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.

Cutaneous apocrine gland neoplasia in 16 captive African wild dogs (Lycaon pictus).

This retrospective study documents the occurrence of single and multiple cutaneous apocrine gland tumours (CATs) on the dorsal midline of 16 captive African wild dogs (AWDs, Lycaon pictus) derived from 161 submissions to diagnostic laboratories in South Africa, France and Germany between 1997 and 2022. Animals included in the study came from zoological institutions in South Africa (n = 2), France (n = 5) and Germany (n = 1) and ranged from 5 to 14 years of age. Fifteen affected animals were female (94%) and one was male. CATs presented as raised, hairless, multilobular, grey firm masses, consistently located along the dorsal midline. Apart from a single cutaneous apocrine adenoma and a cystadenoma occurring concurrently with two non-cystic adenocarcinomas, neoplasms were consistent with malignant cutaneous apocrine adenocarcinomas with lymphatic spread and visceral metastases. Advanced age and female sex were identified as risk factors. A genetic component or association with the increasing use of GnRH agonist contraceptives was suspected but could not be established. This study highlights the need for close clinical monitoring of AWDs over the age of 5 years for the development of CATs along the dorsal midline and supports early surgical intervention. More research is needed to determine the role of inbreeding, endocrine changes and husbandry factors that may play a role in the development of CATs on the dorsal midline of AWDs.

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.

Demodex carolliae in a colony of Seba's short-tailed bats (Carollia perspicillata): clinical, pathological and parasitological findings.

Seba's short-tailed bats (Carollia perspicillata) are a frugivorous species native to Central and South America. Despite their importance as a reservoir for zoonotic pathogens and their popularity in zoological collection and as research models, there are relatively few reports on non-zoonotic diseases of bats. Mites of the genus Demodex are obligate commensals of the skin of a range of mammals, are highly host-specific and are not associated with clinical disease when present in low numbers. However, infestation with high numbers can result in severe or even fatal disease and substantially affect the well-being of the animals. The clinical, pathological and parasitological findings in 12 Seba's short-tailed bats with demodicosis from a colony kept at Munich Zoo Hellabrunn between 1992 and 2021 are described in this report. From 2002, skin lesions became apparent on the head, especially the periocular region, nose and ears, as well as the genital area of some animals. In advanced cases, skin changes were also present on the abdomen, back and extremities. Gross findings typically included alopecia and thickening of the skin, with the formation of papules, reflecting cystically dilated hair follicles containing myriads of demodecid mites. Histologically, lesions were characterized by a paucicellular lymphocytic dermatitis and folliculitis with perifollicular fibrosis, epidermal hyperplasia, orthokeratotic hyperkeratosis and disproportionately high numbers of intrafollicular arthropods. Demodex carolliae was identified morphologically by light, phase-contrast and electron microscopy. Further characterization was achieved by extraction of parasitic DNA and partial gene sequencing of two mitochondrial genes, 16S rDNA and cox1. This is the first clinicopathological description of generalized demodicosis in Seba's short-tailed bats and includes the first molecular characterization of D. carolliae with provision of a GenBank entry.

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.

Endodontic Treatment of a Traumatic Tusk Fracture With Exposed Pulp in an Asian Elephant (Elephas maximus).

Tusk fracture in elephants is a common incident often resulting in pulp exposure and pulpitis. Extensive lavage, endodontic therapy, direct pulp capping, or extraction are treatment options. In this report, the successful management of a broken tusk of a juvenile male Asian elephant (Elephas maximus) including morphological analysis of the tusk tip 2 years after surgery are presented. Treatment was carried out under barn conditions and included antimicrobial photodynamic therapy and partial pulpotomy with direct pulp capping. Immediate pain relief was reached. The fractured tusk was preserved and continued to grow. The therapeutic filling material remained intact for over 1 year but was absent 2 years after treatment. The former pulp cavity of the tusk tip was filled with reparative dentin, osteodentin, and bone, but the seal between these hard tissues and pulp chamber dentin was incomplete. Radiographs obtained 3 years after treatment showed no differences in pulp shape, pulp width, and secondary dentin formation between the treated right and the healthy left tusk. It can be concluded that in case of an emergency, the endodontic therapy of a broken elephant tusk can be attempted under improvised conditions with adequate success. Photodynamic therapy might contribute to prevent infection and inflammation of the pulp. The decision tree published by Steenkamp (2019) provides a valuable tool to make quick decisions regarding a suitable therapy of broken tusks.

