Abstract
Physiological tradeoffs occur in organisms coping with their environments, which are likely to increase as populations reach peripheries of established ranges. Invasive species offer opportunities to study tradeoffs that occur, with many hypotheses focusing on how immune responses vary during dispersal. The cane toad (Rhinella marina) is a well-known invasive species. Populations near the expanding edge of the Australian invasion have altered immune responses compared to toads from longer-established core populations, although this has not been well-documented for Florida populations. In this study, cane toads from a northern edge [New Port Richey (NPR)] and southern core (Miami) population in Florida were collected and injected with lipopolysaccharide (LPS) to compare immune responses. Core population individuals injected with LPS showed greater metabolic increases compared to their baseline rates that were higher compared to those from the edge population. In addition, LPS-injected core individuals had different circulating leukocyte profiles compared to saline-injected cane toads while edge individuals did not. There was a significant interaction between plasma bacteria-killing capability (BKA) and treatment, such that BKA decreased with time in saline compared to LPS-injected individuals, and saline-injected toads from the edge population had lower BKA compared to LPS-injected edge toads at 20 h post-injection. There was also a significant interaction between location and time on circulating corticosterone (CORT) levels following injections with saline or LPS, with CORT decreasing more with time in core population toads. The differential CORT response indicates that differential stress responses contribute to the tradeoffs observed with immunity and dispersal.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data accessibility
The data supporting the results will be archived in Mendeley Data: https://doi.org/10.17632/hk24s8jrhk.1
Abbreviations
- NPR:
-
New Port Richey
- LPS:
-
Lipopolysaccharide
- BKA:
-
Bacteria killing assay
- GC:
-
Glucocorticoid
- CORT:
-
Corticosterone
- N:L:
-
Neutrophil:Lymphocyte
- V̇O2 :
-
Volume (ml) of oxygen consumed h−1
References
Andino FDJ, Chen G, Li Z, Grayfer L, Robert J (2012) Susceptibility of Xenopus laevis tadpoles to infection by the ranavirus Frog-Virus 3 correlates with a reduced and delayed innate immune response in comparison with adult frogs. Virology 432:435–443. https://doi.org/10.1016/j.virol.2012.07.001
Assis VR, Titon SCM, Barsotti AMG, Spira B, Gomes FR (2013) Antimicrobial capacity of plasma from anurans of the Atlantic Forest. S Am J Herpetol 8:155–161. https://doi.org/10.2994/SAJH-D-13-00007.1
Bennett MF, Alspaugh JK (1964) Some changes in the blood of frogs following administration of hydrocortisone. Va J Sci 15:76–79
Bertrand S, Criscuolo F, Faivre B, Sorci G (2006) Immune activation increases susceptibility to oxidative tissue damage in zebra finches. Funct Ecol 20:1022–1027. https://doi.org/10.1111/j.1365-2435.2006.01191.x
Bickel M (1993) The role of interleukin-8 in inflammation and mechanisms of regulation. J Periodontol 64:456–460
Bishop CR, Athens JW, Boggs DR, Warner HR, Cartwright GE, Wintrobe MM (1968) Leukokinetic studies: XIII. A non-steady-state kinetic evaluation of the mechanism of cortisone-induced granulocytosis. J Clin Investig 47:249–260
Bonier F, Martin PR, Moore IT, Wingfield JC (2009) Do baseline glucocorticoids predict fitness? Trends Ecol Evol 24:634–642. https://doi.org/10.1016/j.tree.2009.04.013
Brown GP, Shilton CM, Shine R (2011) Measuring amphibian immunocompetence: validation of the phytohemagglutinin skin-swelling assay in the cane toad, Rhinella marina. Methods Ecol Evol 2:341–348. https://doi.org/10.1111/j.2041-210X.2011.00090.x
Brown GP, Shine R (2014) Immune response varies with rate of dispersal in invasive cane toads (Rhinella marina). PLoS ONE 9:e99734. https://doi.org/10.1371/journal.pone.0099734
