Hematological and serum biochemical parameters of rescued Sunda pangolins (Manis javanica) in Singapore.

The Sunda pangolin (Manis javanica) population in Southeast Asia faces threats such as poaching and deforestation. Health assessments of rescued individuals including physical examination and blood work are crucial for clinicians to determine the health status of these animals. The establishments of reference intervals of hematology and serum biochemistry are important for identifying clinical abnormalities. The objectives of our study were to establish blood reference intervals for Sunda pangolins, to determine if there are age and sex related differences in hematology and serum biochemistry, and to compare our results with those of a previous study on confiscated Sunda pangolins in Thailand. Fifty-eight Sunda pangolins were rescued between January 2011 and December 2015. The hematology and serum biochemistry results of 51 clinically normal Sunda pangolins were selected for the establishment of the blood reference intervals. No sex related differences were noted in this study. Age-related differences were observed, in which adult Sunda pangolins had a significantly higher mean corpuscular volume than juveniles, and juvenile Sunda pangolins had significantly higher red blood cell counts and hemoglobin levels than those of the adults (P<0.05). Age-related differences were also noted in several serum biochemistry parameters: alkaline phosphatase (ALP) was significantly higher in juveniles, and total protein was significantly higher in adult Sunda pangolins. Compared to a previous study the white blood cell counts, neutrophil counts, and ALP were higher, and the lymphocyte counts were lower in the present study.

Biochemical parameters of the giant anteater (Myrmecophaga tridactyla Linnaeus, 1758) of the Brazilian Cerrado / Parâmetros bioquímicos de tamanduás-bandeiras (Myrmecophaga tridactyla Linnaeus, 1758) do cerrado brasileiro

The giant anteater (Myrmecophaga tridactyla) is classified as a vulnerable species on Brazil's list of species at risk of extinction mainly due to deforestation and forest fires. This has contributed to a considerable increase in detailed clinical case records of the treatment of wild species at veterinary institutions. However, the paucity of serum biochemical profiles of healthy giant anteaters has made it difficult to evaluate these animals, preventing diagnosis, treatment and prognosis. The objective of this work was to collect data about the biochemical profile of healthy giant anteaters from the Brazilian Cerrado raised in captivity, in order to better understand the physiological characteristics inherent to this species. Eighteen analytes from 12 healthy giant anteaters were measured. The following means and standard deviations were found in the biochemical analyses: albumin 3.29±0.33g/dL, ALT 15.49±7.98 IU/L, amylase 1037.92±149.04 IU/L, AST 21, 12±7.50 IU/L, total cholesterol 62.79±20.08mg/dL, HDL cholesterol 14.73±4.98mg/dL, LDL cholesterol 26.60±11.05mg/dL, VLDL cholesterol 2.14±1.06mg/dL, CK 111.61±70.16 IU/L, creatinine 1.05±0.37mg/dL, iron 194.64±81.17µg/dL, GGT 65.18±54.57 IU/L, glucose 103.71±29.63mg/dL, globulins 2.76±0.36g/dL, lipase 28.80±5.11 IU/L,TSP 6.05±0.56g/dL, triglycerides 10.71±5.29mg/dL, and urea 53.46±18.28mg/dL. The values found in this study can be used as references for the laboratory evaluation of giant anteaters living in conditions similar to those of this study. This is one of the first reports of biochemical examinations on giant anteaters of the Cerrado biome.(AU)

O tamanduá-bandeira (Myrmecophaga tridactyla) está classificado como espécie vulnerável na lista brasileira de espécies ameaçadas de extinção devido principalmente ao desmatamento e aos incêndios florestais. Tal fato contribuiu com o aumento da casuística de atendimento de espécies silvestres em instituições veterinárias. Porém, a escassez de valores bioquímicos séricos em tamanduás-bandeiras hígidos tem dificultado a avaliação destes animais, impedindo o diagnóstico, tratamento e prognóstico. O objetivo deste trabalho foi fornecer dados sobre o perfil bioquímico de tamanduás-bandeiras saudáveis do cerrado brasileiro, criados em cativeiro, a fim de compreender melhor as características fisiológicas inerentes a esta espécie. Foram mensurados 18 analitos de 12 tamanduás-bandeiras hígidos. As médias e o desvio padrão correspondentes às análises bioquímicas foram: albumina 3,29±0,33g/dL; ALT 15,49±7,98 UI/L; amilase 1037,92±149,04 UI/L; AST 21, 12±7,50 UI/L; colesterol total 62,79±20,08mg/dL; colesterol HDL 14,73±4,98mg/dL; colesterol LDL 26,60±11,05mg/dL; colesterol VLDL 2,14±1,06mg/dL; CK 111,61±70,16 UI/L; creatinina 1,05±0,37mg/dL; ferro 194,64±81,17µg/dL; GGT 65,18±54,57 UI/L; glicose 103,71±29,63mg/dL; globulinas 2,76±0,36g/dL; lipase 28,80±5,11 UI/L; PST 6,05±0,56g/dL; triglicerídeos 10,71±5,29mg/dL; ureia 53,46±18,28mg/dL. Os valores encontrados neste estudo podem ser utilizados como referência para a avaliação laboratorial de tamanduás-bandeiras que vivam em condições similares ao do presente estudo. Este é um dos primeiros estudos a relatar exames bioquímicos em tamanduás-bandeiras do bioma cerrado.(AU)

