Elucidating stygofaunal trophic web interactions via isotopic ecology

Autoři: Mattia Saccò aff001;  Alison J. Blyth aff001;  William F. Humphreys aff002;  Alison Kuhl aff004;  Debashish Mazumder aff005;  Colin Smith aff006;  Kliti Grice aff001
Působiště autorů: WA-Organic Isotope Geochemistry Centre, The Institute for Geoscience Research, School of Earth and Planetary Sciences, Curtin University, Bentley, WA (Australia) aff001;  Collections and Research Centre, Western Australian Museum, Welshpool, 6986, WA (Australia) aff002;  School of Biological Sciences, University of Western Australia, Crawley, Western Australia (Australia) aff003;  Organic Geochemistry Unit, Bristol Biogeochemistry Research Centre, School of Chemistry, University of Bristol, Bristol, United Kingdom aff004;  Australian Nuclear Science and Technology Organisation (ANSTO), Locked Bag 2001, Kirrawee DC, NSW (Australia) aff005;  Department of Archaeology and History, La Trobe University, Bundoora, VIC (Australia) aff006
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0223982


Subterranean ecosystems host highly adapted aquatic invertebrate biota which play a key role in sustaining groundwater ecological functioning and hydrological dynamics. However, functional biodiversity studies in groundwater environments, the main source of unfrozen freshwater on Earth, are scarce, probably due to the cryptic nature of the systems. To address this, we investigate groundwater trophic ecology via stable isotope analysis, employing δ13C and δ15N in bulk tissues, and amino acids. Specimens were collected from a shallow calcrete aquifer in the arid Yilgarn region of Western Australia: a well-known hot-spot for stygofaunal biodiversity. Sampling campaigns were carried out during dry (low rainfall: LR) and the wet (high rainfall: HR) periods. δ13C values indicate that most of the stygofauna shifted towards more 13C-depleted carbon sources under HR, suggesting a preference for fresher organic matter. Conversion of δ15N values in glutamic acid and phenylalanine to a trophic index showed broadly stable trophic levels with organisms clustering as low-level secondary consumers. However, mixing models indicate that HR conditions trigger changes in dietary preferences, with increasing predation of amphipods by beetle larvae. Overall, stygofauna showed a tendency towards opportunistic and omnivorous habits—typical of an ecologically tolerant community—shaped by bottom-up controls linked with changes in carbon flows. This study provides baseline biochemical and ecological data for stygofaunal trophic interactions in calcretes. Further studies on the carbon inputs and taxa-specific physiology will help refine the interpretation of the energy flows shaping biodiversity in groundwaters. This will aid understanding of groundwater ecosystem functioning and allow modelling of the impact of future climate change factors such as aridification.

Klíčová slova:

Amino acid analysis – Beetles – Copepods – Food web structure – Larvae – Predation – Stable isotopes – Trophic interactions


1. Wallace JB, Webster JR. The role of macroinvertebrates in stream ecosystem function. Annual review of entomology. 1996; 41(1):115–139.

2. Cummins K, Klug J. Feeding ecology of stream invertebrate. Annual Review Ecology and Systemic; 1979.

3. Brodersen KP, Dall PC, Lindegaard C. The fauna in the upper stony littoral of Danish lakes: macroinvertebrates as trophic indicators. Freshwater Biology. 1998; 39(3):577–592.

4. Tomanova S, Goitia E, Helešic J. Trophic levels and functional feeding groups of macroinvertebrates in neotropical streams. Hydrobiologia. 2006; 556(1):251–264.

5. Lecerf A, Usseglio-Polatera P, Charcosset JY, Lambrigot D, Bracht B, Chauvet E. Assessment of functional integrity of eutrophic streams using litter breakdown and benthic macroinvertebrates. Archiv für Hydrobiologie. 2006; 165(1):105–126.

6. Dole-Olivier MJ, Malard F, Martin D, Lefébure T, Gibert J. (2009). Relationships between environmental variables and groundwater biodiversity at the regional scale. Freshwater Biology. 2009; 54(4):797–813.