Brown GP, Kelehear C, Shilton CM, Phillips BL, Shine R (2015a) Stress and immunity at the invasion front: a comparison across cane toad (Rhinella marina) populations. Biol J Linn Soc 116:748–760. https://doi.org/10.1111/bij.12623
Brown GP, Phillips BL, Dubey S, Shine R (2015b) Invader immunology: invasion history alters immune system function in cane toads (Rhinella marina) in tropical Australia. Ecol Lett 18:57–65. https://doi.org/10.1111/ele.12390
Careau V, Thomas D, Humphries MM, Réale D (2008) Energy metabolism and animal personality. Oikos 117:641–653. https://doi.org/10.1111/j.0030-1299.2008.16513.x
Castell JV, Gomez-Lechon MJ, David M, Andus T, Geiger T, Trullenque R, Heinrich PC (1989) Interleukin-6 is the major regulator of acute phase protein synthesis in adult human hepatocytes. FEBS Lett 242:237–239
Davis 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. https://doi.org/10.1111/j.1365-2435.2008.01467.x
Dhabhar FS (2002) Stress-induced augmentation of immune function—the role of stress hormones, leukocyte trafficking, and cytokines. Brain Behav Immun 16:785–798. https://doi.org/10.1016/S0889-1591(02)00036-3
Dierich MP, Bitter-Suermann D, König W, Hadding U, Galanos C, Rietschel ET (1973) Analysis of bypass activation of C3 by endotoxic LPS and loss of this potency. Immunology 24(4):721
Dinarello CA (2009) Immunological and inflammatory functions of the Interleukin-1 family. Annu Rev Immunol 27:519–550. https://doi.org/10.1146/annurev.immunol.021908.132612
Dupoué A, Brischoux F, Lourdais O, Angelier F (2013) Influence of temperature on the corticosterone stress–response: an experiment in the Children’s python (Antaresia childreni). Gen Comp Endocrinol 193:178–184. https://doi.org/10.1016/j.ygcen.2013.08.004
Eckert CG, Samis KE, Lougheed SC (2008) Genetic variation across species’ geographical ranges: the central-marginal hypothesis and beyond. Mol Ecol 17:1170–1188. https://doi.org/10.1111/j.1365-294X.2007.03659.x
Eikenaar C, Husak J, Escallon C, Moore IT (2012) Variation in testosterone and corticosterone in amphibians and reptiles: relationships with latitude, elevation, and breeding season length. Am Nat 180:642–654. https://doi.org/10.1086/667891
Evans MR, Roberts ML, Buchanan KL, Goldsmith AR (2006) Heritability of corticosterone response and changes in life history traits during selection in the zebra finch. J Evol Biol 19:343–352. https://doi.org/10.1111/j.1420-9101.2005.01034.x
Galanos C, Rietschel ET, Lüderitz O, Westphal O (1971) Interaction of lipopolysaccharides and lipid A with complement. Eur J Biochem 19:143–152
Gardner S, Assis VR, Zhao H, Gomes FR, Peatman E, Mendonça MT (2018) Differential gene expression to an LPS challenge in relation to exogenous corticosterone in the invasive cane toad (Rhinella marina). Dev Comp Immunol 88:114–123. https://doi.org/10.1016/j.dci.2018.07.016
Gervasi SS, Hunt EG, Lowry M, Blaustein AR (2014) Temporal patterns in immunity, infection load and disease susceptibility: understanding the drivers of host responses in the amphibian-chytrid fungus system. Funct Ecol 28:569–578. https://doi.org/10.1111/1365-2435.12194
Gewurz H, Wernick PR, Quie PG, Good RA (1965) Effects of hydrocortisone succinate on the complement system. Nature 208:755–757
Goessling JM, Guyer C, Mendonça MT (2017) More than fever: thermoregulatory responses to immunological stimulation and consequences of thermoregulatory strategy on innate immunity in Gopher Tortoises (Gopherus polyphemus). Physiol Biochem Zool 90:484–493. https://doi.org/10.1086/692116
Goetz SM, Romagosa CM, Appel AG, Guyer C, Mendonça MT (2017) Reduced innate immunity of Cuban Treefrogs at leading edge of range expansion. J Exp Zool 327:592–599. https://doi.org/10.1002/jez.2146
Gomes FR, Oliveira RV, Assis VR, Junior BT, Moretti EH, Mendonça MT (2012) Interspecific variation in innate immune defenses and stress response of toads from Botucatu (São Paulo, Brazil). South Am J Herpetol 7:1–9. https://doi.org/10.2994/057.007.0101
