Marine Biology

, 165:118 | Cite as

Cleaner shrimp are true cleaners of injured fish

  • David B. VaughanEmail author
  • Alexandra S. Grutter
  • Hugh W. Ferguson
  • Rhondda Jones
  • Kate S. Hutson
Original paper


Reef fishes sustain injuries from various behavioural and environmental interactions. Injured fishes have been observed frequenting cleaning stations to be attended by different cleaner fishes. This symbiotic relationship between injured fishes and cleaner fishes has only been observed in the wild and has never been demonstrated empirically for cleaner shrimp. We present the first investigation of cleaning of injured fish by cleaner shrimp, and the first controlled laboratory trial investigating the functional relationship between injured client fish and cleaners. We tested whether the cleaner shrimp (Lysmata amboinensis) cleaned injured sea goldies (Pseudanthias squamipinnis) following a standardised, unilateral superficial skin lesion. We recorded the cleaning behaviour between shrimp and fish and determined that the fish regulated the cleaning and reduced the amount of cleaning time by the shrimp of the injured side immediately post-injury, corresponding with previous literary evidence of the rapid onset of re-epithelialisation of the injury to seal it in the first 24 h in injured fishes. Thereafter, injured fish showed no cleaning preference between injured and uninjured sides. Image analyses determined that the cleaner shrimp reduced the redness of the injury, representing rubor, associated with the inflammatory response in fishes. Injuries in fishes are susceptible to invasion by secondary pathogens, and the reduction of injury rubor by shrimp may suggest that cleaning by these shrimp could reduce the success of opportunistic infection. Cleaner shrimp neither aggravated existing injury, nor created additional injury, measured quantitatively. The cleaning of injured fish by cleaner shrimp thus likely involves true cleaning behaviour.



DBV was funded through an IPRS (Australian government) scholarship awarded by James Cook University, Townsville, Australia. We are grateful to Cairns Marine, Cairns, Australia, and in particular to Laura Simmons for the ongoing support for our investigations into cleaning symbiosis. We acknowledge the generous assistance of volunteers Alejandro Trujillo González, Thienthanh Trinh, Julian Wilson, and Pauline Narvaez. We thank Peter Waldie and an undisclosed reviewer for their insightful comments and suggestions that ultimately strengthened the manuscript. This paper is dedicated in memoriam to Patrick Donnelly.

Compliance with ethical standards

Statement on the welfare of animals

All applicable international, national and institutional guidelines for the care and use of animals were followed. Animal housing was inspected by the James Cook University welfare officer, and ethics approval was granted prior to commencement of this study under the James Cook University Ethics Committee Permit number A2260, conforming strictly to the national regulations set out in the National Health and Medical Research Council (2013) Australian code for the care and use of animals for scientific purposes, 8th edition, under Section 39 of the National Health and Medical Research Council Act, 1992. A superficial, standardised epithelial injury was necessary to answer the research question. Fish were given a single unilateral superficial injury by removal of scales from a predetermined area on either flank of an area no larger than 0.15 cm2. All fish were housed separately to avoid bullying. Handling of all fish briefly with an aquarium hand-held net was crucial for the transferral into and out of the experimental tanks and could not be avoided. Hand-netting was kept to the operational minimum, occurring only once per day in and out of the experimental tanks per individual. Fish were offered food daily after experimentation and fed successfully, demonstrating a rapid return to normal behaviour after brief handling. Anaesthesia was justifiably employed to facilitate rapid photography of all fish and to avoid prolonged aerial exposure. Three fish mortalities occurred during the experimentation period employing 126 fish. All fish were humanely euthanised after sampling using an anaesthetic overdose of 2-phenoxyethanol (1.5 ml/l > 10 min; Neiffer and Stamper 2009) following the approved animal end-point set out in the animal ethics approval. This article does not contain any studies with human participants performed by any of the authors.

Conflict of interest

The authors declare no conflicts of interest.

