, Volume 100, Issue 11, pp 1051–1059 | Cite as

Nest sanitation through defecation: antifungal properties of wood cockroach feces

  • Rebeca B. Rosengaus
  • Kerry Mead
  • William S. Du Comb
  • Ryan W. Benson
  • Veronica G. Godoy
Original Paper


The wood cockroach Cryptocercus punctulatus nests as family units inside decayed wood, a substrate known for its high microbial load. We tested the hypothesis that defecation within their nests, a common occurrence in this species, reduces the probability of fungal development. Conidia of the entomopathogenic fungus, Metarhizium anisopliae, were incubated with crushed feces and subsequently plated on potato dextrose agar. Relative to controls, the viability of fungal conidia was significantly reduced following incubation with feces and was negatively correlated with incubation time. Although the cockroach's hindgut contained abundant β-1,3-glucanase activity, its feces had no detectable enzymatic function. Hence, these enzymes are unlikely the source of the fungistasis. Instead, the antifungal compound(s) of the feces involved heat-sensitive factor(s) of potential microbial origin. When feces were boiled or when they were subjected to ultraviolet radiation and subsequently incubated with conidia, viability was “rescued” and germination rates were similar to those of controls. Filtration experiments indicate that the fungistatic activity of feces results from chemical interference. Because Cryptocercidae cockroaches have been considered appropriate models to make inferences about the factors fostering the evolution of termite sociality, we suggest that nesting in microbe-rich environments likely selected for the coupling of intranest defecation and feces fungistasis in the common ancestor of wood cockroaches and termites. This might in turn have served as a preadaptation that prevented mycosis as these phylogenetically related taxa diverged and evolved respectively into subsocial and eusocial organizations.


Metarhizium anisopliae Termites Preadaptation Cryptocercus punctulatus β-1,3-Glucanases Sociality 



We thank Drs. John Wenzel, Nan-Yao Su and Thomas Chouvenc for providing Cryptocercus cockroaches. Ashley McGuire helped with the DAPI technique. Dr. Christine Nalepa provided insightful comments during the preparation of this manuscript. We also thank four anonymous reviewers for their helpful comments and suggestions. This work was partially funded by an NSF (DEB 0447316) REU grant to RB Rosengaus and by a Northeastern University TIER1 award to RB Rosengaus, V Godoy, E Cram and M Hincapie. All colonies were contained in a USDA inspected and approved facility. The authors declare that they have no conflict of interest. These experiments comply with the current laws of the United States of America and were conducted under USDA permits.

Author contributions

RBR conceived and designed the experiment, analyzed the data, and wrote the manuscript. KM performed all the germination enumeration and DAPI staining. RBR together with WD, RB, and VG ran the glucanase gels and GDL experiments, while RB was involved in running all filtration experiments.

Supplementary material

114_2013_1110_MOESM1_ESM.docx (181 kb)
ESM 1 (DOCX 180 kb)


