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Biomarker and hematological fieldwork with amphibians: is it necessary to sample all night?

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Abstract

In the context of the global amphibian crisis, biomonitoring constitutes a valuable assessment tool to provide critical up to date information on the status and health of amphibians worldwide. The aim of the current study was to evaluate the possible confounding effects of sex, size, and time since capture on enzymatic biomarkers and hematologic parameters of the South American frog Leptodactylus latrans. Frogs were collected by hand between 9 pm and 12 am on two consecutive nights. On the first night, captured frogs were transported for 2 h by car to laboratory installations, maintained overnight in plastic containers, and blood and tissue sampled on the next morning. In contrast, frogs collected on the second night were blood and tissue sampled in the field, immediately after the capture period. Hematological parameters were analyzed, and enzymatic activities of catalase, cholinesterase (ChE), and glutathione S-transferase (GST) were determined in the plasma, liver, kidney, and muscle. A sex difference was observed only for total white blood cell counts (WBC), females exhibiting significantly greater values than males. The packed cell volume (PCV), mean corpuscular hemoglobin concentration (MCHC), WBC, and muscle ChE activity were significantly correlated with snout-vent length (SVL). The correlation was inversed in the case of MCHC, WBC, and muscle ChE, while the correlation was positive between PCV and SVL. Most examined parameters presented similar values when frogs were sampled at night following capture or the next morning. Total red blood cells (RBCs) count, and plasma enzymatic activities of ChE and GST were the only parameters that presented significantly increased values in morning samplings compared with night samplings. Overall, the current study indicates that it is best to sample the frogs as soon as possible after capture if hematologic or plasmatic biomarkers are examined. Nevertheless, it is possible to sample on the next morning if tissular biomarkers are employed.

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References

  • Amiard-Triquet C, Berthet B (2015) Individual biomarkers. In: Amiard-Triquet C, Amiard J-C, Mouneyrac C (eds) Aquatic Ecotoxicology. Advancing Tools for Dealing with Emerging Risks. Academic Press, New York, USA

    Google Scholar 

  • Attademo AM, Peltzer PM, Lajmanovich RC, Cabagna M, Fiorenza G (2007) Plasma B-esterase and glutathione S-transferase activity in the toad Chaunus schneideri (Amphibia, Anura) inhabiting rice agroecosystems of Argentina. Ecotoxicology 16:533–539

    CAS  Google Scholar 

  • Attademo AM, Cabagna-Zenklusen M, Lajmanovich RC, Peltzer PM, Junges C, Basso A (2011) B-esterase activities and blood cell morphology in the frog Leptodactylus chaquensis (Amphibia: Leptodactylidae) on rice agroecosystems from Santa Fe Province (Argentina). Ecotoxicology 20:274–282

    CAS  Google Scholar 

  • Barnosky AD, Matzke N, Tomiya S, Wogan GOU, Swartz B, Quental TB, Marshall C, McGuire JL, Lindsey EL, Maguire KC, Mersey B, Ferrer EA (2011) Has the Earth’s sixth mass extinction already arrived? Nature 471:51–57

    CAS  Google Scholar 

  • Beauvais SL, Cole KJ, Atchison GJ, Coffey M (2002) Factors affecting brain cholinesterase activity in bluegill (Lepomis macrochirus). Water Air Soil Pollut 135:249–264

    CAS  Google Scholar 

  • Beckett GJ, Chapman BJ, Dyson EH, Hayes JD (1985) Plasma glutathione S-transferase measurements after paracetamol overdose: evidence for early hepatocellular damage. Gut 26:26–31

    CAS  Google Scholar 

  • Bennett MF, Harbottle JA (1968) The effects of hydrocortisone on the blood of tadpoles and frogs, Rana catesbeiana. Biol Bull 135:92–95

    CAS  Google Scholar 

  • Bennett MF, Gaudio CA, Johnson AO, Spisso JH (1972) Changes in the blood of newts, Notophthalmus viridescens, following administration of hydrocortisone. J Comp Physiol 80:233–237

