Capillary Zone Electrophoresis in Humboldt Penguins (Spheniscus humboldti).

Electrophoresis is a useful diagnostic tool for detecting inflammation, including inflammation associated with infectious diseases (eg, aspergillosis in penguins). To our knowledge, reference intervals are not available for plasma proteins via electrophoresis in Humboldt penguins (Spheniscus humboldti). Therefore, preliminary reference intervals for blood plasma proteins measured by capillary zone electrophoresis were calculated for Humboldt penguins from a single zoological collection, and possible differences between the sexes and the ages of the birds were evaluated. Lithium heparinized plasma samples from 39 Humboldt penguins were analyzed. The following sex- and age-independent reference intervals were calculated: total protein 33.8-70.4 g/L, prealbumin 1.9-4.9 g/L, albumin 12.9-31.1 g/L, albumin: globulin ratio 0.7-1.7, α-globulins 4.5-11.6 g/L, ß-globulins 5.6-20.6 g/L, and γ-globulins 2.6-8.4 g/L. Male penguins had a significantly (P = 0.047) higher albumin: globulin ratio and lower percentage of ß-globulins (P = 0.015) in comparison with female penguins. Prealbumin (g/L) significantly (P = 0.021) decreased with increased age of the penguins. These results showed some differences between the sexes and ages of the penguins, which should be considered when interpreting the results. Further studies are needed to determine whether differences in other age groups or seasons exist, and also to evaluate which infectious diseases affect plasma proteins and how the reference values calculated here may deviate in ill penguins.

Clinical Chemistry and Haematology Values of a Captive Population of Humboldt Penguins (Spheniscus humboldti).

Reference intervals for clinical chemistry and haematology analytes are essential for the interpretation of blood results, but limited data are available for Humboldt penguins (Spheniscus humboldti) in zoological collections as well as in the wild. The aim of the present study was therefore to establish reference intervals for a series of clinical chemistry and haematology analytes in a single zoological population of Humboldt penguins. Furthermore, possible variations of the analytes influenced by the age and sex of the individuals were investigated. Lithium heparinized plasma and whole blood samples from 39 animals were analysed and statistically evaluated. The following clinical chemistry analytes were significantly higher in females: glutamate dehydrogenase (p = 0.0003), alanine transaminase (p = 0.0005), alkaline phosphatase (p = 0.0245), aspartate aminotransferase (p = 0.0043), creatine kinase (p = 0.0016), lactate dehydrogenase (p < 0.0001), inorganic phosphorus (p = 0.0223), and sodium (p = 0.0415). No significant differences between males and females were found for any haematology analyte. Cholesterol (p = 0.0233; -0.39), white blood cell count (p = 0.0208; -0.40), and absolute heterophiles (p = 0.0148; -0.42) had significant negative correlations with the age of the penguins. The results indicate that the factors of sex and age should be considered when interpreting the results of haematological and clinical biochemistry assays, and further studies are needed to determine whether there are more differences in other age groups or seasons.

Comparison of Plasma Trace Elements Between 2 Captive Humboldt Penguin (Spheniscus humboldti) Populations.

Environmental pollutants and their effect on wildlife health play an important role in the conservation of endangered species and can be clinically relevant in captive animals too. Data on relevant concentrations of trace elements in captive birds with no known exposures are rare. For this study, silver, arsenic, gold, barium, cadmium, cobalt, chromium, copper, iron, mercury, magnesium, manganese, molybdenum, thallium, selenium, and zinc were measured by inductively coupled plasma mass spectrometry in lithium heparinized plasma samples from 18 Humboldt penguins at 2 zoological collections in Germany. The results showed that the plasma concentrations of silver, arsenic, gold, cadmium, cobalt, chromium, mercury, and selenium differed significantly (P ≤ 0.05) between the 2 penguin collections. The results indicate that the location of the birds has a strong influence on the plasma trace element concentrations. Well water used in the enclosures was suspected to be associated with these significant differences. Trace elemental concentrations in feed (eg, marine fish) and contamination from enclosure construction materials may also play a role. This study could provide a basis for further comparative, biomonitoring, toxicity, and reference interval studies.

PLASMA VITAMIN AND MINERAL CONCENTRATIONS IN CAPTIVE GREATER FLAMINGOS (PHOENICOPTERUS ROSEUS) AS INFLUENCED BY DIET CHANGE.

