SpringerLink
Log in

Effect of heparin and EDTA as anticoagulants on hematological values in farmed juvenile of Arapaima gigas

  • Published:
Aquaculture International Aims and scope Submit manuscript

Abstract

The rapid coagulation of fish blood makes the use of anticoagulants in hematological analysis necessary and means that choosing the most appropriate product is important. The aim of the present study was to evaluate the effect of two different anticoagulants (heparin sodium and K2EDTA), storage time, and temperature on the hematological values of Arapaima gigas juveniles. Twelve juveniles (905.79 ± 111.2 g and 49 ± 1.74 cm) kept in captivity were used for the tests. Blood was collected from the caudal vein and divided for the analysis of the effects of K2EDTA 10% and heparin sodium 5000 IU, storage time (0, 5, 10, and 24 h), and temperature (4 and 28 °C). The hematocrit, erythrocyte count, hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH), and differential leukocyte count were evaluated. The results revealed significant differences in the hematocrit, lymphocytes, and neutrophils when the anticoagulants were compared at 0 h of storage. In the hematocrit, heparin sodium produced variations, while K2EDTA maintained the values constant at 5 h, the erythrocyte count constant for up to 24 h, with the exception of at 4 °C, while hemoglobin produced variations in all the groups at 5 h. The blood indices MCV, MCHC, and MCH were best preserved, without variations, using K2EDTA at 28 °C for up to 5 h. In conclusion, K2EDTA 10% at 28 °C maintained constant hematological values of Arapaima gigas for 5 h, with the exception of hemoglobin, which should be analyzed as soon as possible.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Adeyemo OK, Okwilagwe OO, Ajani F (2009) Comparative assessment of sodium EDTA and heparin as anticoagulants for the evaluation of haematological parameters in cultured and feral African catfish (Clarias gariepinus). Braz J Aquat Sci Tech 13:19–24

    Article  CAS  Google Scholar 

  • Andrade JIA, Ono EA, Menezes GC, Brasil EM, Roubach R, Urbinati EC, Tavares-Dias M, Marcon JL, Affonso EG (2007) Influence of diets supplemented with vitamin C and E on pirarucu (Arapaima gigas) blood parameters. Comp Biochem Physiol Part A Mol Integr Physiol 146:576–580

    Article  Google Scholar 

  • Andrade-Porto SM, Affonso EG, Kochhann D, Malta JCO, Roque R, Ono EA, Araujo CSO, Tavares-Dias M (2017) Antiparasitic efficacy and blood effects of formalin on Arapaima gigas (Pisces: Arapaimidae). Aquaculture 479:38–44

    Article  CAS  Google Scholar 

  • Bain BJ, Lewis SM, Bates I (2006) Basic haematological techniques. In: Lewis SM, Bain BJ, Bates I (eds) Dacie and Lewis Practical Hematology, 10th edn. Churchill living stone Elsevier, Germany, pp 25–54

    Chapter  Google Scholar 

  • Bogdanova A, Makhro A, Wang J, Lipp P, Kaestner L (2013) Calcium in red blood cells: a perilous balance. Int J Mol Sci 4:9848–9872

    Article  Google Scholar 

  • Blaxhall PC (1972) The hematological assessment of the health of freshwater fish: a review of selected literature. J Fish Biol 4:593–604

    Article  CAS  Google Scholar 

  • Campbell TW (2015) Exotic animal hematology and cytology, 4th edn. Wiley-Blackwell, Ames

    Book  Google Scholar 

  • Campbell TW (2004) Hematology of Birds. In: Thrall MA (ed) Veterinary hematology and clinical chemistry. Lippincott Williams and Wilkins, New York, pp 225–258

    Google Scholar 

  • Cavero BAS, Ituassú DR, Pereira-Filho M, Roubach R, Bordinhon AM, da Fonseca FAL, Ono EA (2003) Uso de alimento vivo como dieta inicial no treinamento alimentar de juvenis de pirarucu. Pesq Agropec Bras 38:1011–1015

