Characterisation of chitin in the cuticle of a velvet worm (Onychophora)

We characterize the trunk cuticle of velvet worms of the Peripatoides novaezealandiae-group (Onychophora) using SEM, TEM, Fourier transform infrared spectroscopy (FT-IR), and thermogravimetric analysis (TGA). TEM and SEM revealed a relatively uniform organization of the delicate cuticle that is covered by numerous bristled and nonbristled papillae with ribbed scales arranged in transverse rows. The cuticle consists of a very thin multilayered epicuticle of varying appearance followed by the largely fibrous procuticle. The irregularly arranged nanofibres of isolated cuticular chitin seen by SEM are considered as bundles of chitin fibres. FT-IR and TGA showed that the chitin is of the α-type. This confirms and broadens the single previous study in which the presence of α-chitin in a velvet worm was demonstrated with a single analysis (X-ray diffraction).

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  • Aguinaldo AM, Tubeville JM, Linford LS, Rivera MC, Garey JR et al. (1976). Evidence for a clade of nematodes, arthropods and other moulting animals. Nature 387: 489-493. doi: 10.1038/387489a0
  • Borner J, Rehm P, Schill RO, Ebersberger I, Burmester T (2014). A transcriptome approach to ecdysozoan phylogeny. Molecular Phylogenetics and Evolution 80: 79-87. doi: 10.1016/j. ympev.2014.08.001
  • Campbell LI, Rota-Stabelli O, Edgecombe GD, Marchioro T, Longhorn SJ et al. (2011). MicroRNAs and phylogenomics resolve the relationships of Tardigrada and suggest that velvet worms are the sister group of Arthropoda. Proceedings of the National Academy of Sciences of the United States of America 108: 15920-15924. doi: 10.1073/pnas.1105499108
  • Cuong HN, Minh NC, Hoa NV, Trung TS (2016). Preparation and characterization of high purity β-chitin from squid pens (Loligo chenisis). International Journal of Biological Macromolecules 93: 442-447. doi: 10.1016/j.ijbiomac.2016.08.085
  • Dunn CW, Hejnol A, Matus DO, Pang K, Browne WE et al. (2008). Broad phylogenomic sampling improves resolution of the animal tree of life. Nature 452: 745-749. doi: 10.1038/ nature06614
  • Greven H, Peters W (1986). Localization of chitin in the cuticle of Tardigrada using wheat germ agglutinin–gold conjugate as a specific electron-dense marker. Tissue and Cell 18: 297-304.
  • Greven H, Kaya M, Baran T (2016). The presence of α-chitin in Tardigrada with comments on chitin in the Ecdysozoa. Zoologischer Anzeiger 264: 11-16. doi.org/10.1016/j. jcz.2016.06.003
  • Greven H, Kaya M, Junker K, Akyuz L, Amemiya C (2019). Characterization of tongue worm (Pentastomida) chitin supports α- rather than β-chitin. Zoologischer Anzeiger 279: 111-115.
  • Hackman RH, Goldberg M (1975). Peripatus: Its affinities and its cuticle. Science 190: 582-583. doi: 10.1126/science.190.4214.582
  • Hejnol A, Obst M, Stamatakis A, Ott M, Rouse GW et al. (2009). Assessing the root of bilaterian animals with scalable phylogenomic methods. Proceedings of the Royal Society London, Series B 276: 4261-4270. doi: 10.1098/rspb.2009.0896
  • Jang MK, Kong BG, Jeong YI, Lee CH, Nah JW (2004). Physicochemical characterization of α-chitin, β-chitin, and γ-chitin separated from natural resources. Journal of Polymer Science A1 42: 3423-3432. doi: 10.1002/pola.20176
  • Jeuniaux C (1982). La chitine dans la règne animal. Bulletin de Société zoologique de France 107: 363-386 (article in French with an abstract in English).
  • Karuppaswamy SA (1977). Occurrence of β-chitin in the cuticle of a Pentastomid Raillietiella gowrii. Experientia 33: 735-736. doi: 10.1007/BF01944158
  • Kaya, M, Baublys V, Sargin I, Satkauskiene I, Paulaskas A et al. (2016). How taxonomic relations affect the physicochemical properties of chitin. Food Biophysics 11: 10-19.
  • Kaya M, Mujtaba M, Ehrlich H, Sakaberria AM, Baran T et al. (2017). On chemistry of γ-chitin. Carbohydrate Polymers 176: 177-186. doi: 10.1016/j.carbpol.2017.08.076
  • Krishnan G (1970). Chemical nature of the cuticle and its mode of hardening in Eoperipatus weldon. Acta Histochemica 37: l-17.
  • Kristensen RM, Neuhaus B (1999). The ultrastructure of the tardigrade cuticle with special attention to marine species. Zoologischer Anzeiger 238: 261-281.
  • Kumirska J, Czerwicka M, Kaczynski Z, Bychowska A, Brzozowski K et al. (2010). Application of spectroscopic methods for structural analysis of chitin and chitosan. Marine Drugs 8: 1567-1636. doi: 10.3390/md8051567
  • Kunike G (1925). Nachweis und Verbreitung organischer Skeletsubstanzen bei Tieren. Zeitschrift für vergleichende Physiologie 2: 233-253 (in German).
  • Lankester ER (1904). The structure and classification of the Arthropoda. Quarterly Journal of Microscopical Science 47: 523-582.
  • Lavall RL, Assis OB, Campana-Filho SP (2007). β-Chitin from the pens of Loligo sp.: Extraction and characterization. Bioresource Technology 98: 2465-2472. doi: 10.1016/j.biortech2006.09.002
  • Lavallard RM (1965). Étude au microscope électronique de l’épithélium tégumentaire chez Peripatus acacioi, Marcus et Marcus. Comptes rendus de l’Académie des sciences Série D, Sciences naturelles 260: 965- 968.