Götze O, Müller-Eberhard HJ (1971) The C3-activator system: an alternate pathway of complement activation. J Exp Med 134(3):90–108
Graham SP, Kelehear C, Brown GP, Shine R (2012) Corticosterone–immune interactions during captive stress in invading Australian cane toads (Rhinella marina). Horm Behav 62:146–153. https://doi.org/10.1016/j.yhbeh.2012.06.001
Greenspan SE, Bower DS, Webb RJ, Berger L, Rudd D, Schwarzkopf L, Alford RA (2017) White blood cell profiles in amphibians help to explain disease susceptibility following temperature shifts. Dev Comp Immunol 77:280–286. https://doi.org/10.1016/j.dci.2017.08.018
Hayward LS, Wingfield JC (2004) Maternal corticosterone is transferred to avian yolk and may alter offspring growth and adult phenotype. Gen Comp Endocrinol 135:365–371. https://doi.org/10.1016/j.ygcen.2003.11.002
Hu F, Crespi EJ, Denver RJ (2008) Programming neuroendocrine stress axis activity by exposure to glucocorticoids during postembryonic development of the frog, Xenopus laevis. Endocrinology 149:5470–5481. https://doi.org/10.1210/en.2008-0767
Husak JF, Moore IT (2008) Stress hormones and mate choice. Trends Ecol Evol 23:532–534. https://doi.org/10.1016/j.tree.2008.06.007
Jennings JF, Taylor G (1964) Effect of hydrocortisone hemi-succinate on immune lysis of sheep erythrocytes. Nature 203:661
Kitaysky AS, Kitaiskaia EV, Piatt JF, Wingfield JC (2005) A mechanistic link between chick diet and decline in seabirds? Proc Royal Soc Lond 273:445–450. https://doi.org/10.1098/rspb.2005.3351
Koolhaas JM, Korte SM, De Boer SF, Van Der Vegt BJ, Van Reenen CG, Hopster H, De Jong IC, Ruis MA, Blokhuis HJ (1999) Coping styles in animals: current status in behavior and stress-physiology. Neurosci Biobehav Rev 23:925–935. https://doi.org/10.1016/S0149-7634(99)00026-3
Krakauer T (1968) The ecology of the neotropical toad, Bufo marinus, in south Florida. Herpetologica 24:214–221
Lee KA, Klasing KC (2004) A role for immunology in invasion biology. Trends Ecol Evol 19:523–529. https://doi.org/10.1016/j.tree.2004.07.012
Lee KA, Martin LB, Wikelski MC (2005) Responding to inflammatory challenges is less costly for a successful avian invader, the house sparrow (Passer domesticus), than its less-invasive congener. Oecologia 145:243–250. https://doi.org/10.1007/s00442-005-0113-5
Lever C (2001) The Cane toad: the history and ecology of a successful colonist. Westbury Scientific Publishing, Otley
Llewellyn D, Brown GP, Thompson MB, Shine R (2010) Behavioral responses to immune-system activation in an anuran (the cane toad, Bufo marinus): field and laboratory studies. Physiol Biochem Zool 84:77–86. https://doi.org/10.1086/657609
Llewellyn D, Thompson MB, Brown GP, Phillips BL, Shine R (2012) Reduced investment in immune function in invasion-front populations of the cane toad (Rhinella marina) in Australia. Biol Invasions 14:999–1008. https://doi.org/10.1007/s10530-011-0135-3
Lighton JR (2018) Measuring metabolic rates: a manual for scientists. Oxford University Press, Oxford
Loos M, Bitter-Suermann D, Dierich M (1974) Interaction of the first (C1), the second (C2) and the fourth (C4) component of complement with different preparations of bacterial lipopolysaccharides and with lipid A. J Immunol 112:935–940
Mahony SM, Tisdale MJ (1990) Metabolic effects of tumour necrosis factor alpha in NMRI mice. Br J Cancer 61:514–519
Martin LB, Kidd L, Liebl AL, Coon CA (2011) Captivity induces hyper-inflammation in the house sparrow (Passer domesticus). J Exp Biol 214:2579–2585. https://doi.org/10.1242/jeb.057216
Mendonça MT, Chernetsby SD, Nester KE, Gardner GL (1996) Effects of sex steroids on sexual behavior in the big brown bat, Eptesicus fuscus. Horm Behav 30:153–161. https://doi.org/10.1006/hbeh.1996.0019
Mittan CS, Zamudio KR (2019) Rapid adaptation to cold in the invasive cane toad Rhinella marina. Conserv Physiol 7:1–12. https://doi.org/10.1093/conphys/coy075