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)

A successful transfusion in a tamandua (Tamandua tetradactyla) using both whole blood and blood replacement products.

A 10-yr-old tamandua (Tamandua tetradactyla) presented with severe lethargy and ataxia. Severe anemia (9% hematocrit) was diagnosed and ultimately resolved with the administration of a whole blood transfusion from conspecific and additional blood replacement products (Oxyglobin). This is the first described use of a whole blood transfusion in an edentate.

Hematology values of captive giant anteaters (Myrmecophaga tridactyla) and collared anteaters (Tamandua tetradactyla) / Valores hematológicos de tamanduá-bandeira (Myrmecophaga tridactyla) e tamanduá-mirim (Tamandua tetradactyla) oriundos de cativeiro

Hematological results are reported for 13 giant anteaters (Myrmecophaga tridactyla) and 13 collared anteaters (Tamandua tetradactyla). Animals were captive-reared adults held at the Fundação Parque Zoológico de São Paulo (São Paulo, SP, Brazil) and Parque Zoológico Municipal Quinzinho de Barros (Sorocaba, SP, Brazil), and were considered healthy on physical examination. Examined parameters included red blood cell count, white blood cell count, hematocrit, hemoglobin concentration, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, total plasmatic protein and differential leukocyte counts. Also, a survey for hemoparasites was done and none was observed in thin blood smears. The results were generally similar to those previously reported in the exiguous literature for these species, providing further reference data for the interpretation of laboratory results besides health monitoring, assisting early disease diagnosis and providing relevant information for conservation programs for these species.

Obteve-se os valores hematológicos de 13 tamanduás-bandeira (Myrmecophaga tridactyla) e 13 tamanduás-mirim (Tamandua tetradactyla) adultos e saudáveis segundo exame físico, oriundos de cativeiro, da Fundação Parque Zoológico de São Paulo (São Paulo) e do Parque Zoológico Municipal Quinzinho de Barros (Sorocaba). Os parâmetros hematológicos estudados incluíram contagem total de eritrócitos e leucócitos, hematócrito, concentração de hemoglobina, volume corpuscular médio, hemoglobina corpuscular média, concentração de hemoglobina corpuscular média, proteína plasmática total estimada e contagem diferencial de leucócitos. Também se realizou a pesquisa de hemoparasitas, não se encontrando nenhum nos esfregaços sanguíneos. De maneira geral, os valores obtidos não diferiram muito daqueles presentes na pouca literatura existente, contribuindo com dados adicionais para a interpretação de resultados laboratoriais, visando acompanhamento do estado sanitário, auxílio no diagnóstico de enfermidades e programas de conservação dessas espécies.

Leptospira and Brucella antibodies in collared anteaters (Tamandua tetradactyla) in Brazilian zoos.

The presence of Leptospira spp. and Brucella spp. antibodies was investigated in serum samples from 28 collared anteaters (Tamandua tetradactyla) kept in seven Brazilian zoos. Sera were tested against 19 Leptospira serovars using microscopic agglutination. Samples reacted to the following serovars: two (7.14%) to Patoc, three (10.71%) to Tarrasovi, three (10.71%) to both Patoc and Tarrasovi, two (7.14%) to Wolffi, and one (3.57%) to Australis. Two (7.14%) samples reacted to the buffered Brucella antigen test, but no confirmatory reaction occurred using the 2-mercaptoethanol slow slide agglutination test. No sample was reactive in the agar gel immunodiffusion test for rugose species of Brucella. The presence of anti-leptospira agglutinins in captive T. tetradactyla serum indicates that this species may be susceptible to infection by these bacteria.

Relationship between gonadal steroid hormones and vulvar bleeding in southern tamandua, Tamandua tetradactyla.