Data availability

All datasets used to generate the statistical analyses are available from

Supplementary material

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Supplementary material 1 (PDF 490 kb)
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Supplementary material 2 (PDF 135 kb)
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Supplementary material 3 (PDF 210 kb)
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Supplementary material 4 (PDF 114 kb)
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Supplementary material 5 (PDF 67 kb)


  1. Arnal C, Morand S (2001) Importance of ectoparasites and mucus in cleaning interactions in the Mediterranean cleaner wrasse Symphodus melanocercus. Mar Biol 138:777–784CrossRefGoogle Scholar
  2. Arnal C, Côté IM, Morand S (2001) Why clean and be cleaned? The importance of client ectoparasites and mucus in a marine cleaning symbiosis. Behav Ecol Sociobiol 51:1–7CrossRefGoogle Scholar
  3. Bates D, Maechler M, Bolker B, Walker S (2015) Fitting linear mixed-effects models using lme4. J Stat Soft 67:1–48CrossRefGoogle Scholar
  4. Becker JHA, Grutter AS (2004) Cleaner shrimp do clean. Coral Reefs 23:515–520Google Scholar
  5. Bertoncini AA, Machado LF, Barreiros JP, Hostim-Silva M, Verani JR (2009) Cleaning activity among Labridae in the Azores: the rainbow wrasse Coris julis and the Azorean blue wrasse Centrolabrus caeruleus. J Mar Biol Assoc UK 89:859–861CrossRefGoogle Scholar
  6. Böckelmann PK, Ochandio BS, Bechara IJ (2010) Histological study of the dynamics in epidermis regeneration of the carp tail fin (Cyprinus carpio, Linnaeus, 1758). Braz J Biol 70:217–223CrossRefPubMedGoogle Scholar
  7. Bshary R (2003) The cleaner wrasse, Labroides dimidiatus, is a key organism for reef fish diversity at Ras Mohammed national Park, Egypt. J Anim Ecol 72:169–176CrossRefGoogle Scholar
  8. Bshary R, Grutter AS (2002) Asymmetric cheating opportunities and partner control in the cleaner fish mutualism. Anim Behav 63:547–555CrossRefGoogle Scholar
  9. Bshary R, Grutter AS (2005) Punishment and partner switching cause cooperative behaviour in a cleaning mutualism. Biol Lett 1:396–399CrossRefPubMedPubMedCentralGoogle Scholar
  10. Bshary R, Oliviera RF, Oliviera TSF, Canário AVM (2007) Do cleaning organisms reduce the stress response of client reef fish? Front Zool. PubMedPubMedCentralCrossRefGoogle Scholar
  11. Bunkley-Williams L, Williams EH Jr (1998) Ability of Pederson cleaner shrimp to remove juveniles of the parasitic cymothoid isopod, Anilocra haemuli, from the host. Crustaceana 71:862–869CrossRefGoogle Scholar
  12. Castro R, Zou J, Secombes CJ, Martin SAM (2011) Cortisol modulates the induction of inflammatory gene expression in a rainbow trout macrophage cell line. Fish Shellfish Immunol 30:215–223CrossRefPubMedGoogle Scholar
  13. Chapuis L, Bshary R (2009) Strategic adjustment of service quality to client identity in the cleaner shrimp, Periclimenes longicarpus. Anim Behav 78:455–459CrossRefGoogle Scholar
  14. Cheney KL, Côté IM (2005) Mutualism or parasitism? The variable outcome of cleaning symbioses. Biol Lett 1:162–165CrossRefPubMedPubMedCentralGoogle Scholar
  15. Côté IM (2000) Evolution and ecology of cleaning symbioses in the sea. In: Gibson RN, Barnes M (eds) Oceanography and marine biology: an annual review, vol 38. Taylor and Francis, New York, pp 311–356Google Scholar