Morrison DC, Kline LF (1977) Activation of the classical and properdin pathways of complement by bacterial lipopolysaccharides (LPS). J Immunol 118:362–368
Nemeth E, Rivera S, Gabayan V, Keller C, Taudorf S, Pedersen BK, Ganz T (2004) IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin. J Clin Invest 113:1271–1276
Norris K, Evans MR (2000) Ecological immunology: life history trade-offs and immune defense in birds. Behav Ecol 11:19–26. https://doi.org/10.1093/beheco/11.1.19
Øverli Ø, Sørensen C, Pulman KG, Pottinger TG, Korzan W, Summers CH, Nilsson GE (2007) Evolutionary background for stress-coping styles: relationships between physiological, behavioral, and cognitive traits in non-mammalian vertebrates. Neurosci Biobehav Rev 31:396–412. https://doi.org/10.1016/j.neubiorev.2006.10.006
Packard BD, Weiler JM (1983) Steroids inhibit activation of the alternative-amplification pathway of complement. Infect Immun 40:1011–1014
Padgett DA, Glaser R (2003) How stress influences the immune response. Trends Immunol 24(8):444–448. https://doi.org/10.1016/S1471-4906(03)00173-X
Peterson JD, Steffen JE, Reinert LK, Cobine PA, Appel A, Rollins-Smith L, Mendonça MT (2013) Host stress response is important for the pathogenesis of the deadly amphibian disease, chytridiomycosis, in Litoria caerulea. PLoS ONE 8:e62146. https://doi.org/10.1371/journal.pone.0062146
Phillips BL, Kelehear C, Pizzatto L, Brown GP, Barton D, Shine R (2010) Parasites and pathogens lag behind their host during periods of host range advance. Ecology 91:872–881. https://doi.org/10.1890/09-0530.1
Punzo F, Lindstrom L (2001) The toxicity of eggs of the giant toad, Bufo marinus to aquatic predators in a Florida retention pond. J Herpetol 35:693–697. https://doi.org/10.2307/1565916
Rollins-Smith LA (2001) Neuroendocrine-immune system interactions in amphibians: implications for understanding global amphibian declines. Immunol Res 23(2–3):273–280
Rollins-Smith LA (2017) Amphibian immunity–stress, disease, and climate change. Dev Comp Immunol 66:111–119. https://doi.org/10.1016/j.dci.2016.07.002
Sagarin RD, Gaines SD (2002) The abundant centre distribution: to what extent is it a biogeographical rule? Ecol Lett 5:137–147. https://doi.org/10.1046/j.1461-0248.2002.00297.x
Scalf CS, Chariker JH, Rouchka EC, Ashley NT (2019) Transcriptomic analysis of immune response to bacterial lipopolysaccharide in zebra finch (Taeniopygia guttata). BMC Genomics 20:647. https://doi.org/10.1186/s12864-019-6016-3
Selechnik D, West AJ, Brown GP, Fanson KV, Addison B, Rollins LA, Shine R (2017) Effects of invasion history on physiological responses to immune system activation in invasive Australian cane toads. PeerJ 5:e3856. https://doi.org/10.7717/peerj.3856
Sheldon BC, Verhulst S (1996) Ecological immunology: costly parasite defences and trade-offs in evolutionary ecology. Trends Ecol Evol 11:317–321. https://doi.org/10.1016/0169-5347(96)10039-2
Sherman E, Stephens A (1998) Fever and metabolic rate in the toad Bufo marinus. J Therm Biol 1:49–52. https://doi.org/10.1016/S0306-4565(97)00045-4
Shine R (2010) The ecological impact of invasive cane toads (Bufo marinus) in Australia. Q Rev Biol 85:253–291. https://doi.org/10.1086/655116
Smith GD, Neuman-Lee LA, Webb AC, Angilletta MJ, DeNardo DF, French SS (2017) Metabolic responses to different immune challenges and varying resource availability in the side-blotched lizard (Uta stansburiana). J Comp Physiol B 187:1173–1182. https://doi.org/10.1007/s00360-017-1095-4
Stouthard JM, Romijn JA, Van der Poll T, Endert EK, Klein S, Bakker PJ et al (1995) Endocrinologic and metabolic effects of interleukin-6 in humans. Am J Physiol Endocrinol Metab 268:E813–E819
Tredget EE, Yu YM, Zhong S, BuriniOkusawa RS, Gelfand JA et al (1988) Role of interleukin 1 and tumor necrosis factor on energy metabolism in rabbits. Am J Physiol Endocrinol Metab 255:E760–E768