    CAS  Google Scholar 

  • Bishop, P.J., Angulo, A., Lewis, J.P., Moore, R.D., Rabb, G.B., Garcia Moreno, J. 2012. The amphibian extinction crisis - what will it take to put the action into the amphibian conservation action plan? S.A.P.I.E.N.S [online], 5.2. http://sapiens.revues.org/140

  • Brodeur JC, Suarez RS, Natale GS, Ronco AE, Zaccagnini ME (2011) Reduced body condition and enzymatic alterations in frogs inhabiting intensive crop production areas. Ecotoxicol Environ Saf 74:1370–1380

    CAS  Google Scholar 

  • Brodeur, J.C., Vera Candioti, J. Soloneski, S. Larramendy, M.L., Ronco, A.E. 2012. Evidence of reduced feeding and oxidative stress in common tree frogs (Hypsiboas pulchellus) from an agroecosystem experiencing severe drought, J Herpetol 46:72–78

  • Brodeur JC, Vera Candioti J (2017) Impacts of agriculture and pesticides on amphibian terrestrial life stages: potential biomonitor/bioindicator species for the pampa region of Argentina. In: Larramendy ML (ed) Ecotoxicology and Genotoxicology - Non-traditional Terrestrial Models. Royal Society of Chemistry, London

    Google Scholar 

  • Brodeur JC, Sanchez M, Castro L, Rojas DE, Cristos D, Damonte MJ, Poliserpi MB, D’Andrea MF, Andriulo AE (2017) Accumulation of current-use pesticides, cholinesterase inhibition and reduced body condition in juvenile one-sided livebearer fish (Jenynsia multidentata) from the agricultural Pampa region of Argentina. Chemosphere 185:36–46

    CAS  Google Scholar 

  • Brodeur JC, Damonte MJ, Vera Candioti J, Poliserpi MB, D’Andrea MF, Bahl MF (2020) Frog body condition: basic assumptions, comparison of methods, and characterization of natural variability with field data from Leptodactylus latrans. Ecol Indic 112:106098

    Google Scholar 

  • Brousseau P, Pillet S, Frouin H, Auffret M, Gagne F, Fournier M (2013) Linking Immunotoxicity and Ecotoxicological effects at higher biological levels. In: Amiard-Triquet C, Amiard J-C, Rainbow PS (eds) Ecological Biomarkers. Indicators of Ecotoxicological Effects. CRC Press, New York

    Google Scholar 

  • Burton CB, Murray SA (1979) Effects of density on goldfish blood. J Hematology Comp Biochem Physiol 62A:555–558

    Google Scholar 

  • Cabagna MC, Lajmanovich RC, Stringhini G, Sanchez-Hernandez JC, Peltzer PM (2005) Hematological parameters of health status in the common toad Bufo arenarum in agroecosystems of Santa Fe Province. Argentina Applied Herpet 2:373–380

    Google Scholar 

  • Campbell Grant EH, Miller DAW, Schmidt BR, Adams MJ, Amburgey SM, Chambert T, Cruickshank SS, Fisher RN, Green DM, Hossack BR, Johnson PTJ, Joseph MB, Rittenhouse TAG, Ryan ME, Waddle JH, Walls SC, Bailey LL, Fellers GM, Gorman TA, Ray AM, Pilliod DS, Price SJ, Saenz D, Sadinski W, Muths E (2016) Quantitative evidence for the effects of multiple drivers on continental-scale amphibian declines. Sci Rep. https://doi.org/10.1038/srep25625

  • Camus L, Pampanin DM, Volpato E, Delaney E, Sanni S, Nasci C (2004) Total oxyradical scavenging capacity responses in Mytilus galloprovincialis transplanted into the Venice lagoon (Italy) to measure the biological impact of anthropogenic activities. Mar Pollut Bull 49:801–808

    CAS  Google Scholar 

  • Cathers T, Lewbart GA, Correa M, Stevens JB (1997) Serum chemistry and hematology values for anesthetized American bullfrogs (Rana catesbeiana). J Zoo Wildlife Med 28:171–174