  • Lavallard RM (1972). Recherches sur le paroi tégumentaire et le cycle d’intermue chez Peripatus acacioi, Marcus et Marcus. PhD, l’Université Paris, Paris, France.
  • Liu S, Sun J, Yu L, Zhang C, Li J et al. (2012). Extraction and characterization of chitin from the beetle Holotrichia parallela Motschulsky. Molecules 17: 4604-4611. doi: 10.3390/ molecules17044604
  • Lotmar W, Picken LR (1950). A new crystallographic modification of chitin and its distribution. Experientia 6: 58-59. doi: 10.1007/ BF02174818
  • Mallatt J, Garey JR, Shultz JW (2004). Ecdysozoan phylogeny and Bayesian inference: first use of nearly complete 28S and 18S rRNA gene sequences to classify the arthropods and their kin. Molecular Phylogenetics and Evolution 31: 178-191. doi: 10.1016/j.ympev.2003.07.013
  • Meusemann K, von Reumo BM, Simon S, Roeding F, Strauss S et al. (2010). A phylogenomic approach to resolve the arthropod tree of life. Molecular Biology and Evolution 27: 2451-2464. doi: 10.1093/molbev/msq130
  • Mayer G, Kauschke S, Rüdiger J, Stevenson PA (2013). Neural markers reveal a one-segmented head in tardigrades (water bears). PLOS One 8:e59090. doi: 10.1371/journal.pone.0059090
  • Neuhaus B, Kristensen RM, Peters W (1997a). Ultrastructure of the cuticle of Loricifera and demonstrations of chitin using gold labelled with wheat germ agglutinin. Acta Zoologica Stockholm 78: 215-225. doi: 10.1111/j.1463-6395.1997.tb01008.x
  • Neuhaus B, Bresciani J, Peters W (1997b). Ultrastructure of the pharyngeal cuticle and lectin labelling with wheat germ agglutinin–gold conjugate indicating chitin in the pharyngeal cuticle of Oesophagostomum dentatum (Strongylida, Nematoda). Acta Zoologica Stockholm 78: 205-213.
  • Neville AC (1975). Biology of the Arthropod Cuticle. Berlin, Germany: Springer.
  • Nielsen C (2012). Animal Evolution: Interrelationships of the Living Phyla. 3rd ed. Oxford, UK: University Press. Oliveira IS; Morley V; Read VM St J, Mayer G (2012). A world checklist of Onychophora (velvet worms), with notes on nomenclature and status of names. ZooKeys 211: 1-70. doi: 10.3897/zookeys.211.3463
  • Prasath T, Greven H, D’Haese J (2013). EF-hand proteins in onychophorans as compared to tardigrades and other ecdysozoans. Journal of Limnology 72: 44-53.
  • Pripnow B, Ruhberg H (2003). Peripatopsidae (Onychophora) from New Zealand: observations on selected morphs of the ‘Peripatoides novaezealandiae-complex’ in culture: morphological and reproductive aspects. African Invertebrates 44: 103-114.
  • Robson EA (1964). The cuticle of Peripatopsis moseleyi. Quarterly Journal of Microscopical Science 105: 281-299.
  • Rota-Stabelli O, Kayal E, Gleeson D, Daub J, Boore J et al. (2010). Ecdysozoan mitogenomics: Evidence for a common origin of the legged invertebrates, the Panarthropoda. Genome Biology and Evolution 2: 425-440. doi: 10.1093/gbe/evq030
  • Rudall KM (1955). The distribution of collagen and chitin. Symposia of the Society for Experimental Biology 9: 49-70.
  • Rudall KM, Kenchington W (1973). The chitin system. Biological Reviews 49: 597-610. doi: 10.1111/j.1469-185X.1973.tb01570.x
  • Schmidt-Rhaesa A, Bartolomaeus Th, Lemburg Ch, Ehlers U, Garey J (1998). The position of the Arthropoda in the phylogenetic system. Journal of Morphology 238: 263-285. doi: 10.1002/ (SICI)1097-4687(199812)238:3<263
  • Telford MJ, Bourlat SJ, Economou A, Papillon D, Rota-Stabelli O (2008). The evolution of the Ecdysozoa. Philosophical Transactions of the Royal Society B 363: 1529-1537. doi: 10.1098/rstb.2007.2243
  • Trewick SA (2000). Mitochondrial DNA sequences support allozyme evidence for cryptic radiation of New Zealand Peripatoides (Onychophora). Molecular Ecology 9: 269-281. doi: 10.1046/j.1365-294x.2000.00873
  • Wang Y, Chang Y, Yu L, Zhang C, Xu X et al. (2013). Crystalline structure and thermal property characterization of chitin from Antarctic krill (Euphausia superba). Carbohydrate Polymers 92: 90-97. doi: 10.1016/j.carbpol.2012.09.084
  • Vincent JFV, Wegst GK (2004). Design and mechanical properties of insect cuticle. Arthropod Structure and Development 3: 187- 199. doi: 10.1016/j.asd.2004.05.006
  • Westheide W, Rieger G (2013). Spezielle Zoologie. Teil 1: Einzeller und Wirbellose Tiere. 3rd ed. Heidelberg, Germany: SpringerSpektrum (in German).
  • Wright JC, Luke BM (1989). Ultrastructural and histochemical investigations of Peripatus integument. Tissue and Cell 21: 605-625.
  • Zhang M, Haga A, Sekiguchi, H, Hirano S (2000). Structure of insect chitin isolated from beetle larva cuticle and silkworm (Bombyx mori) pupa exuvia. International Journal of Biological Macromolecules 27: 99-105. doi: 10.1016/S0141- 8130(99)00123-3.