7. Korbel K, Chariton A, Stephenson S, Greenfield P, Hose GC. Wells provide a distorted view of life in the aquifer: implications for sampling, monitoring and assessment of groundwater ecosystems. Scientific reports. 2017; 7:40702. doi: 10.1038/srep40702 28102290

8. Tomlinson M, Boulton AJ, Hancock PJ, Cook PG. Deliberate omission or unfortunate oversight: Should stygofaunal surveys be included in routine groundwater monitoring programs?. Hydrogeology Journal. 2007; 15(7):1317–1320.

9. Gibert J, Culver DC, Dole‐Olivier MJ, Malard F, Christman MC, Deharveng L. Assessing and conserving groundwater biodiversity: synthesis and perspectives. Freshwater Biology. 2009; 54:930–941.

10. Strayer DL. Limits to biological distributions in groundwater. Groundwater ecology. 1994; 1:287.

11. Gibert J, Deharveng L. Subterranean Ecosystems: A Truncated Functional Biodiversity. BioScience. 2002; 52(6):473–481.

12. Hancock PJ, Boulton AJ, Humphreys WF. Aquifers and hyporheic zones: towards an ecological understanding of groundwater. Hydrogeology Journal. 2005; 13(1):98–111.

13. Humphreys WF. Hydrogeology and groundwater ecology: Does each inform the other? Hydrogeology Journal. 2009; 17(1):5–21.

14. Stoch F. Diversity in groundwaters, or: why are there so many. Mémoires de Biospéologie. 1995; 22:139–160.

15. Hartland A, Fenwick GD, Bury SJ. Tracing sewage-derived organic matter into a shallow groundwater food web using stable isotope and fluorescence signatures. Marine and Freshwater Research. 2011; 62(2):119–129.

16. Brankovits D, Pohlman JW, Niemann H, Leigh MB, Leewis MC, Becker K, et al. Methane-and dissolved organic carbon-fueled microbial loop supports a tropical subterranean estuary ecosystem. Nature communications. 2017; 8:1835. doi: 10.1038/s41467-017-01776-x 29180666

17. Hose GC, Stumpp C. Architects of the underworld: bioturbation by groundwater invertebrates influences aquifer hydraulic properties. Aquatic sciences. 2019; 81(1):20.

18. Mermillod-Blondin F, Rosenberg R. Ecosystem engineering: the impact of bioturbation on biogeochemical processes in marine and freshwater benthic habitats. Aquatic sciences. 2006; 68:434–442.

19. Murray BR, Hose GC, Eamus D, Licari D. Valuation of groundwater-dependent ecosystems: a functional methodology incorporating ecosystem services. Australian Journal of Botany. 2006; 54:221–229.

20. Boulton AJ, Fenwick GD, Hancock PJ, Harvey MS. Biodiversity, functional roles and ecosystem services of groundwater invertebrates. Invertebrate Systematics. 2008; 22:103–116.

21. Francois CM, Mermillod‐Blondin F, Malard F, Fourel F, Lécuyer C, Douady, et al. Trophic ecology of groundwater species reveals specialization in a low‐productivity environment. Functional ecology. 2016; 30(2):262–273.

22. Saccò M, Blyth A, Bateman PW, Hua Q, Mazumder D, White N, et al. New light in the dark-a proposed multidisciplinary framework for studying functional ecology of groundwater fauna. Science of the Total Environment. 2019a; 662:963–977. doi: 10.1016/j.scitotenv.2019.01.296 30795483

23. Herman PM, Middelburg JJ, Widdows J, Lucas CH, Heip CH. Stable isotopes as trophic tracers: combining field sampling and manipulative labelling of food resources for macrobenthos. Marine Ecology Progress Series. 2000; 204:79–92.

24. Boecklen WJ, Yarnes CT, Cook BA, James AC. On the use of stable isotopes in trophic ecology. Annual review of ecology, evolution, and systematics. 2011; 42:411–440.