    CAS  Google Scholar 

  • Cazenave J, Bacchetta C, Parma MJ, Scarabotti PA, Wunderlin DA (2009) Multiple biomarkers responses in Prochilodus lineatus allowed assessing changes in the water quality of Salado River basin (Santa Fe, Argentina). Environ Pollut 157:3025–3033

    CAS  Google Scholar 

  • Cei JM (1980) Amphibians of Argentina. Monitore zool ital Monogr 2:609 p

  • Christin SM, Ménard L, Giroux I, Marcogliese DJ, Ruby S, Cyr D, Fournier M, Brousseau P (2013) Effects of agricultural pesticides on the health of Rana pipiens frogs sampled from the field. Environ Sci Pollut Res 20:601–611

    CAS  Google Scholar 

  • Crump ML (2010) Amphibian diversity and life history. In: Dodd CK Jr (ed) Amphibian ecology and conservation. A handbook of techniques, Techniques in Ecology and Conservation Series. Oxford University Press, Oxford, U.K., pp 3–35

    Google Scholar 

  • Davies AK, Maney DL, Maerz JC (2008) The use of leukocyte profiles to measure stress in vertebrates: a review for ecologists. Funct Ecol 22:760–772

    Google Scholar 

  • Davies AK, Durso AM (2009) White blood cell differentials of northern cricket frogs (Acris c. crepitans) with a compilation of published values from other amphibians. Herpetologica 65(3):260–267

    Google Scholar 

  • Drabkin DL, Austin JH (1935) Spectrophotometric studies. II. Preparations from washed blood cells; nitric oxide hemoglobin and sulfhemoglobin. J Biol Chem 112:51–65

    CAS  Google Scholar 

  • Ellman GL, Courtney KD, Valentino AJ, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95

  • Falfushinska H, Loumbourdis N, Romanchuk L, Stolyar O (2008) Validation of oxidative stress responses in two populations of frogs from Western Ukraine. Chemosphere 73(7):1096–1101

    CAS  Google Scholar 

  • Fernandez G, Mena MP, Arnau A, Sanchez O, Soley M, Ramirez I (2000) Immobilization stress induces c-Fos accumulation in liver. Cell Stress Chaperones 5(4):306–312

    CAS  Google Scholar 

  • Flammarion P, Noury P, Garric J (2002) The measurement of cholinesterase activities as a biomarker in chub (Leuciscus cephalus): the fish length should not be ignored. Environ Pollut 120(2):325–330

    CAS  Google Scholar 

  • Forbes MR, McRuer DL, Shutler D (2006) White blood cell profiles of breeding American toads (Bufo americanus) relative to sex and body size. Comp Clin Pathol 15:155–159

    Google Scholar 

  • Forzán MJ, Heatley J, Russell KE, Horney B (2017) Clinical pathology of amphibians: a review. Vet Clin Pathol 46(1):11–33

    Google Scholar 

  • George ADI, Akinrotimi OA (2017) Influence of sex on haematological response of Clarias gariepinus juveniles treated with atrazine and metalochlor. Trends Green Chem 3:1–6

    Google Scholar 

  • Habig C, Di Giulio RT (1991) Biochemical characteristics of cholinesterases in aquatic organisms. In: Mineau P (ed) Cholinesterase-inhibiting insecticides. Elsevier, Amsterdam, pp 20–33

    Google Scholar 

  • Hansen P-D (2003) Biomarkers. In: Markert BA, Breure AM, Zechmeister HG (eds) Bioindicators and biomonitors. Elsevier Science Ltd, Oxford

    Google Scholar 

  • Heatley JJ, Johnson M (2009) Clinical technique: amphibian hematology: a practitioner’s guide. J Exotic Pet Med 18:14–19

    Google Scholar 

  • Hegde G, Krishnamurthy SV (2014) Analysis of health status of the frog Fejervarya limnocharis (Anura: Ranidae) living in rice paddy fields of Western Ghats, using body condition factor and AChE content. Ecotoxicol Environ Contam 9:69–76