  16. Davis MW, Ottmar ML (2006) Wounding and reflex impairment may be predictors for mortality in discarded or escaped fish. Fish Res 82:1–6CrossRefGoogle Scholar
  17. Davis MW, Stephenson J, Noga EJ (2008) The effect of Tricaine on use of the fluorescein test for detecting skin and corneal ulcers in fish. J Aquat Anim Health 20:86–95CrossRefPubMedGoogle Scholar
  18. Eckes M, Dove S, Siebeck UE, Grutter AS (2015) Fish mucus versus parasitic gnathiid isopods as sources of energy and sunscreens for a cleaner fish. Coral Reefs 34:823–833CrossRefGoogle Scholar
  19. Ferreire R, Bronstein JL, Rinaldi S, Law R, Gauduchon M (2001) Cheating and the evolutionary stability of mutualisms. Proc R Soc Lond B Biol Sci 269:773–780CrossRefGoogle Scholar
  20. Fontenot DK, Neiffer DL (2004) Wound management in teleost fish: biology of the healing process, evaluation, and treatment. Vet Clin N Am Exot Anim Pract 7:57–86CrossRefGoogle Scholar
  21. Foster SA (1985) Wound healing: a possible role of cleaning stations. Copeia 4:875–880CrossRefGoogle Scholar
  22. Francini-Filho RB, Moura RL, Sazima I (2000) Cleaning by the wrasse Thalassoma norohanum, with two records of predation by its grouper client Cephalopholis fulva. J Fish Biol 56:802–809Google Scholar
  23. Gorlick DL (1980) Ingestion of host fish surface mucus by the Hawaiian cleaning wrasse, Labroides phthirophagus (Labridae), and its effect on host species preference. Copeia 4:863–868CrossRefGoogle Scholar
  24. Grutter AS (1997) Effect of the removal of cleaner fish on the abundance and species composition of reef fish. Oecologia 11:137–143CrossRefGoogle Scholar
  25. Grutter AS (2001) Parasite infection rather than tactile stimulation is the proximate cause of cleaning behaviour in reef fish. Proc R Soc Lond B Biol Sci 268:1361–1365CrossRefGoogle Scholar
  26. Grutter AS, Bshary R (2003) Cleaner wrasse prefer client mucus: support for partner control mechanisms in cleaning interactions. Proc R Soc Lond B Biol Sci 270:S242–S244CrossRefGoogle Scholar
  27. Grutter AS, Murphy JM, Choat JH (2003) Cleaner fish drives local fish diversity on coral reefs. Curr Biol 13:64–67CrossRefPubMedGoogle Scholar
  28. Grutter AS, De Brauwer M, Bshary R, Cheney KL, Cribb TH, Madin EMP, McClure EC, Meekan MG, Sun D, Warner RR, Werminghausen J, Sikkel P (2018) Parasite infestation increases on coral reefs without cleaner fish. Coral Reefs 37:15–24CrossRefGoogle Scholar
  29. Harrell FE Jr (2017) Hmisc: harrell miscellaneous. R package version 4.0-3. (with contributions from Charles Dupont and many others). Accessed 20 Sept 2017
  30. Hobson ES (1971) Cleaning symbiosis among California inshore fishes. Fish Bull 69:491–523Google Scholar
  31. Hutson KS, Brazenor AK, Vaughan DB, Trujillo-Gonzalez A (2018) Monogenean parasite cultures: current techniques and recent advances. Adv Parasitol. CrossRefPubMedGoogle Scholar
  32. Iger Y, Balm PHM, Jenner HA, Wendelaar Bonga SE (1995) Cortisol induced stress-related changes in the skin of rainbow trout (Oncorhynchus mykiss). Gen Comp Endocrinol 97:188–198CrossRefPubMedGoogle Scholar
  33. Jensen LB, Wahli T, McGurk C, Eriksen TB, Obach A, Waagbø R, Handler A, Tafalla C (2015) Effect of temperature and diet on wound healing in Atlantic salmon (Salmo salar L.). Fish Physiol Biochem 41:1527–1543CrossRefPubMedGoogle Scholar
