies.The clouded leopard (Neofelis nebulosa) is classified as vulnerable on the International Union for the Conservation of Nature Red List of Threatened Species. However, diseases affecting this species across zoo populations are not well documented. The primary objective of this retrospective study was to identify common and significant causes of morbidity and mortality in captive-bred clouded leopards from European, Asian, and Australian institutions. Medical records from 44 zoological parks that held 271 clouded leopards from 1934 to 2017 were reviewed. Major causes of mortality in the dead leopards (n = 141) were respiratory disease (17%), maternal neglect and starvation (12%), generalized infectious disease (10%), digestive disease (10%), and trauma (10%). Six animals lived more than 20 yr and two were older than 22 yr. Diseases were recorded 344 times (average of two per leopard) in 166 living leopards. The body systems most frequently affected by disease in these 166 individuals were, in order of frequencyoo populations.Vector-borne Plasmodium spp. infect a wide range of bird species. Although infections may be asymptomatic, certain genera, especially those that evolved in regions without endemic malaria, appear particularly susceptible to symptomatic disease, leading to morbidity and mortality. High mortalities associated with malaria infections have been documented in captive species of Sphenisciformes, Somateria, and Larosterna, all genera that evolved in climates with low mosquito exposure. To better characterize trends in Plasmodium-related mortality in a zoological collection in New York, necropsy reports for birds of all three genera that died between 1998 and February 2018 were analyzed; comparisons were made between birds that died with or without evidence of malaria infection. A seasonal peak in deaths was observed in birds regardless of their malaria status. There was no significant difference in the age of birds at death between malaria-positive and malaria-negative animals. These results suggest that age and season of death were not associated with malaria status. To investigate an association between parasite lineage and clinical outcome, polymerase chain reaction was used to identify parasite lineage in necropsied birds as well as healthy birds sampled as part of surveillance studies. Twelve different Plasmodium lineages were identified. The relative prevalence of parasite lineages was compared between necropsy and surveillance samples. A single parasite lineage, SGS1 (species Plasmodium relictum), was significantly more likely to be found in surveillance samples; it was detected in a plurality of surveillance data but found in only one necropsy case. Other parasite lineages were more likely to be found in necropsies than in surveillance samples, most notably SEIAUR01 (species Plasmodium cathemerium). These data may be consistent with a difference in virulence between parasite lineages. This investigation has implications for the monitoring and care of vulnerable avian species.There are limited reports of the genetic characterization of Toxoplasma gondii infecting captive macropods in North America. A novel genotype, ToxoDB PCR-RFLP genotype 263, was reported from six wallabies at a zoological facility in Virginia, USA, prompting an investigation into the genotypes from T. gondii strains infecting macropods at a zoological park in Florida, USA. Cardiac muscle and/or lung samples from an agile wallaby (Macropus agilis, n = 1), red kangaroos (Macropus rufus, n = 8), red-necked wallaby (Macropus rufogriseus, n = 1), and a tammar wallaby (Macropus eugenii, n = 1) that died between 2014 and 2018 were collected. All 11 cases were confirmed to have died from systemic toxoplasmosis by histopathology and immunohistochemical staining. Multilocus PCR-RFLP genotyping of T. gondii was performed directly on tissue samples or on parasites isolated from myocardium by mouse bioassay. Two cases of toxoplasmosis were identified as the reported novel genotype, ToxoDB PCR-RFLP genotype 263, but no common source of exposure could be identified. Five cases were identified as genotype 2 (type III strain, haplogroup 3), and four cases were identified as genotype 216, which has been previously reported in North American wildlife. There were no overt differences in lesion severity or distribution related to genotype. These results suggest that the premise was contaminated with at least three genotypes of T. gondii causing systemic toxoplasmosis in macropods. The largest cluster of fatal toxoplasmosis in macropods in the study period occurred following severe rainfall flooding of the exhibit, suggesting the transmission of T. gondii by water and pointing out the importance of this transmission mechanism. In summary, our study revealed three T. gondii outbreaks that caused significant loss of macropods within 5 yr in a zoological facility in Florida. More studies are needed to understand transmission and prevention of toxoplasmosis in sensitive zoo animals.Agarose gel electrophoresis (AGE) has been widely implemented throughout veterinary medicine and for analysis of plasma proteins of avian and reptile species. Capillary zone electrophoresis (CZE) is becoming a standard method in human clinical pathology laboratories but has not widely been used for the analysis of animal samples. The objective of the present study was to compare protein fractions derived from AGE and CZE methods using plasma from the green turtle (Chelonia mydas). CAY10444 S1P Receptor antagonist Plasma samples were analyzed by AGE and CZE per manufacturer guidelines. The methods were assessed by CV analysis, Spearman's correlation, Passing-Bablok regression, and Bland Altman plots. CZE consistently resolved more fractions than AGE with three fractions observed in the prealbumin migrating region versus one for AGE and two fractions in the γ globulin region versus one for AGE. Compared with AGE, CZE showed a lower CV in intra-assay tests (1.0-4.9% vs 2.0-28.3%) and a lower or overlapping CV in interassay tests (1.0-10.6 vs 2.3-22.0). The prealbumin, α2 globulin, and β globulin fractions correlated the least between the methods (for all three fractions rs ≤ 0.28, P > 0.21). Moderate, significant correlations between AGE and CZE methods were observed for albumin (rs = 0.78, P less then 0.0001) and γ globulins (rs = 0.78, P less then 0.0001). CZE has a higher precision and ease of use over AGE and offers the opportunity to resolve additional protein fractions. This will necessitate the development of new conventions in placement of fraction delimits, definition of species-specific reference intervals, and evaluation of clinical utility in abnormal turtles.
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