    Article  Google Scholar 

  • Ciepliński M, Kasprzak M, Grandtke M, Steliga A, Kamiński P, Jerzak L (2019) The effect of dipotassium EDTA and lithium heparin on hematologic values of farmed brown trout Salmo trutta (L.) spawners. Aquac Int 27:79–87

    Article  Google Scholar 

  • Dantas PDS, Chagas E, Dairiki J, Boi**k CDL (2012) Heparina e EDTA como anticoagulantes para pirarucu (Arapaima gigas). In: Morais RR, Boi**k CDL, Da Silva KE, Quisen RC (eds) Anais da IX Jornada de iniciação científica da Embrapa Amazônia Ocidental. Embrapa Amazônia Ocidental, Manaus, pp 203–209

    Google Scholar 

  • Dias MKR, Yoshioka ETO, Rodriguez AFR, Ribeiro RA, Faria FSEDV, Ozório ROA, Tavares-Dias M (2019) Growth, physiological and immune responses of Arapaima gigas (Arapaimidae) to Aeromonas hydrophila challenge and handling stress following feeding with immunostimulant supplemented diets. Fish Shellfish Immunol 84:843–847

    Article  Google Scholar 

  • Ekanem AP, Udoh AJ, Inyang-Etoh AP (2012) Effect of different anticoagulants on hematological parameters of Oreochromis niloticus. Int J Sci Adv Technol 2:17–20

    Google Scholar 

  • Faggio C, Arfuso F, Piccione G, Zumbo A, Fazio F (2014) Effect of three different anticoagulants and storage time on haematological parameters of Mugil cephalus (Linneaus, 1758). Turkish J Fish Aquat Sci 14:615–621

    Google Scholar 

  • Farias THV, Pereira NL, Pádua SBD, Alves LDO, Sakabe R, Belo MADA, Pilarski F (2016) Na2EDTA anticoagulant impaired blood samples from the teleost Piaractus mesopotamicus. Pesq Vet Bras 36:431–435

    Article  Google Scholar 

  • Fazio F (2019) Fish hematology analysis as an important tool of aquaculture: a review. Aquaculture 500:237–242

    Article  Google Scholar 

  • Fernandez-Méndez C, Troncoso AG, Green SB, Gonzales AF, Ruiz JM, Chirinos CR (2017) Artemia en la adaptación al consumo de alimento balanceado de paiche Arapaima gigas. Folia Amazón 26:187–194

    Article  Google Scholar 

  • Goldenfarb PB, Bowyer FP, Hall E, Brosious E (1971) Reproducibility in the hematology laboratory: the microhematocrit determination. Am J Clin Pathol 56:35–39

    Article  CAS  PubMed  Google Scholar 

  • Gonzales A, Mejía F, Huanuiri K, Sánchez I, Vásquez J, Fernández-Méndez C (2016) Valores hematológicos y bioquímicos de juveniles de paiche Arapaima gigas en cultivo intensivo. Folia Amazón 25:137–144

    Article  Google Scholar 

  • Harr KE, Raskin RE, Heard DJ (2005) Temporal effects of 3 commonly used anticoagulants on hematologic and biochemical variables in blood samples from macaws and Burmese pythons. Vet Clin Pathol 34:383–388

    Article  PubMed  Google Scholar 

  • Hattingh J (1975) Heparin and ethylenediamine tetra-acetate as anticoagulants for fish blood. Pflügers Archiv 355:347–352

    Article  CAS  PubMed  Google Scholar 

  • Hawkey CM, Dennett TB (1989) Color atlas of comparative veterinary hematology. Vet Clin Pathol 18:108–108

    Article  Google Scholar 

  • Imbiriba EP (2001) Production potential of pirarucu, Arapaima gigas, in captivity. Acta Amazon 31:299–299

    Article  Google Scholar 

  • Kaestner L (2013) Cation channels in erythrocytes - historical and future perspective. In: Calcium signalling. Springer, Berlin