    Google Scholar 

  • Heyer, R., Langone, J., La Marca, E., Acevedo-Ramos, C., di Tada, I., Baldo, D., Lavilla, E., Scott, N., Aquino, L., Hardy, J. 2010. Leptodactylus latrans. The IUCN red list of threatened species: e.T57151A11592655

  • Huggett RJ, Kimerle RA, Mehrle PM, Bergman HL (2018) Biomarkers: biochemical, physiological, and histological markers of anthropogenic stress. CRC Press, Boca Raton

    Google Scholar 

  • Imperato A, Puglisi-Allegra S, Casolini P, Angellucci L (1991) Changes in brain dopamine and acetylcholine release during and following stress are independent of the pituitary-adrenocortical axis. Brain Res 538:111–117

    CAS  Google Scholar 

  • IPBES. 2019. Global assessment report on biodiversity and ecosystem services of the intergovernmental science-policy platform on biodiversity and ecosystem services. E.S. Brondizio, J. Settele, S. Díaz, and H.T. Ngo (editors). IPBES secretariat, Bonn, Germany

  • Kaufer D, Friedman A, Seidman S, Soreq H (1998) Acute stress facilitates long-lasting changes in cholinergic gene expression. Nature 393:373–377

    CAS  Google Scholar 

  • Koenig S, Sole M (2013) Muscular cholinesterase and lactate dehydrogenase activities in deep-sea fish from the NW Mediterranean. Mar Environ Res 94:16–23

    Google Scholar 

  • Lajmanovich RC, Sanchez-Hernandez JC, Stringhini G, Peltzer PM (2004) Levels of serum cholinesterase activity in the rococo toad (Bufo paracnemis) in agrosystems of Argentina. Bull Environ Contam Toxicol 72(3):586–591

    CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–75

  • Mahapatra BB, Das M, Dutta SK, Mahapatra PK (2012) Hematology of Indian rhacophorid tree frog Polypedates maculatus gray, 1833 (Anura: Rhacophoridae). Comp Clin Pathol 21:453–460

    Google Scholar 

  • Maneyro R, Naya DE, da Rosa I, Canavero A, Camargo A (2004) Diet of the south American frog Leptodactylus ocellatus (Anura, Leptodactylidae) in Uruguay, Iheringia, Sér. Zool. Porto Alegre 94:57–61

    Google Scholar 

  • Menezes S, Soares AMVM, Guilhermino L, Peck MR (2006) Biomarker responses of the estuarine brown shrimp Crangon crangon L. to non-toxic stressors: temperature, salinity and handling stress effects. J Exp Mar Biol Ecol 335:114–122

    Google Scholar 

  • Meshorer E, Erb C, Gazit R, Pavlovsky L, Kaufer D, Friedman A, Glick D, Ben-Arie N, Soreq H (2002) Alternative splicing and neuritic mRNA translocation under long-term neuronal hypersensitivity. Science 295:508–512

    CAS  Google Scholar 

  • Meesawat, S., Kitana, N., Kitana, J. 2016. Hematology of wild caught Hoplobatrachus rugulosus in northern Thailand. Asian Herpetol. Res. 7(2):131–138

  • Mitchell MA (2009) Anesthetic considerations for amphibians. J Exotic Pet Med 18:40–49

    Google Scholar 

  • Monaghan P, Spencer KA (2014) Stress and life history. Curr Biol 24(10):R408–R412

    CAS  Google Scholar 

  • Motlagh SP, Zarejabad AM, Nasrabadi RG, Ahmadifar E, Molaee M (2012) Haematology, morphology and blood cells characteristics of male and female Siamese fighting fish (Betta splendens). Comp Clin Pathol 21(1):15–21

    Google Scholar 

  • Moura Costa DD, Filipak, Neto F, Costa MDM, Morais RN, Garcia JRE, Esquivel BM, Ribeiro O, C.A. (2010) Vitellogenesis and other physiological responses induced by 17-β-estradiol in males of freshwater fish Rhamdia quelen. Comp Biochem Physiol 151C:248–257

