We investigated amphibious behaviour, hydrogen sulphide (H2S) tolerance, and the mechanism of H2S toxicity in the amphibious mangrove rivulus (Kryptolebias marmoratus). We found that fish emersed (left water) in response to acutely elevated [H2S] (~ 130–200 µmol l−1). The emersion response to H2S may be influenced by prior acclimation history due to acclimation-induced alterations in gill morphology and/or the density and size of neuroepithelial cells (NECs) on the gills and skin. Thus, we acclimated fish to water (control), H2S-rich water, or air and tested the hypotheses that acclimation history influences H2S sensitivity due to acclimation-induced changes in (i) gill surface area and/or (ii) NEC density and/or size. Air-acclimated fish emersed at significantly lower [H2S] relative to fish acclimated to control or H2S-rich water, but exhibited no change in gill surface area or in NEC density or size in the gills or skin. Despite possessing exceptional H2S tolerance, all fish lost equilibrium when unable to emerse from environments containing extremely elevated [H2S] (2272 ± 46 µmol l−1). Consequently, we tested the hypothesis that impaired blood oxygen transport (i.e., sulphemoglobin formation) causes H2S toxicity in amphibious fishes. In vitro exposure of red blood cells to physiologically relevant [H2S] did not cause a substantial increase in sulphemoglobin formation. We found evidence, however, for an alternative hypothesis that H2S toxicity is caused by impaired oxidative phosphorylation (i.e., cytochrome c oxidase inhibition). Collectively, our results show that amphibious behaviour is critical for the survival of K. marmoratus in H2S-rich environments as fish experience impaired oxidative phosphorylation when unable to emerse.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Abel DC, Koenig CC, Davis WP (1987) Emersion in the mangrove forest fish Rivulus marmoratus: a unique response to hydrogen sulfide. Environ Biol Fish 18:67–72
Bagarinao T (1992) Sulfide as an environmental factor and toxicant: tolerance and adaptations in aquatic organisms. Aquat Toxicol 24:21–62
Bagarinao T, Vetter RD (1989) Sulfide tolerance and detoxification in shallow-water marine fishes. Mar Biol 103:291–302
Bagarinao T, Vetter RD (1992) Sulfide-hemoglobin interactions in the sulfide-tolerant salt marsh resident, the California killifish Fundulus parvipinnis. J Comp Physiol B 162:614–624
Bianchini K, Wright PA (2013) Hypoxia delays hematopoiesis: retention of embryonic hemoglobin and erythrocytes in larval rainbow trout, Oncorhynchus mykiss, during chronic hypoxia exposure. J Exp Biol 216:4415–4425
Blanchard TS, Whitehead A, Dong YW, Wright PA (2019) Phenotypic flexibility in respiratory traits is associated with improved aerial respiration in an amphibious fish out of water. J Exp Biol 222(2):jeb186486
Borowiec BG, Darcy KL, Gillette DM, Scott GR (2015) Distinct physiological strategies are used to cope with constant hypoxia and intermittent hypoxia in killifish (Fundulus heteroclitus). J Exp Biol 218:1198–1211
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Brauner CJ, Ballantyne CL, Randall DJ, Val AL (1995) Air breathing in the armoured catfish (Hoplosternum littorale) as an adaptation to hypoxic, acidic, and hydrogen sulphide rich waters. Can J Zool 73:739–744
Broderius SJ, Smith LLJ (1977) Direct determination and calculation of aqueous hydrogen sulfide. Anal Chem 49:424–428
Cline JD (1969) Spectrophotometric determination of hydrogen sulfide in natural waters. Limnol Oceanogr 14:454–458
Davenport J, Woolmington AD (1981) Behavioural responses of some rocky shore fish exposed to adverse environmental conditions. Mar Freshw Behav Phy 8:1–12
Ebeling AW, Bernal P, Zuleta A (1970) Emersion of the amphibious chilean clingfish, Sicyases sanguineus. Biol Bull 139:115–137
Frick NT, Wright PA (2002a) Nitrogen metabolism and excretion in the mangrove killifish Rivulus marmoratus I. The influence of environmental salinity and external ammonia. J Exp Biol 205:79–89
Frick NT, Wright PA (2002b) Nitrogen metabolism and excretion in the mangrove killifish Rivulus marmoratus II. Significant ammonia volatilization in a teleost during air-exposure. J Exp Biol 205:91–100