  34. Kaneko JJ II, Yamada R, Brock JA, Nakamura RM (1988) Infection of tilapia, Oreochromis mossambicus (Trewavas), by a marine monogenean, Neobenedenia melleni (MacCallum, 1927) Yamaguti, 1963 in Kaneohe Bay, Hawaii, USA, and its treatment. J Fish Dis 11:295–300CrossRefGoogle Scholar
  35. Kolm N, Hoffman EA, Olsson J, Berglund A, Jones AG (2005) Group stability and homing behaviour but no kin group structure in a coral reef fish. Behav Ecol 16:521–527CrossRefGoogle Scholar
  36. Limbaugh C (1961) Cleaning symbiosis. Sci Am 205:42–49CrossRefGoogle Scholar
  37. Losey GS, Balazs GH, Privitera LA (1994) Cleaning symbiosis between the wrasse, Thalassoma duperrey, and the green turtle, Chelonia mydas. Copeia 3:684–690CrossRefGoogle Scholar
  38. Marcusso PF, Yunis J, Claudiano GS, Manrique WG, Salvador R, de Moraes JRE, Moraes FR (2014) Sodium fluorescein for early detection of skin ulcers in Aeromonas hydrophila infected Piaractus mesopotamicus. Bull Eur Assoc Fish Pathol 34:102–106Google Scholar
  39. McCammon AM, Sikkel P, Nemeth D (2010) Effects of three Caribbean cleaner shrimps on ectoparasitic monogeneans in a semi-natural environment. Coral Reefs 29:419–426CrossRefGoogle Scholar
  40. Militz TA, Hutson KS (2015) Beyond symbiosis: cleaner shrimp clean up in culture. PLoS One 10(2):e0117723CrossRefPubMedPubMedCentralGoogle Scholar
  41. Neiffer DL, Stamper MA (2009) Fish sedation, anesthesia, analgesia, and euthanasia: considerations, methods, and types of drugs. ILAR J 50:343–360CrossRefPubMedGoogle Scholar
  42. Noga EJ, Udomkusonri P (2002) Fluorescein: a rapid, sensitive, nonlethal method for detecting skin ulceration in fish. Vet Pathol 39:726–731CrossRefPubMedGoogle Scholar
  43. Oates J, Manica A, Bshary R (2010) Roving and service quality in the cleaner wrasse Labroides bicolor. Ethology 116:309–315CrossRefGoogle Scholar
  44. Östlund-Nilsson S, Becker JHA, Nilsson GE (2005) Shrimps remove ectoparasites from fishes in temperate waters. Biol Lett 1:454–456CrossRefPubMedPubMedCentralGoogle Scholar
  45. Penning MR, Vaughan DB, Fivas K, McEwan T (2017) Chemical immobilization of elasmobranchs at uShaka Sea World, Durban, South Africa. In: Smith M, Warmolts D, Thoney D, Heuter R, Murray M, Ezcurra J (eds) The elasmobranch husbandry manual II: recent advances in the care of sharks, rays and their relatives. Special Publication of the Ohio Biological Survey, Columbus, Ohio, pp 331–338Google Scholar
  46. Purivirojkul W (2012) Histological changes of aquatic animals by parasitic infection. In: Martinez EP (ed) Histopathology—reviews and recent advances. InTech publishers, London, pp 153–176Google Scholar
  47. R Development Core Team (2017) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Accessed 20 Sept 2017
  48. Roberts RJ (2012) Fish pathology, 4th edn. Wiley, New YorkCrossRefGoogle Scholar
  49. Rohde K (2005) Marine parasitology. CSIRO Publishing, ClaytonCrossRefGoogle Scholar
  50. Sazima I, Grossman A, Sazima C (2004) Hawksbill turtles visit moustached barbers: cleaning symbiosis between Eretmochelys imbricata and the shrimp Stenopus hispidus. Biota Neotrop 4:1–6CrossRefGoogle Scholar
  51. Sazima C, Grossman A, Sazima I (2010) Turtle cleaners: reef fishes foraging on epibionts of sea turtles in the tropical Southwestern Atlantic, with a summary of this association type. Neotrop Ichthyol 8:187–192CrossRefGoogle Scholar