Trumbo DR, Epstein B, Hohenlohe PA, Alford RA, Schwarzkopf L, Storfer A (2016) Mixed population genomics support for the central marginal hypothesis across the invasive range of the cane toad (Rhinella marina) in Australia. Mol Ecol 25:4161–4176. https://doi.org/10.1111/mec.13754
Tuomi J, Hakala T, Haukioja E (1983) Alternative concepts of reproductive effort, costs of reproduction, and selection in life-history evolution. Am Zool 23:25–34. https://doi.org/10.1093/icb/23.1.25
U.S. Geological Survey (2020) Specimen observation data for Rhinella marina (Linnaeus, 1758), Nonindigenous Aquatic Species Database, Gainesville, FL, ttps://nas.er.usgs.gov/viewer/omap.aspx?SpeciesID=48. Accessed 13 Jan 2020
Van Der Poll T, Romijn JA, Endert EK, Borm JJ, Buller HR, Sauerwein HP (1991) Tumor necrosis factor mimics the metabolic response to acute infection in healthy humans. Am J Physiol Endocrinol Metab 261:E457–E465
Varela M, Romero A, Dios S, Van der Vaart M, Figueras A, Meijer AH, Novoa B (2014) Cellular visualization of macrophage pyroptosis and interleukin-1β release in a viral hemorrhagic infection in zebrafish larvae. J Virol 88:12026–12040. https://doi.org/10.1128/JVI.02056-14
Weiler JM, Packard BD (1982) Methylprednisolone inhibits the alternative and amplification pathways of complement. Infect Immun 38:122–126
White TA, Perkins SE (2012) The ecoimmunology of invasive species. Funct Ecol 26:1313–1323. https://doi.org/10.2307/23326826
Yao M, Hu F, Denver RJ (2008) Distribution and corticosteroid regulation of glucocorticoid receptor in the brain of Xenopus laevis. J Comp Neurol 508:967–982. https://doi.org/10.1002/cne.21716
Zou J, Bird S, Minter R, Horton J, Cunningham C, Secombes CJ (2000) Molecular cloning of the gene for interleukin-1β from Xenopus laevis and analysis of expression in vivo and in vitro. Immunogenetics 51:332–338
Acknowledgements
We thank Tonia Schwartz, Elizabeth Schwartz, and Jeff Goessling for their feedback. We would like to acknowledge the CMB Peaks of Excellence fellowship offered by Auburn University for financial support, as well as the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) through the BEPE scholarship (2017/04802-5) for VRA (https://www.fapesp.br).
Author information
Authors and Affiliations
Contributions
Ideas and methodology were conceived by SG, VA, AA, and MTM; data was collected by SG, VA, and KS Data analysis was performed by SG, and SG led the writing of the manuscript. All authors contributed critically to the drafts and gave final approval for publication.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
Cane toads were collected under FFWC permit EXOT-17-34. All procedures performed in studies involving animals were in accordance with the ethical standards of Auburn University IACUC (PRN 2017-3106).
Additional information
Communicated by H.V. Carey.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Supplemental Fig.1
Standard metabolic rate (SMR) increase (%) of Florida cane toads (Rhinella marina) injected into the dorsal lymph sac with either LPS (20 µg g-1 body mass (strain 0111:B4)) (triangles) or saline (squares) from (A) a northern edge (New Port Richey) (n = 9,6 (LPS and saline)) and (B) a southern core (Miami) population (n = 11,5 (LPS and saline)) compared to baseline metabolic rates over a 20h period. All metabolic data were collected from toads responding to the treatments at 32oC. Dashed lines indicate no change in SMR compared to baseline trials. The graphs show mean values for each time point and error bars represent +1 SE (DOCX 93 kb)
Rights and permissions
About this article
Cite this article
Gardner, S.T., Assis, V.R., Smith, K.M. et al. Innate immunity of Florida cane toads: how dispersal has affected physiological responses to LPS. J Comp Physiol B 190, 317–327 (2020). https://doi.org/10.1007/s00360-020-01272-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00360-020-01272-7