Geiger SP, Torres JJ, Crabtree RE (2000) Air breathing and gill ventilation in juvenile tarpon, Megalops atlanticus: responses to changes in dissolved oxygen, temperature, hydrogen sulfide, and pH. Environ Biol Fish 59:181–190
Gibson DJ, Sylvester EVA, Turko AJ, Tattersall GJ, Wright PA (2015) Out of the frying pan into the air—emersion behaviour and evaporative heat loss in an amphibious mangrove fish (Kryptolebias marmoratus). Biol Lett. https://doi.org/10.1098/rsbl.2015.0689
Ip YK, Kuah SSL, Chew SF (2004) Strategies adopted by the mudskipper Boleophthalmus boddaerti to survive sufide exposure in normoxia or hypoxia. Physiol Biochem Zool 77:824–837
Kelley JL, Arias-Rodriguez L, Martin DP, Yee MC, Custamante CD, Tobler M (2016) Mechanisms underlying adaptation to life in hydrogen sulfide-rich environments. Mol Biol Evol 33:1419–1434
Lau GY, Mandic M, Richards JG (2017) Evolution of cytochrome c oxidase in hypoxia tolerant sculpins (Cottidae, Actinopterygii). Mol Biol Evol 34:2153–2162
Milsom WK, Burleson ML (2007) Peripheral arterial chemoreceptors and the evolution of the carotid body. Resp Physiol Neurobi 157:4–11
Miwa S, Iunui Y (1991) Thyroid hormone stimulates the shift of erythrocyte populations during metamorphosis of the flounder. J Exp Zool 259:222–228
Olson KR, Healy MJ, Qin Z, Skovgaard N, Vulesevic B, Duff DW, Whitfield NL, Yang G, Wang R, Perry SF (2008) Hydrogen sufide as an oxygen sensor in trout gill chemoreceptors. Am J Physiol Reg I 295:669–680
Ong KJ, Stevens ED, Wright PA (2007) Gill morphology of the mangrove killifish (Kryptolebias marmoratus) is plastic and changes in response to terrestrial air exposure. J Exp Biol 210:1109–1115
Patrick ML, Pärt P, Marshall WS, Wood CM (1997) Characterization of ion and acid-base transport in the fresh water adapted mummichog (Fundulus heteroclitus). J Exp Zool 279:208–219
Pfenninger M, Lerp H, Tobler M, Passow C, Kelley JL, Funke E, Greshake B, Erkoc UK, Berberich T, Plath M (2014) Parallel evolution of cox genes in H2S-tolerant fish as key adaptation to a toxic environment. Nat Commun 5:3873–3873
Pietri R, Román-Morales E, López-Garriga J (2011) Hydrogen sulfide and hemeproteins: knowledge and mysteries. Antioxid Redox Sign 15:393–404
Plath M, Tobler M, Riesch R, García de León FJ, Giere O, Schlupp I (2007) Survival in an extreme habitat: the role of behaviour and energy limitation. Naturwissenschaften 94:991–996
Porteus CS, SAbdallah SJ, Pollack J, Kumai Y, Kwong RWM, Yew HM, Milsom WK, Perry SF (2014) The role of hydrogen sulphide in the control of breathing in hypoxic zebrafish (Danio rerio). J Physiol 592:3075–3088
Regan KS, Jonz MG, Wright PA (2011) Neuroepithelial cells and the hypoxia emersion response in amphibious fish Kryptolebias marmoratus. J Exp Biol 214:2560–2568
Regan MD, Gill IS, Richards JG (2017) Metabolic depression and the evolution of hypoxia tolerance in threespine stickleback, Gasterosteus aculeatus. Biol Lett. https://doi.org/10.1098/rsbl.2017.0392
Riesch R, Plath M, Schlupp I (2010) Toxic hydrogen sulfide and dark caves: life-history adaptation in a livebearing fish (Poecilia mexicana, Poeciliidae). Ecology 91:1494–1505
Riesch R, Schlupp I, Langerhands RB, Plath M (2011) Shared and unique patterns of embryo development in extremophile Poeciliids. PLoS One. https://doi.org/10.1371/journal.pone.0027377
Riesch R, Tobler M, Plath M (2015) Hydrogen sulfide-toxic habitats. In: Riesch R, Tobler M, Plath M (eds) Extremophile fishes. Springer International Publishing, Switzerland, pp 137–159
Ritchie SA, Johnson ES (1986) Modified Gee’s improved wire minnow trap: an excellent surveillance tool for mosquito larvivores. J Florida Anti-Mosq 57:22–24
Robertson CE, Turko AJ, Jonz MG, Wright PA (2015) Hypercapnia and low pH induce neuroepithelial cell proliferation and emersion behaviour in the amphibious fish Kryptolebias marmoratus. J Exp Biol 218:2987–2990
Rossi GS, Tunnah L, Martin KE, Turko AJ, Taylor DS, Currie S, Wright PA (2019) Mangrove fishes rely on emersion behaviour and physiological tolerance to persist in sulfidic environments. Physiol Biochem Zool (in press)
