Serum biochemistry of the Arrau turtle (Podocnemis expansa): Frozen analyte stability and the effects of long-term storage.

BACKGROUND: Turtles are a major source of protein for riverside human populations in Brazil. The encouragement of commercial breeding meets conservation efforts for these animals, and it is, therefore, crucial to understand the physiologic and behavioral aspects of semi-aquatic species in captive conditions. Serum biochemical tests are ancillary diagnostic tools, and sample storage is a main problem since clinical laboratories are not always available near the habitats of these species. OBJECTIVES: The aim of this study was to provide information about the stability of albumin, aspartate aminotransferase (AST), calcium, creatinine kinase (CK), total cholesterol (Chol), alkaline phosphatase (ALP), gamma glutamyltransferase (GGT), total protein (TP), and urea at different storage times. METHODS: In all, 17 Arrau turtles (Podocnemis expansa) were used, and the serum obtained was separated into aliquots and analyzed at 0, 4, 8, 16, and 32 days after being stored at -20°C. RESULTS: The results showed that albumin, AST, CK, GGT, and TP suffered interference due to the long storage times. CONCLUSION: Analytes such as ALP, calcium, Chol, and urea can be evaluated for up to 1 month after freezing. Albumin, AST, and TP can be analyzed up to 1 week after freezing without alterations, and CK GGT are best evaluated on fresh samples.

Giant Anteater (Myrmecophaga tridactyla Linnaeus, 1758) of the Brazilian cerrado: hematology and storage effect / Tamanduás-bandeiras (Myrmecophaga tridactyla Linnaeus, 1758) do cerrado brasileiro: hematologia e efeito de estocagem

Giant Anteater (Myrmecophaga tridactyla) is a vulnerable species because of progressive habitat destruction, mostly affected by wildfires and car accidents. The increasing number of animals that are attended by wildlife rescue centres reinforces the need of knowledge about haematological parameters, especially for those that inhabit Brazilian cerrado biome. For this purpose and in order to establish reference values for this species and also to compare them with previous studies, haematological analysis of captive giant anteaters from Brazilian cerrado were performed. Moreover, the alterations of blood samples after 24 and 48 hours of storage at refrigeration temperatures (4oC) and preserved with two different EDTA concentrations (5% and 10%) were studied. Means and standard deviations of haematological parameters analysed immediately after collection were: RBC: 2,07 x106/µL ± 0,40; hematocrit: 38,08%± 5,93; haemoglobin: 11.33g/dL±2.15; MCV:186.52 fL±21.72; MCHC: 29.68g/dL±2.56; MCH: 55.08pcg±5,94; total leucocytes: 8.142/µL±2.441; neutrophils: 5.913/µL±2.168; lymphocytes: 1.460/µL±740; eosinophil: 522/µL±385; monocytes: 247/µL±176; thrombocytes: 123.458/µL±31.362 and total plasma protein: 6.23g/dL±0.49. This data shows evidence of the existence of important differences between these values and others from other areas, either from Brazil or from other South American countries. Those variations might be connected to environment, genetic, nutritional and/or management factors. Regarding the storage effect analysis, it can be concluded that in giant anteaters, haematological analysis can be performed until 24h after collection without any significant alterations on the haematological parameters, except for thrombocytes. Concerning the different EDTA concentrations, it can be concluded that there are no quantitative differences in haematological variables. Nevertheless, relevant morphologic alterations in blood cells can be observed after a 24h storage period, being most noticeable in the leucocytes. Those alterations can lead to misinterpretation of the results, interfering diagnosis, prognosis and treatment.(AU)

O tamanduá-bandeira (Myrmecophaga tridactyla) é uma espécie vulnerável devido à destruição progressiva do seu habitat natural, sendo afetado por queimadas e atropelamentos. O aumento na casuística de atendimentos de animais silvestres reforça a necessidade de se ter conhecimento dos parâmetros hematológicos, em especial para os que vivem no bioma do cerrado. Por isso, este trabalho teve por objetivos realizar o hemograma de tamanduás-bandeiras de cativeiro localizados no cerrado brasileiro, a fim de estabelecer valores de referência para essa espécie e compará-los a estudos prévios. Além disso, verificar quais alterações podem ser encontradas em amostras armazenadas por 24 e 48 horas após a colheita, em temperatura de refrigeração (4oC) e tratadas com duas concentrações distintas de EDTA (5% e 10%). A média e o desvio padrão das variáveis hematológicas encontradas nas amostras processadas logo após a colheita foram: hemácias (2,07x106/µL±0,40); volume globular (38,08%± 5,93); hemoglobina (11,33g/dL±2,15); VCM (186,52 fL±21,72); CHCM (29,68g/dL±2,56); HCM (55,08pcg±5,94); leucócitos totais (8.142/µL±2.441); neutrófilos (5.913/µL±2.168); linfócitos (1.460/µL±740); eosinófilos (522/µL±385); monócitos (247/µL±176); plaquetas (123.458/µL±31.362) e proteínas plasmáticas totais (6,23g/dL±0,49). Tais dados permitem afirmar que existem importantes diferenças entre os valores hematológicos destes em relação aos animais provenientes de outras regiões, tanto do Brasil quanto de outros países da América do Sul. Provavelmente, tais divergências estão associadas a fatores ambientais, genéticos, nutricionais e/ou de manejo. Quanto à análise das amostras estocadas, conclui-se que, em tamanduás-bandeiras, as amostras para a realização de hemograma podem ser processadas até 24 horas após a colheita, sem alteração significativa das variáveis hematológicas, com exceção das plaquetas. Com relação às duas concentrações de EDTA, pode-se inferir que não há diferença quantitativa entre ambas para as variáveis hematológicas. Contudo, é possível constatar que o EDTA promove alterações morfológicas relevantes nas células sanguíneas após 24 horas de armazenamento, sendo os leucócitos os mais afetados. Tais alterações, quando relatadas, podem induzir interpretações equivocadas, interferindo no diagnóstico, prognóstico e tratamento.(AU)