  52. Sazima C, Jordano P, Guimarães PR Jr, Dos Reis SF, Sazima I (2012) Cleaning associations between birds and herbivorous mammals in Brazil: structure and complexity. Auk 129:36–43CrossRefGoogle Scholar
  53. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez J-Y, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682CrossRefPubMedGoogle Scholar
  54. Shapiro DY (1988) Variation of group composition and spatial structure with group size in a sex-changing fish. Anim Behav 36:140–149CrossRefGoogle Scholar
  55. Shephard KL (1994) Functions of fish mucus. Rev Fish Biol Fish 4:401–429CrossRefGoogle Scholar
  56. Sikkel PC, Cheney KL, Côté IM (2004) In situ evidence for ectoparasites as a proximate cause of cleaning interactions in reef fish. Anim Behav 68:241–247CrossRefGoogle Scholar
  57. Soares MC, Bshary R, Cardoso SC, Côté IM (2008) The meaning of jolts by fish clients of cleaning gobies. Ethology 114:209–214CrossRefGoogle Scholar
  58. Soares MC, Oliviera RF, Ros AFH, Grutter AS, Bshary R (2011) Tactile stimulation lowers stress in fish. Nat Commun. PubMedCrossRefGoogle Scholar
  59. Sun D, Cheney KL, Werminghausen J, Meekan MG, McCormick MI, Cribb TH, Grutter AS (2015) Presence of cleaner wrasse increases the recruitment of damselfishes to coral reefs. Biol Lett 11:20150456CrossRefPubMedPubMedCentralGoogle Scholar
  60. Titus BM, Daly M, Exton DA (2015) Temporal patterns of Pederson shrimp (Ancylomenes pedersoni Chace 1958) cleaning interactions on Caribbean coral reefs. Mar Biol 162:1651–1664CrossRefGoogle Scholar
  61. Titus BM, Vondriska C, Daly M (2017) Comparative behavioural observations demonstrate the ‘cleaner’ shrimp Periclimenes yucatanicus engages in true symbiotic cleaning interactions. R Soc Open Sci 4:170078CrossRefPubMedPubMedCentralGoogle Scholar
  62. Vaughan DB, Penning MR, Christison KW (2008) 2-Phenoxyethanol as anaesthetic in removing and relocating 102 species of fishes representing 30 families from Sea World to uShaka Marine World, South Africa. Onderstepoort J Vet Res 75:189–198CrossRefPubMedGoogle Scholar
  63. Vaughan DB, Grutter AS, Costello MJ, Hutson K (2016) Cleaner fishes and shrimp diversity and a re-evaluation of cleaning symbioses. Fish Fish. CrossRefGoogle Scholar
  64. Waldie PA, Blomberg SP, Cheney KL, Goldizen AW, Grutter AS (2011) Long-term effects of the cleaner fish Labroides dimidiatus on coral reef fish communities. PLoS One 6:1–7CrossRefGoogle Scholar
  65. Walsh WJ (1983) Stability of a coral reef fish community following a catastrophic storm. Coral Reefs 2:49–63CrossRefGoogle Scholar
  66. Weeks P (2000) Red-billed oxpeckers: vampires or tickbirds? Behav Ecol 11:154–160CrossRefGoogle Scholar
  67. Wheeler B, Torchiano M (2016) lmPerm: Permutation tests for linear models. R package version 2.1.0. Accessed 20 Sept 2017
  68. Whiteman EA, Côté IM (2002) Cleaning activity of two Caribbean cleaning gobies: intra- and interspecific comparisons. J Fish Biol 60:1443–1458CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Centre for Sustainable Tropical Fisheries and Aquaculture, College of Science and EngineeringJames Cook UniversityTownsvilleAustralia
  2. 2.School of Biological SciencesThe University of QueenslandSt LuciaAustralia
  3. 3.School of Veterinary MedicineSt George’s UniversityGrenadaGrenada
  4. 4.Division of Tropical Health and MedicineJames Cook UniversityTownsvilleAustralia

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