Saltys HA, Jonz MG, Nurse CA (2006) Comparative study of gill neuroepithelial cells and their innervation in teleosts and Xenopus tadpoles. Cell Tissue Res 323:1–10
Tatarenkov A, Ring BC, Elder JF, Bechler DL, Avise JC (2010) Genetic composition of laboratory stocks of the self-fertilizing fish Kryptolebias marmoratus: a valuable resource for experimental research. PLoS One. https://doi.org/10.1371/journal.pone.0012863
Taylor DS (1990) Adaptive specializations of the cyprinodont fish Rivulus marmoratus. Fla Sci 53:239–248
Taylor DS (2012) Twenty-four years in the mud: what have we learned about the natural history and ecology of the mangrove rivulus, Kryptolebias marmoratus? Integr Comp Biol 52:724–736
Tobler M, DeWitt TJ, Schlupp I, García de León FJ, Herrmann R, Fuelner PGD, Tiedemann R, Plath M (2008) Toxic hydrogen sulfide and dark caves: phenotypic and genetic divergence across two abiotic environmental gradients in Poecilia mexicana. Evolution 62:2643–2659
Tobler M, Palacios M, Chapman LJ, Mitrofanov I, Bierbach D, Plath M, Arias-Rodriguez L, García de León F, Mateos M (2011) Evolution in extreme environments: replicated phenotypic differentiation in livebearig fish inhabiting sulfidic springs. Evolution 65:2213–2228
Tobler M, Kelley JL, Plath M, Riesch R (2018) Extreme environments and the origins of biodiversity: adaptation and speciation in sulphide spring fishes. Mol Ecol 27:843–859
Turko AJ, Wright PA (2015) Evolution, ecology and physiology of amphibious killifishes (Cyprinodontiformes). J Fish Biol 87:815–835
Turko AJ, Early RL, Wright PA (2011) Behaviour drives morphology: voluntary emersion patterns shape gill strcutre in genetically identical mangrove rivulus. Anim Behav 82:39–47
Turko AJ, Cooper CA, Wright PA (2012) Gill remodeling during terrestrial acclimation reduces aquatic respiratory function of the amphibious fish Kryptolebias marmoratus. J Exp Biol 215:3973–3980
Turko AJ, Tatarenkov A, Currie S, Earley RL, Platek A, Taylor DS, Wright PA (2018) Emersion behaviour underlies variation in gill morphology and aquatic respiratory function in the amphibious fish Kryptolebias marmoratus. J Exp Biol 221:jeb.168039
Turko AJ, Maini P, Wright PA, Standen EM (2019) Gill remodeling during terrestrial acclimation in the amphibious fish Polypterus senegalus. J Morphol. https://doi.org/10.1002/jmor.20946
Urbina MA, Forster ME, Glover CN (2011) Leap of faith: Voluntary emersion behaviour and physiological adaptation to aerial exposure in a non-aestivating freshwater fish in response to aquatic hypoxia. Physiol Behav 103:240–247
Völkel S, Berenbrink M (2000) Sulphaemoglobin formation in fish: a comparison between the haemoglobin of the sulphide-sensitive rainbow trout (Oncorhynchus mykiss) and the sulphide-tolerant common carp (Cyprinus carpio). J Exp Biol 203:1047–1058
Wilson JM, Bunte RM, Carty AJ (2009) Evaluation of rapid cooling and tricaine methanesulfonate (MS222) as methods of euthanasia in zebrafish (Danio rerio). J Am Assoc Lab Anim 48:785–789
Zaccone G, Lauriano ER, Kuciel M, Capillo G, Pergolizzi S, Aleschi A, Ishimatsu A, Ip YW, Icardo JM (2017) Identification and distribution of neuronal nitric oxide synthase and neurochemical markers in the neuroepithelial cells of the gill and the skin in the giant mudskipper, Periophthalmodon schlosseri. Zoology 125:41–52
We thank Wen Pan for her help with confocal imaging as well as Matt Cornish, Mike Davies, Abiran Sritharan, and numerous undergraduate volunteers for assistance with animal care.
This work was supported by the National Sciences and Engineering Research Council of Canada (NSERC) Discovery grants to P.A.W (Grant number 120513).
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Communicated by H.V. Carey.
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Cochrane, P.V., Rossi, G.S., Tunnah, L. et al. Hydrogen sulphide toxicity and the importance of amphibious behaviour in a mangrove fish inhabiting sulphide-rich habitats. J Comp Physiol B 189, 223–235 (2019). https://doi.org/10.1007/s00360-019-01204-0
- Hydrogen sulphide
- Amphibious fish
- Cytochrome c oxidase