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.

Glucocorticoid treatment of ovariectomized sheep affects mineral density, structure, and mechanical properties of cancellous bone.

UNLABELLED: Thus far, orthopedic research lacks a suitable animal model of osteoporosis. In OVX sheep, 6 months of steroid exposure reduced bone density and mechanical competence. Bone properties and bone formation did not recover for another 6 months. Therefore, steroid-treated OVX sheep may serve as a large animal model for osteopenic bone. INTRODUCTION: The purpose of this study was to explore the effects of glucocorticoid treatment on cancellous bone density, microarchitecture, biomechanics, and formation of new bone. MATERIALS AND METHODS: Sixteen ovariectomized merino sheep received either a 6-month glucocorticoid treatment (GLU; 0.45 mg/kg methylprednisolone) or were left untreated (control). Cancellous bone biopsy specimens from the tibia were harvested 6 months after ovariectomy. After 12 months, the animals were killed, and biopsy specimens were obtained from the contralateral tibia and the lumbar spine. All biopsy specimens were scanned for apparent bone mineral density by peripheral quantitative computed tomography (pQCT) and tested mechanically in uniaxial compression. Three-dimensional bone reconstructions were obtained by microcomputed tomography. Formation of new bone was analyzed using histologies of the femoral condyles. RESULTS: After 6 months, mineral density (-19%) and mechanical competence (-45%) were reduced by glucocorticoid treatment (p < 0.1). BV/TV (-21%; p < 0.01) and trabecular thickness (-20%; p = 0.01) declined, whereas BS/BV increased (24%; p = 0.01). After 12 months, mineral density (-33%) and mechanical properties (-55%) were reduced even more profoundly (p < 0.05). Also, the structural parameters (BS/BV and Tb.Th.) still seemed to be affected by glucocorticoid treatment (p < 0.05). New bone formation, assessed by measurement of osteoid surface, was markedly reduced (-63%, p < 0.1) by glucocorticoid treatment. The differences between groups were generally more pronounced at the tibia and the femur than at the spine. CONCLUSION: The effects of short-term high-dose steroid administration on bone mineral in this animal model were comparable with those observed in humans after long-term corticoid treatment. Reduction in bone quality and bone formation rate persisted after the cessation of steroid administration. Glucocorticoid treatment of ovariectomized sheep may therefore serve as a large animal model for steroid-induced osteopenia.

Glucocorticoid-treated sheep as a model for osteopenic trabecular bone in biomaterials research.

The purpose of this study was to determine the alterations in ovine trabecular bone induced by a combination of ovariectomy and steroid treatment. Twenty-four female skeletally mature Merino sheep were randomly assigned to ovariectomy alone (OVX), ovariectomy combined with glucocorticoid treatment for 6 months (OVX + GC), or no treatment (control). Biopsies of trabecular bone were harvested 6 and 12 months after the beginning of the study from the proximal tibia. The biopsies were scanned for apparent bone mineral density by quantitative computed tomography and were mechanically tested. Three-dimensional bone reconstructions were obtained by micro-computed tomography. Trabecular bone from the OVX + GC animals had a markedly reduced apparent bone mineral density (27% less than control), bone volume (34%), and elastic modulus (36%) at 6 months. At 12 months, the reductions in apparent bone mineral density (33%), bone volume (37%), and elastic modulus (62%) appeared to be even more pronounced. Ovariectomy alone did not result in a perceptible reduction in any parameter. The combination of ovariectomy and glucocorticoid treatment in sheep resulted in a successful induction of substantial loss of trabecular bone and thus may serve as a large-animal model for osteopenic trabecular bone for the development and testing of orthopedic implants and techniques under osteoporotic conditions.