The greater flamingo (Phoenicopterus roseus) is often maintained in managed care. Although values for plasma concentrations of minerals and vitamins have been published for this species, limited studies investigate the effects of diet changes on these values. Plasma concentrations of selected vitamins (A [and the provitamin A carotenoid ß-carotene] and E), minerals (Ca, Cu, Fe, K, Mg, Mn, Na, inorganic P, Se, and Zn), and total protein (TP) were measured in a zoo collection of greater flamingos in November 2018 (n = 48) and in November 2019 (n = 52), 30 d after a change from the summer diet, which was the same in both years, to different winter diets. Chloride, cholesterol, and triglycerides were only measured once in 2019. The nutrient profiles of the two complete diets (labeled and analyzed) differed considerably, especially for the canthaxanthin concentration. The plasma concentrations of ß-carotene (P < 0.0001), Cu (P < 0.0001), Fe (P < 0.0001), Mg (P < 0.0001), and Se (P < 0.0001) were significantly higher and vitamin E (P < 0.0001), Ca (P = 0.0014), Mn (P < 0.0001), Na (P = 0.0005), and TP (P < 0.0001) were significantly lower in 2018 than in 2019. Four minerals (Cu, Fe, Mg, and Zn) were measured using both spectrophotometry and inductively coupled plasma mass spectrometry, and all of these minerals showed significant (P < 0.05) differences between the two methods. Based on the results of the study, both the influence of diet and the detection method used for circulating nutrients must be considered when interpreting the plasma vitamin and mineral concentrations in greater flamingos.

COMPARISON OF CLINICAL CHEMISTRY ANALYTES IN GREATER FLAMINGOS (PHOENICOPTERUS ROSEUS) AND AMERICAN FLAMINGOS (PHOENICOPTERUS RUBER) IN A ZOO IN EUROPE.

Reference intervals for clinical chemistry analytes are important for the interpretation of blood results, but reference intervals available in journal articles for specific flamingo collections are often based on a small number of individuals or are only available for a limited number of analytes. Differences in husbandry and nutritional conditions may impact reference intervals. The goal of this study was to establish reference intervals from a mixed population of greater (Phoenicopterus roseus) and American (Phoenicopterus ruber) flamingos when husbandry parameters are equal and to compare the results between the different species, sexes, and age groups in order to evaluate differences between these factors. Lithium-heparinized plasma samples from 93 animals were analyzed for the following: amylase, cholinesterase (CHE), bile acids (BA), total protein (TP), glutamate dehydrogenase (GLDH), aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), uric acid (UA), inorganic phosphorus (P), total calcium (Ca), sodium (Na), and potassium (K). The results show many differences between the species and a few between the different age groups but no differences between males and females of either species. Amylase (P= 0.0010), AST (P= 0.0090), CK (P= 0.0011), and Ca (P= 0.0217) differed significantly between greater and American flamingos. The age group significantly impacted blood levels of CK (P = 0.0299) and LDH (P = 0.0165) in greater flamingos and BA (P = 0.0495), P (P = 0.0422), and Ca (P = 0.0006) in American flamingos. The results show the importance of species-specific reference intervals and the consideration of age-related variations in order to facilitate correct interpretation of blood results.

A pilot study on dietary and faecal calcium/phosphorus ratios in different types of captive ruminating herbivores.

Quantitative differences in calcium and phosphorus metabolism between domestic species exist and can be visualised using data on calcium and phosphorus intake and faecal excretion. The parameter for analysing the results was defined as Δ = dietary calcium/phosphorus (Ca/P) ratio - faecal Ca/P ratio. In previous studies, hindgut fermenters had significantly higher Δ values than ruminants (sheep, cattle, goats), which was explained by the high calcium digestibilities in hindgut fermenters in contrast to highly efficient phosphorus recycling in ruminants. The first hypothesis of the present study was that different types of ruminants (grazer, browser, intermediate feeder) would show differences in Δ as a proxy for quantitative calcium and phosphorus metabolism. The second hypothesis was that camelids as functional, but not taxonomic ruminants would show Δ values similar to ruminants. We used herbivorous zoo animals (17 species, hindgut and foregut fermenters), which were kept on their regular diet without variation for 1 week. Then, faecal samples were obtained from the individual animals. Feed items and faecal samples were analysed for calcium and phosphorus, and dietary and faecal Ca/P ratios as well as Δ were calculated. A comparison of the species groups (one-way ANOVA on ranks, p < 0.05) showed that zoo hindgut fermenters had significantly higher Δ values (0.67 ± 0.34) than camelids and zoo ruminants (-1.07 ± 0.35 and -1.87 ± 1.56). There was no significant difference between camelids, grazers (-1.49 ± 1.31), browsers (-1.63 ± 0.88) and intermediate feeders (-2.11 ± 1.76). The ruminant species from this study had markedly lower Δ than domestic ruminants from literature data. Especially intermediate feeders had low Δ, possibly due to more efficient phosphorus recycling than the domestic ruminants.

Comparison of Capillary Zone Electrophoresis in Greater Flamingos (Phoenicopterus roseus) and American Flamingos (Phoenicopterus ruber).