  1. Batra LR, Batra SWT (1966) Fungus-growing termites of tropical India and associated fungi. J Kansas Entomol Soc 39:725–738Google Scholar
  2. Batra LR, Batra SWT, Bohart GE (1973) The mycoflora of domesticated and wild bees (Apoidea). Mycopathol Mycol Appl 49:13–44CrossRefGoogle Scholar
  3. Bell WJ, Roth LM, Nalepa CA (2007) Cockroaches: ecology, behavior, and natural history. Johns Hopkins University Press, BaltimoreGoogle Scholar
  4. Berlanga M, Paster BJ, Ricardo Guerrero R (2009) The taxophysiological paradox: changes in the intestinal microbiota of the xylophagous cockroach Cryptocercus punctulatus depending on the physiological state of the host. Int Microbiol 12:227–236PubMedGoogle Scholar
  5. Bowman SM, Free SJ (2006) The structure and synthesis of the fungal cell wall. Bioessays 28(8):799–808PubMedCrossRefGoogle Scholar
  6. Bragatto I, Genta FA, Ribeiro AF, Terra WR, Ferreira C (2010) Characterization of a β-1,3-glucanase active in the alkaline midgut of Spodoptera frugiperda larvae and its relation to β-glucan-binding proteins. Insect Biochem Mol Biol 40:861–872PubMedCrossRefGoogle Scholar
  7. Brossut R, Sreng L (1985) L’univers chimique des blattes. Bull Soc Entomologique de Fr 90:1266–1280Google Scholar
  8. Bulmer MS, Bachelet I, Raman R, Rosengaus RB, Sasisekharan R (2009) Targeting an antimicrobial effector function in insect immunity as a pest control strategy. Proc Natl Acad Sci U S A 106:12652–12657PubMedCrossRefGoogle Scholar
  9. Bulmer MS, Denier D, Velenovsky J, Hamilton C (2012) A common antifungal defense strategy in Cryptocercus woodroaches and termites. Insect Soc 59(4):469–478CrossRefGoogle Scholar
  10. Burnham L (1978) Survey of social insects in the fossil record. Psyche 85:85–133CrossRefGoogle Scholar
  11. Chouvenc T, Su NY, Robert A (2009) Inhibition of Metarhizium anisopliae in the alimentary tract of the eastern subterranean termite Reticulitermes flavipes. J Invertebr Pathol 101(2):130–136. doi: 10.1016/j.jip.2009.04.005 Google Scholar
  12. Chouvenc T, Su NY, Robert A (2010) Inhibition of the fungal pathogen Metarhizium anisopliae in the alimentary tracts of five termite (Isoptera) species. Florida Entomol 93(3):467–469. doi: 10.1653/024.093.0327 Google Scholar
  13. Chouvenc T, Efstathion CA, Elliott ML, Su NY (2012) Resource competition between two fungal parasites in subterranean termites. Naturwissenschaften 99(11):949–958PubMedCrossRefGoogle Scholar
  14. Chouvenc T, Efstathion CA, Elliott ML, Su NY (2013) Extended disease resistance emerging from the faecal nest of a subterranean termite. Proc R Soc B 280:20131885. doi: 10.1098/rspb.2013.1885
  15. Cleveland LR, Hall SR, Sanders EP, Collier J (1934) The wood-feeding roach, Cryptocercus, its protozoa and the symbiosis between protozoa and roach. Mem Am Acad Arts Sci 17:185Google Scholar
  16. Currie CR, Wong B, Stuart AE, Schultz TR, Rehner SA, Mueller UG, Sung GH, Spatafora JW, Straus NA (2003) Ancient tripartite coevolution in the attine ant-microbe symbiosis. Science 299:386–388PubMedCrossRefGoogle Scholar
  17. Dillon RJ, Charnley AK (1986) Invasion of the pathogenic fungus Metarhizium anisopliae through the guts of germfree desert locusts Schistocerca gregaria. Mycopathlogia 96:59–66CrossRefGoogle Scholar
  18. Dillon RJ, Charnley AK (1995) Chemical barriers to gut infection in the desert locust: in vivo production of antimicrobial phenols associated with the bacterium Pantoea agglomerans. J Invertebr Pathol 66:72–75CrossRefGoogle Scholar
  19. Genta FA, Bragatto I, Terra WR, Ferreira C (2009) Purification, characterization and sequencing of the major β-1,3-glucanase from the midgut of Tenebrio molitor larvae. Insect Biochem Mol Biol 39:861–874PubMedCrossRefGoogle Scholar
  20. Grandcolas P (1997). What did the ancestors of the woodroach Cryptocercus look like? A phylogenetic study of the origin of subsociality in the subfamily Polyphaginae (Dictyoptera, Blattaria). In: P. Grandcolas (ed) The origin of biodiversity in insects: phylogenetic tests of evolutionary scenarios. Paris, Mémoires du Muséum national d'Histoire Naturelle 173:231–252Google Scholar