This study aimed at demonstrating the profiles of circulating gonadal steroid hormones during the estrous cycle and pregnancy in a southern tamandua (Tamandua tetradactyla). Additionally, this study clarified the relationship between vulvar bleeding and hormonal changes. The concentrations of serum progesterone (P(4)) and estradiol-17ß (E(2)) were determined by enzyme immunoassays. Serum P(4) and E(2) concentrations changed cyclically and the estrous cycle length (± SD) based on the E(2) cycles was 44.3 ± 4.5 days. Vulvar bleeding started to be seen at the decreasing of P(4). The cycle length for vulvar bleeding was 43.3 ± 4.2 days. Interval from the first day of bleeding to the peak of E(2) concentration was 23.1 ± 3.1 days. Serum P(4) during pregnancy remained high and E(2) increased 8 weeks after conception and remained high until parturition. The female delivered normally after a 165 day-pregnancy period and reared the offspring well. Approximately 3 weeks after parturition, serum E(2) and P(4) cycles resumed. Visual bleeding may be useful as a real-time indicator for understanding the ovarian cycle of southern tamanduas, and estrus could be expected approximately 3 weeks after the first bleeding.

Evidence on mammalian phylogeny from sequences of exon 28 of the von Willebrand factor gene.

Phylogenetic relationships among 27 extant mammalian species (representing 15 placental orders) were studied using sequences of exon 28 of the gene encoding von Willebrand Factor (vWF), a glycoprotein which functions in blood clotting. Analysis of sequences coding for vWF revealed evidence for several subordinal and superordinal groupings, but the earliest branching sequence of placental mammals was left largely unresolved. Strong support was found for a monophyletic clade consisting of elephants, sea cows, hyraxes, aardvarks, and elephant shrews. This systematic placement of the elephant shrews agrees strongly with two other molecular data sets (interphotoreceptor retinoid binding protein and alpha-lens crystallins) and is consistent with analysis of fossil elephant shrews recently discovered in north Africa. Evidence from vWF sequences agrees with a number of previous molecular and morphological studies in providing strong support for the monophyly of both bats and rodents. The orders Primates, Proboscidea, Carnivora, Perissodactyla, and Artiodactyla were represented by more than one species which joined in each case to form a monophyletic order.

Plasma and erythrocyte cholinesterase values for the common long-nosed armadillo, Dasypus novemcinctus.

Plasma and erythrocyte cholinesterase activities were determined for 40 free-living and 12 captive common long-nosed armadillos (Dasypus novemcinctus) in order to establish normal values for monitoring pesticide exposure. Plasma cholinesterase activity ranged from 105 to 549 U/liter with no sexual or seasonal differences. Plasma values from captive animals were significantly lower than those from wild armadillos. Erythrocyte cholinesterase activity ranged from 2,915 to 15,126 U/liter with no differences detected between captive and wild animals or between sexes. However, erythrocyte cholinesterase values varied seasonally. Erythrocyte and plasma cholinesterase activities were not significantly correlated. Packed cell volume ranged from 24 to 51% and did not vary significantly between captive and wild samples, between sexes or among seasons. However, both whole blood and erythrocyte cholinesterase activities showed significant negative correlations with packed cell volume. Controlled experiments are needed to find the factors responsible for the statistically significant difference between plasma cholinesterase activities of captive and wild armadillos. The seasonal variation in erythrocyte cholinesterase activity and the negative correlation between erythrocyte cholinesterase activity and packed cell volume can be explained by an hypothesis that relates the variation in erythrocyte cholinesterase activity to variation in erythrocyte turnover rate. Future work should involve experiments to test this hypothesis.

The primary structure of pale-throated three-toed sloth (Bradypus tridactylus, Xenarthra) hemoglobin.

The hemoglobin of the Pale-Throated Three-Toed Sloth (Bradypus tridactylus, Xenarthra) was separated into two components (ratio 4:1) with identical amino-acid analyses for the alpha- and beta-chains. The primary structures of both chains from the major component are given. They could be isolated by chromatography on carboxymethyl cellulose CM-52. The sequences have been determined by automatic Edman degradation of the native chains and their tryptic peptides. The comparison with human hemoglobin showed 27 substitutions in the alpha-chains and 33 in the beta-chains. In the alpha-chains one amino-acid exchange involves an alpha 1/beta 1-contact. In the beta-chains two heme- and four alpha 1/beta 1-contacts are substituted. The hemoglobin of the Sloth is compared to that of the Nine-Banded Armadillo (Dasypus novemcinctus), another representative of the order Xenerthra.

Oxygen binding functions of blood and hemoglobin from the Chinese pangolin, Manis pentadactyla: possible implications of burrowing and low body temperature.