25. Hutchins BT, Engel AS, Nowlin WH, Schwartz BF. Chemolithoautotrophy supports macroinvertebrate food webs and affects diversity and stability in groundwater communities. Ecology. 2016; 97(6):1530–1542. doi: 10.1890/15-1129.1 27459783

26. Reiss J, Perkins DM, Fussmann KE, Krause S, Canhoto C, Romeijn P, Robertson AL. Groundwater flooding: Ecosystem structure following an extreme recharge event. Science of The Total Environment. 2019; 652:1252–1260. doi: 10.1016/j.scitotenv.2018.10.216 30586811

27. Steffan SA, Chikaraishi Y, Horton DR, Ohkouchi N, Singleton ME, Miliczky E, et al. Trophic hierarchies illuminated via amino acid isotopic analysis. PLoS One. 2013; 8:e76152. doi: 10.1371/journal.pone.0076152 24086703

28. Chikaraishi Y, Steffan SA, Ogawa NO, Ishikawa NF, Sasaki Y, Tsuchiya M, et al. High‐resolution food webs based on nitrogen isotopic composition of amino acids. Ecology and evolution. 2014; 4(12):2423–2449. doi: 10.1002/ece3.1103 25360278

29. Du J, Cheung WW, Zheng X, Chen B, Liao J, Hu W. Comparing trophic structure of a subtropical bay as estimated from mass-balance food web model and stable isotope analysis. Ecological Modelling. 2015: 312: 175–181.

30. McMahon KW, Fogel ML, Elsdon TS, Thorrold SR. Carbon isotope fractionation of amino acids in fish muscle reflects biosynthesis and isotopic routing from dietary protein. Journal of Animal Ecology. 2010; 79(5):1132–1141. doi: 10.1111/j.1365-2656.2010.01722.x 20629794

31. Chikaraishi Y, Ogawa NO, Ohkouchi N. Further evaluation of the trophic level estimation based on nitrogen isotopic composition of amino acids. Earth, life, and isotopes. 2010; 37–51.

32. Choy CA, Davison PC, Drazen JC, Flynn A, Gier EJ, Hoffman JC et al. Global trophic position comparison of two dominant mesopelagic fish families (Myctophidae, Stomiidae) using amino acid nitrogen isotopic analyses. PLoS One. 2012; 7(11):e50133. doi: 10.1371/journal.pone.0050133 23209656

33. Chikaraishi Y, Kashiyama Y, Ogawa NO, Kitazato H, Ohkouchi N. Metabolic control of nitrogen isotope composition of amino acids in macroalgae and gastropods: implications for aquatic food web studies. Marine Ecology Progress Series. 2007; 342:85–90.

34. Potapov AM, Tiunov AV, Scheu S, Larsen T, Pollierer MM. Combining bulk and amino acid stable isotope analyses to quantify trophic level and basal resources of detritivores: a case study on earthworms. Oecologia. 2019; 189(2):447–460. doi: 10.1007/s00442-018-04335-3 30659383

35. Whiteman JP, Elliott Smith EA, Besser AC, Newsome SD. A Guide to Using Compound-Specific Stable Isotope Analysis to Study the Fates of Molecules in Organisms and Ecosystems. Diversity. 2019; 11(1):8.

36. Allford A, Cooper SJ, Humphreys WF, Austin AD. Diversity and distribution of groundwater fauna in a calcrete aquifer: does sampling method influence the story? Invertebrate Systematics. 2008; 22(2):127–138.

37. Hyde J, Cooper SJ, Humphreys WF, Austin AD, Munguia P. Diversity patterns of subterranean invertebrate fauna in calcretes of the Yilgarn Region, Western Australia. Marine and Freshwater Research. 2018; 69(1):114–121.

38. Saccò M, Blyth A, Humphreys WF, Karasiewicz S, Meredith K, Laini A, et al. Stygofaunal community trends along varied rainfall conditions: deciphering ecological niche dynamics of a shallow calcrete in Western Australia. Ecohydrology. 2019b; in press.

39. Gray DJ, Noble RR, Reid N, Sutton GJ, Pirlo MC. Regional scale hydrogeochemical mapping of the northern Yilgarn Craton, Western Australia: a new technology for exploration in arid Australia. Geochemistry: Exploration, Environment, Analysis. 2016; 16 (1):100–115.

40. Bradford TM, Humphreys WF, Austin AD, Cooper SJ. Identification of trophic niches of subterranean diving beetles in a calcrete aquifer by DNA and stable isotope analyses. Marine and Freshwater Research. 2014; 65(2):95–104.

41. Mazumder D, Saintilan N, Wen L, Kobayashi T, Rogers K. Productivity influences trophic structure in a temporally forced aquatic ecosystem. Freshwater Biology. 2017; 62(9):1528–1538.