    Chapter  Google Scholar 

  • Korcock DE, Houston AH, Gray JD (1988) Effects of sampling conditions on selected blood variables of rainbow trout, Salmo gairdneri Richardson. J Fish Biol 33:319–330

    Article  CAS  Google Scholar 

  • Knowles S, Hrubec TC, Smith SA, Bakal RS (2006) Hematology and plasma chemistry reference intervals for cultured shortnose sturgeon (Acipenser brevirostrum). Vet Clin Pathol 35:434–440

    Article  PubMed  Google Scholar 

  • Luz JR, Ramos APS, Melo JFB, Braga LGT (2019) Use of sodium butyrate in the feeding of Arapaima gigas (Schinz, 1822) juvenile. Aquaculture 510:248–255

    Article  CAS  Google Scholar 

  • Mafuvadze B, Erlwanger KH (2007) The effect of EDTA, heparin and storage on the erythrocyte osmotic fragility, plasma osmolality and haematocrit of adult ostriches (Struthio camelus). Vet Arh 77:427–434

    CAS  Google Scholar 

  • Maqbool A, Ahmed I, Sheikh ZA (2013) Effects of two commonly used anticoagulants on haematology and erythrocyte morphology of rainbow trout (Oncorhynchus mykiss). Int J Fish Aquat Stud 2:239–243

    Google Scholar 

  • Mirghaed AT, Ghelichpour M, Hoseini SM, Amini K (2017) Hemolysis interference in measuring fish plasma biochemical indicators. Fish Physiol Biochem 43:1143–1151

    Article  CAS  PubMed  Google Scholar 

  • Natt P, Herrick A (1952) A new blood diluent for counting the erythrocytes and leucocytes of the chicken. Poult Sci 31:735–738

    Article  Google Scholar 

  • Nuñez J (2009) Domestication de nouvelles espèces d'intérêt piscicole en Amazonie. Cah Agri 18:136–143

    Article  Google Scholar 

  • Oliveira EG, Pinheiro AB, de Oliveira VQ, da Silva-Júnior ARM, de Moraes MG, Rocha ÍRCB, Costa FHF (2012) Effects of stocking density on the performance of juvenile pirarucu (Arapaima gigas) in cages. Aquaculture 370:96–101

    Article  Google Scholar 

  • Oliveira AT, Pantoja-Lima J, Aride PHR, Tavares-Dias M, Marcon JL (2015) In: Tavares-Dias M, Mariano WS (eds) Fisiologia de arraias de água doce: subsídios para aplicabilidade na aquicultura. Aquicultura no Brasil: novas perspectivas, São Carlos, pp 45–74

    Google Scholar 

  • Pádua SBD, Pilarski F, Sakabe R, Dias-Neto J, Chagas EC, Ishikawa MM (2012) Heparin and K3EDTA as anticoagulants for tambaqui (Colossoma macropomum, Cuvier, 1816). Acta Amazon 42:293–298

    Article  Google Scholar 

  • Ranzani-Paiva MJ, Pádua SB, Tavares-Dias M, Egami M (2013) Métodos para análise hematológica em peixes. Eduem, Maringá

    Book  Google Scholar 

  • Sánchez I, Mejía F, Huanuiri K, Vásquez J, Gonzales A, Fernández-Méndez C (2017) Respuesta hematológica y bioquímica en juveniles de paiche Arapaima gigas sometidos a diferentes concentraciones de amonio. Folia Amazón 26:51–58

    Article  Google Scholar 

  • Smit GL, Hattingh J, Schoonbee HJ (1977) Observations on some effects of disodium ethylenediamine tetraacetate and heparin on fish blood. Comp Biochem Physiol C Toxicol Pharmacol 57:35–38

    Article  CAS  Google Scholar 

  • Tavares-Dias M, Sandrim EFS (1998) Influence of anticoagulants and blood storage on hematological values in tambaqui, Colossoma macropomum. Acta Sci Biol Sci 20:151–155