  21. Grandcolas P, Deleporte P (1996) The origin of protistan symbionts in termites and cockroaches: a phylogenetic analysis. Cladistics 12:93–98CrossRefGoogle Scholar
  22. Grassé PP (1986) Termintologia vol III. Masson, ParisGoogle Scholar
  23. Haine ER (2007) Symbiont-mediated protection. Proc R Soc Ser B 275:353–361CrossRefGoogle Scholar
  24. Hamilton WD (1978) Evolution and diversity under bark. In: Mound LA, Waloff N (eds) Diversity of insect faunas. Halsted Press, New York, pp 154–175Google Scholar
  25. Hamilton C, Lay F, Bulmer MS (2011) Subterranean termite prophylactic secretions and external antifungal defenses. J Insect Physiol 57(9):1259–1266PubMedCrossRefGoogle Scholar
  26. Hulcr J, Adams AS, Raffa K, Hofstetter RW, Klepzig KD, Currie CR (2011) Presence and diversity of Streptomyces in Dendroctonus and sympatric bark beetle galleries across North America. Microb Ecol 61(4):759–768PubMedCrossRefGoogle Scholar
  27. Jackson DE, Hart AG (2009) Does sanitation facilitate sociality? Anim Behav 77:e1–e5CrossRefGoogle Scholar
  28. Kaltenpoth M (2009) Actinobacteria as mutualists: general healthcare for insects? Trends Microbiol 17:529–535PubMedCrossRefGoogle Scholar
  29. Kaltenpoth M, Goettler W, Dale C, Stubblefield JW, Herzner G, Roeser-Mueller K, Strohm E (2006) Candidatus Streptomyces philanthi, an endosymbiotic Streptomycete in the antennae of Philanthus digger wasps. Int J Syst Evol Microbiol 56:1403–1411PubMedCrossRefGoogle Scholar
  30. Kirkendall L, Kent D, Raffa K (1997) Interactions among males, females and offspring in bark and ambrosia beetles: the significance of living in tunnels for the evolution of social behavior. In: Choe JC, Crespi BJ (eds) The evolution of social behavior in insects and arachnids. Cambridge University Press, Cambridge, pp 181–215Google Scholar
  31. Klass KD, Nalepa C, Lo N (2008) Review wood-feeding cockroaches as models for termite evolution (Insecta: Dictyoptera): Cryptocerus vs. Parasphaeria boleiriana. Mol Phylogenet Evol 46:809–817PubMedCrossRefGoogle Scholar
  32. Kümmerer K (2009) Antibiotics in the aquatic environment—a review—part I. Chemosphere 75:417–434PubMedCrossRefGoogle Scholar
  33. Lo N, Eggleton P (2011) Termite phylogenetics and co-cladogenesis with symbionts. In: Bignell DE, Roisin Y, Lo N (eds) Biology of Termites: A Modern Synthesis. Springer, Heidelberg, pp 27–50Google Scholar
  34. Lo N, Tokuda G, Watanabe H, Rose H, Slaytor M, Maekawa K, Bandi C, Noda H (2000) Evidence from multiple gene sequences indicates that termites evolved from wood-feeding cockroaches. Curr Biol 10:801–804PubMedCrossRefGoogle Scholar
  35. Mathew GM, Ju YM, Lai CY, Mathew DC, Huang CC (2012) Microbial community analysis in the termite gut and fungus comb of Odontotermes formosanus: the implication of Bacillus as mutualists. FEMS Microbiol Ecol 79(2):504–517PubMedCrossRefGoogle Scholar
  36. Moran NA (2006) Symbiosis. Curr Biol 16:R866–R871PubMedCrossRefGoogle Scholar
  37. Nalepa CA (1984) Colony composition, protozoan transfer and some life history characteristics of the woodroach Cryptocercus punctulatus. Behav Ecol Sociobiol 14(4):273–279CrossRefGoogle Scholar
  38. Nalepa CA (1994) Nourishment and the evolution of termite eusociality. In: Hunt JH, Nalepa CS (eds) Nourishment and the evolution of insect societies. Westview Press, Boulder, pp 57–104Google Scholar
  39. Nalepa CA (2011) Altricial development in wood-feeding cockroaches: the key antecedent of termite eusociality. In: Bignell DE, Roisin Y, Lo N (eds) Biology of termites: a modern synthesis. Springer, New York, pp 69–95Google Scholar
  40. Nalepa CA, Bignell DE, Bandi C (2001) Detritivory, coprophagy, and the evolution of digestive mutualisms in Dictyoptera. Insect Soc 48(3):194–201CrossRefGoogle Scholar
  41. Ohkuma M (2006) Symbiosis in the termite gut. In: Seckbach J (ed) Symbiosis: mechanisms and model systems. Cellular origin, life in extreme habitats and astrobiology, volume 4. Springer, New York, pp 715–730Google Scholar
  42. Ohkuma M, Noda S, Hongoh Y, Nalepa CA, Inoue T (2009) Inheritance and diversification of symbiotic trichonymphid flagellate from a common ancestor of termites and the cockroach Cryptocercus. Proc R Soc Biol Sci 276(1655):239–245CrossRefGoogle Scholar