    Google Scholar 

  • Narayan E, Hero JM (2011) Urinary corticosterone responses and haematological stress indicators in the endangered Fijian ground frog (Platymantis vitiana) during transportation and captivity. Aust J Zool 59:79–85

    Google Scholar 

  • Narayan EJ, Cockrem JF, Hero JM (2013) Sight of a predator induces a corticosterone stress response and generates fear in an amphibian. PLoS One 8(8):e73564. https://doi.org/10.1371/journal.pone.0073564

    Article  CAS  Google Scholar 

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

    Google Scholar 

  • Omeje VO, Lambrechts H, Brink D (2019) Hepatic and haemato-biochemical alterations in juvenile Mozambique tilapia (Oreochromis mossambicus) on pawpaw (Carica papaya) seed meal. J Agric Sci 11(8):20–30

    Google Scholar 

  • Orton F, Baynes A, Clare F, Duffus ALJ, Larroze S, Scholze M, Garner TWJ (2014) Body size, nuptial pad size and hormone levels: potential non-destructive biomarkers of reproductive health in wild toads (Bufo bufo). Ecotoxicology 23(7):1359–1365

    CAS  Google Scholar 

  • Pathiratne A, Chandrasekera WHU, De Seram PKC (2008) Effects of biological and technical factors on brain and muscle cholinesterases in Nile tilapia, Oreochromis niloticus: implications for biomonitoring neurotoxic contaminations. Arch Environ Contam Toxicol 54:309–3017

    CAS  Google Scholar 

  • Parida S, Panga R, Rajappa M, Kundra P (2018) Study of glutathione S-transferase levels in patients receiving intravenous paracetamol perioperatively: a randomized controlled trial. Indian J Gastroenterol 37(6):511–519

    Google Scholar 

  • Pimm SL, Jenkins CN, Abell R, Brooks TM, Gittleman JL, Joppa LN, Raven PH, Roberts CM, Sexton JO (2014) The biodiversity of species and their rates of extinction, distribution, and production. Science 344:987–998

    CAS  Google Scholar 

  • Peig J, Green AJ (2009) New perspectives for estimating body condition from mass/length data: the scaled mass index as an alternative. Oikos 118:1883–1891

    Google Scholar 

  • Phillips TA, Summerfelt RC, Atchison GJ (2002) Environmental, biological, and methodological factors affecting cholinesterase activity in walleye (Stizostedion vitreum). Arch Environ Contam Toxicol 43(1):75–80

    CAS  Google Scholar 

  • Pollo FE, Grenat PR, Salinas ZA, Otero MA, Salas NE, Martino AL (2017) Evaluation in situ of genotoxicity and stress in South American common toad Rhinella arenarum in environments related to fluorite mine. Environ Sci Pollut Res 24(22):18179–18187.

  • Pounds, J.A., Bustamante, M.R., Coloma, L.A., Consuegra, J.A, Fogden, M.P.L., Foster, P.N., La Marca, E., Masters, K.L., Merino-Viteri, A., Puschendorf, R., et al. 2006. Widespread amphibian extinctions from epidemic disease driven by global warming. Nature 439:161–167

  • Prokic MD, Petrovic TG, Gavric JP, Despotovic SG, Gavrilovic BR, Radovanovic TB, Faggio C, Saicic ZS (2018) Comparative assessment of the antioxidative defense system in subadult and adult anurans: a lesson from the Bufotes viridis toad. Zoology 130:30–37

    Google Scholar 

  • Roelants K, Gower DJ, Wilkinson M, Loader SP, Biju SD, Guillaume K, Moriau L, Bossuyt F (2007) Global patterns of diversification in the history of modern amphibians. Proc Natl Acad Sci U S A 104:887–892

    CAS  Google Scholar 

  • Santymire RM, Manjerovic MB, Sacertote-Velat A (2018) A novel method for the measurement of glucocorticoids in dermal secretions of amphibians. Conserv Physiol 6(1):coy008. https://doi.org/10.1093/conphys/coy008