    Article  CAS  Google Scholar 

  • Tavares-Días M, De Moraes FR (2004) Hematologia de peixes teleósteos, 2nd edn. Ribeirão Preto, São Paulo

    Google Scholar 

  • Tavares Dias M, Moraes FR (2007) Haematological and biochemical reference intervals for farmed channel catfish. J Fish Biol 71:383–388

    Article  CAS  Google Scholar 

  • Walencik J, Witeska M (2007) The effects of anticoagulants on hematological indices and blood cell morphology of common carp (Cyprinus carpio L.). Comp Biochem Physiol C Toxicol Pharmacol 146:331–335

    Article  PubMed  Google Scholar 

  • Wintrobe M (1934) Variations on the size and haemoglobin content of erythrocytes in the blood various vertebrates. Folia Haematol 51:32–49

    Google Scholar 

  • Witeska M, Wargocka W (2011) Disodium EDTA used as anticoagulant causes hemolysis in common carp blood. Turkish J Vet Anim Sci 35:99–104

    Google Scholar 

  • Witeska M, Biardzka J, Kniaz J (2017) The effects of heparin concentration, storage time, and temperature on the values of hematological parameters in Cyprinus carpio. Turkish J Vet Anim Sci 41:351–356

    Article  CAS  Google Scholar 

  • Yousefi M, Abtahi B, Kenari AA (2012) Hematological, serum biochemical parameters, and physiological responses to acute stress of Beluga sturgeon (Huso huso, Linnaeus 1785) juveniles fed dietary nucleotide. Comp Clin Path 21:1043–1048

    Article  CAS  Google Scholar 

Download references

Data availability

All data generated or analyzed during this study are included in this published article.

Code availability

Not applicable.

Funding

The present study was funded by the project “Reproductive and nutritional study to increase the yield of aquaculture production in Loreto” (41-2016) of the Instituto de Investigaciones de la Amazonia Peruana (IIAP).

Author information

Authors and Affiliations

  1. Instituto de Investigaciones de la Amazonía Peruana (IIAP), Dirección de Investigación en Ecosistemas Acuáticos Amazónicos, Jr. Ica 1662, Puerto Maldonado, Perú

    Anai P. Gonzales-Flores

  2. Instituto de Investigaciones de la Amazonía Peruana (IIAP), Dirección de Investigación en Ecosistemas Acuáticos Amazónicos, Carretera Iquitos Nauta km 4.5, Iquitos, Perú

    Felix I. Mejia Perez, Katty A. Huanuiri Quinteros, Jorge L. Vasquez Rojas & Christian Fernandez-Mendez

  3. Departamento de Producción Animal, Facultad de Medicina Veterinaria y de Zootecnia, Universidad Nacional de Colombia, Bogotá, Colombia

    Ingrid Y. Sanchez Callejas

Authors
  1. Anai P. Gonzales-Flores
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Felix I. Mejia Perez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Katty A. Huanuiri Quinteros
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ingrid Y. Sanchez Callejas
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jorge L. Vasquez Rojas
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christian Fernandez-Mendez
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by APGF, FIMP, KAHQ, IYSC, and JLVR. The first draft of the manuscript was written by CFM, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Christian Fernandez-Mendez.

Ethics declarations

Ethics approval

Experimental procedures with animals were carried out in accordance with Peruvian legislation for animal protection and welfare in compliance with the protocol of the ethics committee for the use of animals in acts of experimentation, research, and teaching (30407/2016).

Conflict of interest

The authors declare no competing interests.

Additional information

Handling Editor: Dr. Gavin Burnell

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gonzales-Flores, A.P., Mejia Perez, F.I., Huanuiri Quinteros, K.A. et al. Effect of heparin and EDTA as anticoagulants on hematological values in farmed juvenile of Arapaima gigas. Aquacult Int 30, 263–271 (2022). https://doi.org/10.1007/s10499-021-00796-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10499-021-00796-1

Keywords

Search

Navigation