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Improving the re-introduction of rescued Magellanic penguins through extended health evaluation with minimal sampling volume and material

Julia B. Morais1*, Julieta Volpato1, Adson Costa1, Cristiane Kolesnikovas2, Matheus De Souza3 and Mere E. Saito1
  • 1 UDESC - Universidade do Estado de Santa Catarina, Veterinary Medicine, Brazil
  • 2 R3 Animal - NGO, Veterinary Medicine, Brazil
  • 3 Centro de Triagem de Animais Silvestres, CETAS, Brazil

Debilitated Magellanic penguins, Spheniscus magellanicus, are frequently rescued in the coast of Santa Catarina and taken to rehabilitation centers, where they are clinically treated and evaluated for re-introduction. This species is considered by the IUCN red list as Near Threatened, therefore, its adequate reintroduction in the wild is an important conservation step (Borboroglu & Boersma, 2015). Although the program of rehabilitating and banding penguins was already applied to more than 1100 exemplars, less than 100 have been re-sighted, posing a challenge to the current re-habilitation and re-introduction practitioners (Niemeyer et al, 2010). Currently, the health parameters for penguin re-introduction, adopted by the CETAS (Screening Center of Wild Animals) of Florianópolis, are based on the criteria of the International Found for Animal Welfare (IFAW). This includes good general condition of the animals, waterproofing of the feathers, minimum hematocrit of 38%, maximum buffy coat 2% of the capillary tube and total plasma protein concentration of more than 3 g/dL. However, these parameters do not fully reflect the health status of the animals and are from other rehabilitation institutions or based on studies in other species. Biochemical analysis is an important tool to assist in the health evaluation and disease diagnostics of wild birds because the clinical signs in these animals are often inexpressive (Coles, 1986; Hochleithner, 1994; Schmidt et al., 2007). While Coraiola (2012) refers several biochemical parameters useful in evaluating the rehabilitation of Magellanic penguins, to the author’s knowledge, this practice is still not routinely executed. A field manual compiled by Brazilian environmental and marine authorities proposed standardizing the diagnostic methods applied in penguin rehabilitation facilities. These guidelines recommend hematology be performed in plasma with heparin or EDTA and biochemistry analysis in serum (Vanstreels et al, 2012). As the rescue penguin is usually juvenile, a small animal with less than 4.5 kg, the collection of large amounts of blood may delay its recovery and/or impair its successful return to the colony. A serum sample always yields lower volumes than the plasma one. Since anticoagulants are necessary for hematologic evaluation, it was considered useful to test the viability of plasma for biochemistry analysis of these animals. In this work, we analysed the serum and plasma with two anticoagulants, EDTA 3% and sodium heparine, of 34 rescued penguins from the CETAS of Florianópolis, to compare the blood biochemistry values of 16 analytes (Figure 1). The following analytes were assayed: uric acid, urea, total protein, albumin, globulins, glucose, cholesterol, triglycerides, gamma-glutamyl transpeptidase (GGT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), alkaline phosphatase (FA), lactate dehydrogenase (LDH) and free hemoglobin (Hb). Hemolysis is known to affect biochemistry spectrophotometric readings and alter blood constituents, so, we measured the concentration of free hemoglobin in all samples and excluded the results outside of referenced Hb limits proposed by either the reagent manufacturer or previous avian biochemistry studies (Hawking, 2006). Biochemistry values from animals with visible lesions were analyzed separately. Results showed that, although not statistically significant, samples anticoagulated with EDTA had a higher median of free hemoglobin concentration than serum and heparinized plasma. Therefore, EDTA seemed less efficient than heparin in minimizing in vitro hemolysis. In addition, we reviewed the literature and noticed that heparinized plasma is increasingly being used for biochemistry analysis of penguins (Parsons et al, 2015). Also, the reference intervals provided by the Association of Zoos and Aquariums, AZA, were produced using a large number of same-species animals regardless of sample type (Teare, 2013). Regarding the mean/median values of biochemistry analytes measured in heparinized plasma and plasma with EDTA compared to serum, a smaller number of statistical differences were found between heparinized samples and serum (albumin, globulins, triglycerides and cholesterol) than between samples with EDTA and serum (total protein, globulins, alkaline phosphatase, cholesterol, glucose, uric acid, GGT, ALT and CK). Moreover, when comparing our results of healthy individuals with the reference values proposed by AZA, it was possible to verify that the mean values of globulins (in serum) and alkaline phosphatase (in plasma containing EDTA) were outside the established reference intervals. On the other hand, all mean/median values measured in heparinized plasma fell within the AZA reference intervals. Nevertheless, it was observed that the mean globulin concentration obtained in this study in all sample types was relatively high comparing to the reviewed papers with published globulin concentration levels in S. magellanicus, even after excluding the animals with visible lesions (Campos et al, 2014; Coraiola, 2012; Ghebremeskel et al, 1989). It is known that during rehabilitation many penguins acquire diseases atypical in the wild, like avian malaria, aspergillosis, pox virus infections and bumblefoot. While it is possible that our results were associated with the presence of sub-clinical inflammations, it was not possible to conclude the cause of such globulin increase. According to Villouta et al (1997) studies in Humboldt penguins, S. humboldti, a period of more than 3 weeks of confinement lead to an increase in total proteins and globulins concentrations. The author discarded inflammation as a possible cause for this increase because fibrinogen measurements remained stable with increased captivity time. We suggest the inclusion of fibrinogen measurement, which can only be done in plasma samples, along with the studied analytes, for a more sucessful re-introduction of the Magellanic penguin. Cray et al (2011) proposed protein fraction measurements be done in plasma samples using gel electrophoresis instead of wet biochemistry analysis, since it presents less interferences. However, this methodology is more expensive than the one presented in this study and would be difficult to implement in most rehabilitation centers. Considering that blood biochemistry analysis is part of a set of health-evaluating data, the existence of possible over-or-underestimations in more than one measured analyte, due to pre-analytic factors, highly increases the chances of false diagnoses and re-introduction assessment mistakes. Thus, this study showed that the most indicated sample type for blood biochemistry evaluation is heparinized plasma. Despite its limitations, it is practical and produces reliable results, while minimizing sampling volume and material.