    Article  CAS  Google Scholar 

  • Stuart SN, Chanson JS, Cox NA, Young BE, Rodrigues ASL, Fischman DL, Waller RW (2004) Status and trends of amphibian declines and extinctions worldwide. Science 306:1783–1785

    CAS  Google Scholar 

  • Tecles F, Escribano D, Martinez-Miro S, Hernandez F, Contreras MD, Ceron JJ (2016) Cholinesterase in porcine saliva: analytical characterization and behavior after experimental stress. Res Vet Sci 106:23–28

    CAS  Google Scholar 

  • Tsakiris S, Kontopoulos AN (1993) Time changes in Na +, K+-ATPase, mg+-ATPase, and acetylcholinesterase activities in the rat cerebrum and cerebellum caused by stress. Pharmacol Biochem Behav 44:339–342

    CAS  Google Scholar 

  • Van der Oost R, Beyer J, Vermeulen NP (2003) Fish bioaccumulation and biomarkers in environmental risk assessment: a review. Environ Toxicol Pharmacol 13:57–149

    Google Scholar 

  • Voorhees JL, Powell ND, Moldovan L, Mo X, Eubank TD, Clay BM (2013) Chronic restraint stress upregulates erythropoiesis through glucocorticoid stimulation. PLoS One 8(10):e77935. https://doi.org/10.1371/journal.pone.0077935

    Article  CAS  Google Scholar 

  • Wake DB, Vredenburg VT (2008) Are we in the midst of the sixth mass extinction? A view from the world of amphibians. Proc Natl Acad Sci U S A 105:11466–11473

    CAS  Google Scholar 

  • Williams M, Zalasiewicz J, Haff PK, Schwägerl C, Barnosky AD, Ellis EC (2015) The Anthropocene biosphere. The Anthropocene Rev 2:196–219

    Google Scholar 

  • Wilson JT, Pascoe PL, Parry JM, Dixon DR (1998) Evaluation of the comet assay as a method for the detection of DNA damage in the cells of a marine invertebrate, Mytilus edulis L. (Mollusca: Pelecypoda). Mutat Res 399:87–95

    CAS  Google Scholar 

  • Xiong J, Zhang Y, Chen W, Min Y, Gou J (2018) Some haematological parameters of wild caught warty toothed toad Oreolalax rugosus (Liu, 1943) (Anura: Megophryidae). Acta Zool Bulg 70(1):69–74

    Google Scholar 

  • Young S, Warner J, Speare R, Berger L, Skerratt LF, Muller R (2012) Hematologic and plasma biochemical reference intervals for health monitoring of wild Australian tree frogs. Vet Clin Pathol 41(4):478–492

    Google Scholar 

  • Zhelev ZM, Georgieva KN, Todorov OB, Peeva KG (2017) Haematological parameters of Bufotes viridis (Laurenti, 1768) (Anura: Bufonidae) from southern Bulgaria. Acta Zool Bulg 69(3):335–343

    Google Scholar 

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Funding

Funding for this work was provided by the “Agencia Nacional de Promoción Científica y Técnica” and the “Instituto Nacional de Tecnología Agropecuaria”.

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  1. Instituto de Recursos Biológicos, Centro de Investigaciones de Recursos Naturales (CIRN), Centro Nacional de Investigaciones Agropecuarias (CNIA), Instituto Nacional de Tecnología Agropecuaria (INTA), 1686, Hurlingham, Buenos Aires, Argentina

    Julie Céline Brodeur & María Belén Poliserpi

  2. Centro Investigaciones del Medio Ambiente, Departamento de Química, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina

    María Florencia Bahl & Guillermo Sebastián Natale

  3. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina

    Julie Céline Brodeur, María Florencia Bahl & Guillermo Sebastián Natale

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  1. Julie Céline Brodeur
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Brodeur, J.C., Bahl, M.F., Natale, G.S. et al. Biomarker and hematological fieldwork with amphibians: is it necessary to sample all night?. Environ Sci Pollut Res 27, 17152–17161 (2020). https://doi.org/10.1007/s11356-020-08313-2

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