42. Dogramaci S, Skrzypek G. Unravelling sources of solutes in groundwater of an ancient landscape in NW Australia using stable Sr, H and O isotopes. Chemical Geology. 2015; 393:67–78.

43. Mora A, Arriaza BT, Standen VG, Valdiosera C, Salim A, Smith C. High resolution palaeodietary reconstruction: amino acid d13C analysis of keratin from single hairs of mummified human individuals. Quaternary International. 2017; 436:96e113.

44. Smith CI, Fuller BT, Choy K, Richards MP. A three-phase liquid chromatographic method for d13C analysis of amino acids from biological protein hydrolysates using liquid chromatography isotope ratio mass spectrometry. Anal. Biochem. 2009; 390:165e172.

45. Mora A, Pacheco A, Roberts C, Smith C. Pica 8: Refining dietary reconstruction through amino acid δ13C analysis of tendon collagen and hair keratin. Journal of Archaeological Science. 2018; 93:94–109.

46. Styring AK, Kuhl A, Knowles TD, Fraser RA, Bogaard A, Evershed RP. Practical considerations in the determination of compound‐specific amino acid δ15N values in animal and plant tissues by gas chromatography‐combustion‐isotope ratio mass spectrometry, following derivatisation to their N‐acetylisopropyl esters. Rapid Communications in Mass Spectrometry. 2012; 26(19):2328–2334. doi: 10.1002/rcm.6322 22956325

47. Boudko DY. Molecular basis of essential amino acid transport from studies of insect nutrient amino acid transporters of the SLC6 family (NAT-SLC6). Journal of insect physiology. 2012; 58(4):433–449. doi: 10.1016/j.jinsphys.2011.12.018 22230793

48. Fantle MS, Dittel AI, Schwalm SM, Epifanio CE, Fogel ML. A food web analysis of the juvenile blue crab, Callinectes sapidus, using stable isotopes in whole animals and individual amino acids. Oecologia. 1999; 120:416–426. doi: 10.1007/s004420050874 28308018

49. Gómez C, Larsen T, Popp B, Hobson KA, Cadena CD. Assessing seasonal changes in animal diets with stable-isotope analysis of amino acids: a migratory boreal songbird switches diet over its annual cycle. Oecologia. 2018; 187(1):1–13. doi: 10.1007/s00442-018-4113-7 29564539

50. Newsome SD, Fogel ML, Kelly L, del Rio CM. Contributions of direct incorporation from diet and microbial amino acids to protein synthesis in Nile tilapia. Functional Ecology. 2011; 25(5):1051–1062.

51. Steffan S.A., Chikaraishi Y., Horton D.R., Ohkouchi N., Singleton M.E., Miliczky et al. Trophic hierarchies illuminated via amino acid isotopic analysis. PLoS One 8. 2013; e76152.

52. Post DM. Using stable isotopes to estimate trophic position: models, methods, and assumptions. Ecology. 2002; 83(3):703–718.

53. Galassi DM, Huys R, Reid JW. Diversity, ecology and evolution of groundwater copepods. Freshwater Biology. 2009; 54(4):691–708.

54. Foulquier A, Malard F, Mermillod-Blondin F, Montuelle B, Dolédec S, Volat B, et al. Surface water linkages regulate trophic interactions in a groundwater food web. Ecosystems. 2011; 14(8):1339–1353.

55. Newsome SD, Wolf N, Peters J, Fogel ML. Amino acid δ13C analysis shows flexibility in the routing of dietary protein and lipids to the tissue of an omnivore. Integrative and Comparative Biology. 2014; 54:890–902 doi: 10.1093/icb/icu106 25104856

56. Jim S, Jones V, Ambrose SH, Evershed RP. Quantifying dietary macronutrient sources of carbon for bone collagen biosynthesis using natural abundance stable carbon isotope analysis. British Journal of Nutrition. 2006; 95:1055–1062. doi: 10.1079/bjn20051685 16768826

57. Pohlman J. W. The biogeochemistry of anchialine caves: progress and possibilities. Hydrobiologia. 2011; 677(1):33–51.

58. Datry T, Malard F, Gibert J. Response of invertebrate assemblages to increased groundwater recharge rates in a phreatic aquifer. Journal of the North American Benthological Society. 2005; 24:461–477.