Figure 1
Image

Acknowledgements

This research was authorized by SISBIO 48654-1, www.icmbio.gov.br/sisbio, Authentication number 59739954.

References

Borboroglu, P. G., & Boersma, P. D. (Eds.). (2015). Penguins: natural history and conservation. University of Washington Press.

Cray, C., Wack, A., & Arheart, K. L. (2011). Invalid measurement of plasma albumin using bromcresol green methodology in penguins (Spheniscus species). Journal of avian medicine and surgery, 25(1), 14-22.

Hawkins, M. G.; Kass, P. H.; Zinkl, J. G.;Tell, L. A. (2006). Comparison of biochemical values in serum and plasma, fresh and frozen plasma, and hemolyzed samples from orange-winged Amazon parrots (Amazona amazonica). Veterinary Clinical Pathology, v. 35, n. 2, p. 219-225

Niemeyer, C., Vanstreels, R., Ruoppolo, V., Nascimento, T. A. D., Catão-Dias, J. L., Savioli, J., ... & Maracini, P. (2010). Resultados patológicos preliminares sobre a morte de pinguins de Magalhães (Spheniscus magellanicus) no litoral do estado de São Paulo. Anais da Semana Científica Benjamin Eurico Malucelli, 4(1), p-33.

Parsons, N. J., Schaefer, A. M., Van der Spuy, S. D., & Gous, T. A. (2015). Establishment of baseline haematology and biochemistry parameters in wild adult African penguins (Spheniscus demersus). Journal of the South African Veterinary Association, 86(1), 01-08.

Teare, J.A. (ed.): 2013, "Spheniscus_magellanicus_No_selection_by_gender__A ll_ages_combined_Conventional_American_units__2013 _CD.html" in ISIS Physiological Reference Intervals for Captive Wildlife: A CD-ROM Resource., International Species Information System, Eagan, MN.

Vanstreels, R. E. T.; Adornes A.C.; Cabana, A. L.; Niemeyer, C; Kolesnikovas, C.K.M.; Dantas, G.P.M.; Araújo, J.; Catão-Dias, J.L.;Groch, K.R.; Silva, L.A; Reisfeld, L. C.; Brandão,M. L.; Xavier, M.O.; Gonzalez -Vieira, O; Serafini, P. P.; Baldassin, P.; Canabarro, P. L.; Hurtado, R. F.; Silva-Filho, R.P.; Campos, S.D.E.; Ruoppolo, V. (2011). Manual de Campo para colheita e armazenamento de informações e amostras biológicas provenientes de pinguins-de-Magalhães (Spheniscus magellanicus). São Paulo: Centro Nacional de Pesquisa e Conservação de Aves Silvestres, 62 p.

Villouta, G., Hargreaves, R., & Rtveros, V. (1997). Haematological and clinical biochemistry findings in captive Humboldt penguins (Spheniscus humboldti). Avian Pathology, 26(4), 851-858.

Keywords: marine bird rescue, Health assessment, blood chemistry, Sample type, Anticoagulants

Conference: IMMR | International Meeting on Marine Research 2016, Peniche, Portugal, 14 Jul - 15 Jul, 2016.

Presentation Type: Poster presentation

Topic: Biodiversity, Conservation and Coastal Management

Citation: Morais JB, Volpato J, Costa A, Kolesnikovas C, De Souza M and Saito ME (2016). Improving the re-introduction of rescued Magellanic penguins through extended health evaluation with minimal sampling volume and material. Front. Mar. Sci. Conference Abstract: IMMR | International Meeting on Marine Research 2016. doi: 10.3389/conf.FMARS.2016.04.00089

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Received: 14 May 2016; Published Online: 13 Jul 2016.

* Correspondence: DVM. Julia B Morais, UDESC - Universidade do Estado de Santa Catarina, Veterinary Medicine, Lages, Santa Catarina, 88520-000, Brazil, juliebmorris@yahoo.com