Electrophoresis can be used to aid in the diagnosis of infectious diseases (eg, aspergillosis) in avian species. Reference intervals for blood plasma proteins of 2 different flamingo species (Phoenicopterus roseus and Phoenicopterus ruber) and their hybrids were calculated by capillary zone electrophoresis (CZE) and differences between these species, sexes, and age groups were evaluated. Lithium-heparinized plasma samples from 111 animals from a zoological collection were analyzed by CZE and statistically evaluated. Differences were only found between greater and American flamingos (P = .003) and between greater flamingos and hybrids (P = .001) in the γ-globulin fraction. Male greater flamingos showed significantly higher α-globulins (P = .022) and females higher total albumin by CZE (P = .037). In American flamingos, the percent total albumin (P = .017), total albumin (P = .025), prealbumin (P = .005), and albumin/ globulin (A/G) ratio (P = .008) were higher in females, and α- (P = .023) and ß-globulins (P = .021) were higher in males of the same species. The following parameters differed significantly between the age groups: γ-globulins (P = .048) in greater and α- (P = .021) and ß-globulins (P = .001) in American flamingos increased with increased age and percent total albumin (P = .002), total albumin (P = .024), and A/G ratio (P = .002) decreased with age in American flamingos. The results showed only small differences between the species, but greater differences between the sexes and ages, especially in American flamingos, which must be considered when interpreting laboratory results.

Topical treatment of multiple erosive, ulcerative skin lesions in an Indian rhinoceros (Rhinoceros unicornis).

A 30-year-old, intact female Indian rhinoceros (Rhinocerus unicornis) was presented with ongoing erosive, ulcerative skin lesions over a 4-year-period. The lesions appeared to be non-pruritic and non-seasonal. A systemic antibiotic therapy had been unsuccessful. The dermatological examination showed 8 diffusely demarcated areas of erosion, focal ulceration, mild crusting, and moderate erythema ranging from 8 to 20 cm in diameter, bilaterally on the lateral edges of the torso armor plates. The patient had no other clinical abnormalities. Coccoid and rod-shaped bacteria were identified on cytology and a bacterial culture revealed Escherichia coli spp., Staphylococcus dysgalacticae, Stenotrophomonas maltophilia, Corynebacterium spp. and Micrococus spp. A topical product line containing essential fatty acids and plant extracts was administered daily, using a spot-on, spray and balm. Within 3 weeks a substantial alleviation of clinical signs was observed. Multiple impression smears of the lesions and a bacterial culture were negative by day 21. The patient achieved complete remission within 4 months of treatment and maintained remission for the 2-year observation period with continued use of the medication. Topically administered therapeutics containing essential fatty acids and plant extracts may offer a viable treatment option for recurrent cases of bacterial infectious skin lesions in Indian rhinocerotidae.

Toxoplasma gondii in small exotic felids from zoos in Europe and the Middle East: serological prevalence and risk factors.

BACKGROUND: Toxoplasma gondii infections and cases of clinical toxoplasmosis have been recorded in zoo animals. Wild felids in human care can serve as definitive hosts that shed oocysts, but also as intermediate hosts for the parasite. Some felid species, such as the Pallas's cat (Otocolobus manul) or sand cat (Felis margarita), may suffer from clinically apparent toxoplasmosis. In the present study, our main aim was to assess risk factors for T. gondii infections in small exotic felids. METHODS: A seroepidemiological study was conducted using the reduviid bug Dipetalogaster maxima for blood sample collection, a method previously evaluated on domestic cats. A total of 336 samples from 17 felid species were collected in 51 institutions, 48 of which were within Europe and the remaining three in the Middle East (United Arabic Emirates and Qatar). These samples were analyzed for T. gondii antibodies by immunoblotting and an immunofluorescent antibody test. Potential risk factors in zoos for seropositivity regarding T. gondii among members of the European Association of Zoos and Aquaria (EAZA) were evaluated using a questionnaire and individual data from the Zoological Information Management System (ZIMS). RESULTS: The sampled felids showed an overall seroprevalence for T. gondii of 63%. The risk factor study including data of 311 small exotic cats of 10 species resulted in a final generalized linear mixed model comprised of five variables: the likelihood of seropositivity increased statistically significantly with "Age", while feeding "Cattle: frozen" relative to "Cattle: fresh", "Outdoor housing fenced in on all sides", "Mesh size 2-5 cm" relative to "Mesh size > 5 cm" and "Wearing gloves: yes" had statistically significant protective effects. CONCLUSIONS: Wild felids, including endangered species, kept in human care in European and Middle Eastern institutions, are widely exposed to T. gondii. Risk factor analysis revealed that feeding previously frozen tissues, keeping animals in enclosures that are fenced on all sides using fences with small mesh sizes, and wearing gloves when working inside enclosures seem to be the most relevant protective measures to prevent T. gondii infections in these animals .