We measured O2 equilibria of adult blood and of 'stripped' (cofactor-free) hemolysates from adult and newborn Manis pentadactyla, in order to assess the implications of the burrowing habit and the low deep-core temperature in pangolins, and to discern the mechanisms for maternal-fetal O2 transfer. At pH 7.4 and body temperature (33 degrees C) the blood O2 affinity was significantly higher than in similarly sized non-burrowing, 'normothermic' mammals (P50 = 21 and 33 mm Hg, respectively) indicating an adaptation to hypoxic burrow conditions. This difference is not attributable to a higher intrinsic O2 affinity in the pangolin Hb or to significant differences in its sensitivity to temperature and erythrocytic 2,3 diphosphoglycerate (DPG), but tallies with lower DPG/Hb ratios than generally found in mammals. Stripped adult and newborn hemolysates show similar O2 affinities and pH and DPG sensitivities, but reveal a specific adult Hb that develops after birth, in sharp contrast with the ontogenetic changes in other mammals where specific fetal Hbs are lost after birth.

Fetal-maternal gradients of steroid hormones in the nine-banded armadillo (Dasypus novemcinctus).

Gonadal and adrenal steroid hormone levels in plasma were measured in pregnant, non-pregnant and fetal armadillos. Progesterone demonstrated the largest placental gradient with levels 6 to 10-fold higher in the fetus. Testosterone levels were higher in male and lower in female fetuses, when compared to the respective maternal serum values. Dehydroepiandrosterone, and cortisol were consistently high in maternal as compared to matched fetal serum samples whereas the reverse was true for androstenedione. Tissue incubations with labelled pregnenolone indicated the major source of progesterone production to be the fetal adrenal whereas the fetal testes demonstrated a major conversion product consistent with dehydroepiandrosterone.

Receptor in group C and G streptococci detects albumin structures present in mammalian species.

The presence of albumin structures with the capacity to bind to a surface receptor in group C and G streptococci was studied in serum samples from 45 mammalian species representing 15 different orders, using an inhibition assay. The ability of animal sera to inhibit the uptake of radiolabeled human serum albumin by the streptococci indicated the presence of such albumin structures. Positive reactions were found in species of most orders tested, with Marsupialia as a notable exception. All Carnivora sera tested were strongly positive. In some orders such as Artiodactyla both positive and negative species were identified. Serum samples from 62 bird species representing 15 orders and from 5 fish species were also tested in the inhibition assay. None of these serum samples was capable of inhibiting the uptake of human serum albumin by streptococci. Some differences were also noted in the results obtained with group C and G streptococci from human and bovine sources, respectively, indicating the presence of two types of receptors. The present studies suggest a phylogenetic origin of albumin structures with affinity for the streptococcal receptor to a period after the divergence of Marsupialia from the other mammalian orders.

[Hemoglobins XLVI: the primary structure of the alpha-chain of armadillo (Dasypus novemcinctus, Edentata) hemoglobin (author's transl)]. / Hämoglobine, XLVI. Die Primärstruktur der alpha-Ketten des Hämoglobins vom Gürteltier (Dasypus novemcinctus, Edentata).

The complete primary structure of the identical alpha-chains of the two hemoglobin components of armadillo (Dasypus novemcinctus) is presented. It was established on the tryptic peptides by automatic Edman degradation. The alignment was done according to the homology with human alpha-chains. 25 differences were found between both chains. A comparison of the functional amino acid residues shows one substitution in the surrounding of the heme, there in the alpha 1 beta 1 - and two in the alpha 1 beta 2 - subunit interface. The two replacements alpha 38(C3)Thr leads to Pro and alpha 44(CD) - Pro leads to Ser may contribute to the high oxygen affinity of the armadillo hemoglobin by destabilization of the T-structure.

Annual circulating testosterone levels in captive and free-ranging male armadillos (Dasypus novemcincuts).

Mean circulating concentrations and seasonal changes in plasma testosterone over 3 years were similar in free-ranging and captive male armadillos. The highest concentrations were seen in the summer months, i.e. at the time of maximum breeding, but the relatiely high values at other times and the inability of ACTH to restore dexamethasone-suppressed testosterone concentrations confirms that testicular function continues throughout the year. There was a gradual rise in plasma testosterone to adult levels from the 5th to the 9th month of life in captive-born males, indicating that sexual maturity is reached within the first year of life and that captivity is not deleterious to Leydig cell function.

Hematologic parameters for wild and captive nine-banded armadillos (Dasypus novemcinctus).

Hematologic parameters were determined at capture and during captivity for nine-banded armadillos. Compared to the time of capture, the mean erythrocyte counts were significantly lower after 6 months in captivity. After 1 year in captivity, the mean hematocrit was significantly higher and the mean neutrophil count in males was significantly lower. Although the number of armadillos sampled was greater than in previous research, the variation within groups was large.