59. Danielopol DL. Groundwater fauna associated with riverine aquifers. Journal of the North American Benthological Society. 1989; 8(1):18–35.

60. Mekhanikova IV. Morphology of mandible and lateralia in six endemic amphipods (Amphipoda, Gammaridea) from Lake Baikal, in relation to feeding. Crustaceana. 2010; 83:865–887.

61. Žutinić P, Petrić I, Gottstein S, Udovič MG, Borojević KK, Kamberović, et al. Microbial mats as shelter microhabitat for amphipods in an intermittent karstic spring. Knowledge & Management of Aquatic Ecosystems. 2018; 419:7.

62. Saint-Marie B. Morphological adaptations for carrion feeding in four species of littoral or circalittoral lysianassid amphipods. Canadian Journal of Zoology. 1984; 62:1668–1674.

63. Friberg N, Jacobsen D. Feeding plasticity of two detritivore-shredders. Freshwater Biology. 1994; 32(1):133–142.

64. Hutchins BT, Schwartz BF, Nowlin WH. Morphological and trophic specialization in a subterranean amphipod assemblage. Freshwater biology. 2014; 59(12):2447–2461.

65. Jasinska EJ, Knott B, McComb AJ. Root mats in ground water: a fauna-rich cave habitat. Journal of the North American Benthological Society. 1996; 15(4):508–519.

66. Navel S, Simon L, Lécuyer C, Fourel F, Mermillod‐Blondin F. The shredding activity of gammarids facilitates the processing of organic matter by the subterranean amphipod Niphargus rhenorhodanensis. Freshwater Biology. 2011; 56(3):481–490.

67. Simon KS, Benfield EF, Macko SA. Food web structure and the role of epilithic biofilms in cave streams. Ecology. 2003; 84(9):2395–2406.

68. Sinha A, Aubry MP, Stott L, Thiry M, Bergreen WA. Chemostratigraphy of the “lower” Sparnacian deposits (Argiles plastiques bariolées) of the Paris Basin. Israel Journal of Earth Science. 1996; 44:223–237.

69. Thiry M, Aubry MP, Dupuis C, Sinha A, Stott LD, Berggren WA. The Sparnacian deposits of the Paris Basin: d13C isotope stratigraphy. Stratigraphy. 2006; 3(2):119–138.

70. Galy V, Bouchez J, France‐Lanord C. Determination of total organic carbon content and δ13C in carbonate‐rich detrital sediments. Geostandards and Geoanalytical research. 2007; 31(3):199–207.

71. Cartwright I, Hannam K, Weaver TR. Constraining flow paths of saline groundwater at basin margins using hydrochemistry and environmental isotopes: Lake Cooper, Murray Basin, Australia. Australian Journal of Earth Sciences. 2007; 54(8):1103–1122.

72. Bryan E, Meredith KT, Baker A, Andersen MS, Post VE. (2017). Carbon dynamics in a Late Quaternary-age coastal limestone aquifer system undergoing saltwater intrusion. Science of the Total Environment. 2017; 607:771–785. doi: 10.1016/j.scitotenv.2017.06.094 28711007

73. Lipar M., Webb JA, Cupper ML, Wang N. Aeolianite, calcrete/microbialite and karst in southwestern Australia as indicators of Middle to Late Quaternary palaeoclimates. Palaeogeography, palaeoclimatology, palaeoecology. 2017; 470:11–29.

74. Portillo MC, Porca E, Cuezva S, Canaveras JC, Sanchez-Moral S, Gonzalez JM. Is the availability of different nutrients a critical factor for the impact of bacteria on subterraneous carbon budgets?. Naturwissenschaften. 2009; 96(9):1035–1042. doi: 10.1007/s00114-009-0562-5 19488732

75. Chapelle FH. The significance of microbial processes in hydrogeology and geochemistry. Hydrogeology Journal. 2000; 8(1):41–46.

76. Flynn TM, Sanford RA, Ryu H, Bethke CM, Levine AD, Ashbolt NJet al. (2013). Functional microbial diversity explains groundwater chemistry in a pristine aquifer. BMC microbiology. 2013; 13(1):146.