  43. Olagbemiro TO, Lajide L, Sani KM, Staddon BW (1988) 2-Hydroxy-5-methyl-1,4-benzoquinone from the salivary gland of the soldier termites Odontotermes magdalenae. Experientia 44:1022–1025CrossRefGoogle Scholar
  44. Oliver K, Russell J, Moran N, Hunter M (2003) Facultative bacterial symbionts in aphids confer resistance to parasitic wasps. Proc Natl Acad Sci U S A 100:1803–1807PubMedCrossRefGoogle Scholar
  45. Ottesen EA, Leadbetter JR (2010) Diversity of formyltetrahydrofolate synthetases in the guts of the wood-feeding cockroach Cryptocercus punctulatus and the omnivorous cockroach Periplaneta americana. Appl Environ Microbiol 76(14):4909–4913PubMedCrossRefGoogle Scholar
  46. Pauchet Y, Freitak D, Heidel-Fischer HM, Heckel DG, Voge H (2009) Immunity or digestion: glucanase activity in a glucan-binding protein family from Lepidoptera. J Biol Chem 284:2214–2224PubMedCrossRefGoogle Scholar
  47. Ramsay JA (1971) Insect rectum. Phil Trans R Soc Lond Ser B 262(842):251–260CrossRefGoogle Scholar
  48. Roberts DW, St Leger RJ (2004) Metarhizium spp., cosmopolitan insect-pathogenic fungi: mycological aspects. Adv Appl Microbiol 54:1–70PubMedGoogle Scholar
  49. Rosengaus RB, Guldin MR, Traniello JFA (1998) Inhibitory effect of termite fecal pellets on fungal spore germination. J Chem Ecol 24(10):1697–1706CrossRefGoogle Scholar
  50. Rosengaus RB, Moustakas JE, Calleri DV, Traniello JFA (2003) Nesting ecology and cuticular microbial loads in dampwood (Zootermopsis angusticollis) and drywood termites (Incisitermes minor, I. schwarzi, Cryptotermes cavifrons). J Insect Sci 3:31 (available on Scholar
  51. Rosengaus RB, Traniello JFA, Bulmer MS (2011) Ecology, behavior and evolution of disease resistance in termites. In: Bignell DE, Roisin Y, Lo N (eds) Biology of termites: a modern synthesis. Springer-Verlag, Berlin, pp 165–192Google Scholar
  52. Scarborough CL, Ferrari J, Godfray HCJ (2005) Aphid protected from pathogen by endosymbiont. Science 310:3781CrossRefGoogle Scholar
  53. Schauer C, Thompson CL, Brune A (2012) The bacterial community in the gut of the cockroach Shelfordella lateralis reflects the close evolutionary relatedness of cockroaches and termites. Appl Environ Microbiol 78(8):2758–2767PubMedCrossRefGoogle Scholar
  54. Schluns EA, Wegener BJ, Schluns H, Azuma N, Robson SKA, Crozier RH (2009) Breeding system, colony and population structure in the weaver ant Oecophylla smaragdina. Mol Ecol 18:156–167PubMedCrossRefGoogle Scholar
  55. Schultheis KF (2009) Symbiont-mediated protection against fungal infection in the dampwood termite, Zootermopsis angusticollis. Masters Thesis, Northeastern University, BostonGoogle Scholar
  56. Taylor TN, Hass H, Kerp H, Krings M, Hanlin RT (2005) Perithecial ascomycetes from the 400 million year old Rhynie chert: an example of ancestral polymorphism. Mycologia 97:269–285PubMedCrossRefGoogle Scholar
  57. Thiele-Bruhn S, Peters D (2007) Photodegradation of pharmaceutical antibiotics on slurry and soil surfaces. Landbauforsch 1(57):13–23Google Scholar
  58. Thomashow LS, Bonsall RF, Weller DM (2008) Antibiotic production by soil and rhizosphere microbes in situ. In: Karlovsky P (ed) Secondary metabolites in soil ecology: soil biology. Springer, Heidelberg, pp 23–36CrossRefGoogle Scholar
  59. Thorne BL, Carpenter JM (1992) Phylogeny of the Dictyoptera. Syst Entomol 17:253–268CrossRefGoogle Scholar
  60. Tunaz H, Stanley D (2009) An immunological axis of biocontrol: infections in field-trapped insects. Naturwissenschaften 96(9):1115–1119PubMedCrossRefGoogle Scholar
  61. Weiss MR (2006) Defecation behavior and ecology of insects. Ann Rev Entomol 51:631–661CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Rebeca B. Rosengaus
    • 1
  • Kerry Mead
    • 1
  • William S. Du Comb
    • 2
  • Ryan W. Benson
    • 2
  • Veronica G. Godoy
    • 2
  1. 1.Department of Marine and Environmental SciencesNortheastern UniversityBostonUSA
  2. 2.Department of BiologyNortheastern UniversityBostonUSA

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