77. Kutvonen H, Rajala P, Carpén L, Bomberg M. Nitrate and ammonia as nitrogen sources for deep subsurface microorganisms. Frontiers in microbiology. 2015; 6:1079. doi: 10.3389/fmicb.2015.01079 26528251

78. Thorp JH, Bowes RE. Carbon sources in riverine food webs: new evidence from amino acid isotope techniques. Ecosystems. 2017; 20(5):1029–1041.

79. Edmundus WM, Smedley PL. Groundwater geochemistry and health: an overview. Geological Society, London, Special Publications. 1996; 113(1):91–105.

80. Jones KK, Cooper SJ, Seymour RS. Cutaneous respiration by diving beetles from underground aquifers of Western Australia (Coleoptera: Dytiscidae). Journal of Experimental Biology. 2019; 222(7):jeb196659.

81. Lundkvist E, Landin J, Jackson M, Svensson C. Diving beetles (Dytiscidae) as predators of mosquito larvae (Culicidae) in field experiments and in laboratory tests of prey preference. Bulletin of entomological research. 2003; 93(3):219–226. doi: 10.1079/BER2003237 12762863

82. Leijs R, Van Nes EH, Watts CH, Cooper SJ, Humphreys WF, Hogendoorn K. Evolution of blind beetles in isolated aquifers: a test of alternative modes of speciation Plos One. 2012; 7(3):e34260. doi: 10.1371/journal.pone.0034260 22479581

83. Alarie Y, Michat MC, Watts CH. Larval morphology of Paroster Sharp, 1882 (Coleoptera: Dytiscidae: Hydroporinae): reinforcement of the hypothesis of monophyletic origin and discussion of phenotypic accommodation to a hypogaeic environment. Zootaxa. 2009; 2274:1–44.

84. Inoda T. Predaceous diving beetle, Dytiscus sharpi sharpi (Coleoptera: Dytiscidae) larvae avoid cannibalism by recognizing prey. Zoological science. 2012; 29(9):547–553. doi: 10.2108/zsj.29.547 22943777

85. Shen Y, Chapelle FH, Strom EW, Benner R. Origins and bioavailability of dissolved organic matter in groundwater. Biogeochemistry. 2015; 122(1):61–78.

86. Simon KS, Pipan T, Culver DC. A conceptual model of the flow and distribution of organic carbon in caves. Journal of Cave and Karst Studies. 2007; 69(2):279–284.

87. Elton C. Animal ecology. Sidwick and Jackson, London, England; 1927.

88. Hunter MD, Price PW. Playing chutes and ladders: heterogeneity and the relative roles of bottom‐up and top‐down forces in natural communities. Ecology. 1992; 73(3):724–732.

89. Walker M, Jones TH. Relative roles of top‐down and bottom‐up forces in terrestrial tritrophic plant–insect herbivore–natural enemy systems. Oikos. 2001; 93(2):177–187.

90. Lemmens P, Declerck SA, Tuytens K, Vanderstukken M, De Meester L. Bottom-up effects on biomass versus top-down effects on identity: a multiple-lake fish community manipulation experiment. Ecosystems. 2018; 21(1):166–177.

91. Foulquier A, Simon L, Gilbert F, Fourel F, Malard F, Mermillod‐Blondin F. Relative influences of DOC flux and subterranean fauna on microbial abundance and activity in aquifer sediments: new insights from 13C‐tracer experiments. Freshwater biology. 2010; 55(7):1560–1576.

92. Venarsky MP, Benstead JP, Huryn AD. Effects of organic matter and season on leaf litter colonisation and breakdown in cave streams. Freshwater Biology. 2012; 57(4):773–786.

93. Power ME. Top‐down and bottom‐up forces in food webs: do plants have primacy. Ecology. 1992; 73(3):733–746.

94. Frau D, Devercelli M, de Paggi SJ, Scarabotti P, Mayora G, Battauz Y, et al. Can top-down and bottom-up forces explain phytoplankton structure in a subtropical and shallow groundwater-connected lake?. Marine and Freshwater Research. 2015; 66(12):1106–1115.

95. Bradford T. M. Modes of speciation in subterranean diving beetles from a single calcrete aquifer in Central Western Australia (Doctoral dissertation); 2010.

96. Hyde J. C. A. Investigating the internal and external ecology of six subterranean diving beetle species from the Yilgarn region of Central Australia (Doctoral dissertation); 2010.

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