New perspectives in a microstructure study of the bovine and bubaline claw

Assis BM, Borges LPB, Pereira KF, de Oliveira Lima CR, da Silva LAF, Teixeira PPM, Rabelo RE (2022): New perspectives in a microstructure study of the bovine and bubaline claw. Vet Med-Czech 67, 395–407.

download PDF

The techniques of microtomography (Micro-CT), confocal laser scanner microscopy (CLSM), atomic force microscopy (AFM), nanoindentation – Vickers hardness (Nano-VH) and X-ray fluorescence (XRF) are undeniably important to the modern study of bovine podiatry. These techniques are also employed in engineering, physics and in the assessment of biomaterials used in reconstructive or experimental surgeries in bovine and bubaline claws. Although studies involving these analyses are still inconspicuous in veterinary medicine, these technologies represent a new paradigm in this area, enabling the development of new lines of research. The objective of this review is to gather information about the microstructural aspects of bovine and bubaline claws, concerning the intratubular and extratubular keratin, which is responsible for the physical and mechanical structure of the claw capsule. This study elucidates different methods used to evaluate the hooves of healthy and sick animals through a micrometric analysis and nano-scale analyses. We would like to emphasise that the described techniques can be applied to study other species.

Abdullahi U, Salihi A, Ali AB. Hardness behaviour of thermoplastic cattle horn using nanoindentation technique. Asian J Engineer Technol. 2014 Apr;2(2):169-73.
Alsteens D, Dupres V, Yunus S, Latge JP, Heinisch JJ, Dufrene YF. High-resolution imaging of chemical and biological sites on living cells using peak force tapping atomic force microscopy. Langmuir. 2012 Dec 11;28(49):16738-44.
Andrade JP, Mercurio DG, Campos PMBGM. Avaliacao celular das estruturas cutaneas por meio da microscopia confocal de reflectancia [Evaluation of cutaneous cellular structures by reflectance confocal microscopy]. RBM Rev Bras Med. 2015;71:5-14. Portugues.
Assis BM. Histomorfometria, microtomografia bidimensional e tridimensional, teste de nanodureza e composicao bioquimica do estojo corneo de bubalinos [Histomorphometry, bidimensional and tridimensional microtomography, nanohardness test and biochemical composition of hoof capsule of buffaloes] [PhD thesis]. Goiânia, Goiás: Universidade Federal de Goiás; 2015. 81 p. Portuguese.
Assis BM, Silva LAF, Lima CRO, Sant’Ana FJF, Santos GP, Vulcani VAS, Rabelo RE. Microtomographic parameters and nanoindentation of the hoof of Girolando cattle. Anat Histol Embryol. 2017a Oct;46(5):456-63.
Assis BM, Silva LAF, Lima CRO, Gouveia RF, Vulcani VAS, de Sant’Ana FJF, Rabelo RE. Microstructure and hardness of buffalo’s hoofs. Anat Histol Embryol. 2017b Oct;46(5):439-45.
Assis BM, Vulcani VAS, Silva LAF, Dias M, Pancotti A, Lima CRO, Rabelo RE. Biochemical composition of the hoof capsule of buffaloes and its influence on hoof quality. Arq Bras Med Vet Zootec. 2017c Feb;69(1):57-64.
Bajanowski T, Kohler H, Schmidt PF, von Saldern CF, Brinkmann B. The cloven hoof in legal medicine. Int J Legal Med. 2001;114(6):346-8.
Bragulla HKD, Budras G, Mulling S, HE Konig. Forelimb or thoracic limb (membra thoracica). In: Konig HE, Bragulla HKD, editors. Veterinary anatomy of domestic animals. Stuttgart: Schattauer Verlag GmbH; 2004. p. 1-48.
Bruker. Bruker microCT – User manual. Desk-top X-ray microtomograph SkyScan 1272 [Internet]. 2013 [cited 2020 Mar 2]. Available from:
Calistu AE, Cara IG, Topa D, Jitareanu G. Analyzing soil porosity under different tillage systems using X-ray microtomography. Lucr Stiint (Ser Agron). 2016;59(1):201-4.
de Carvalho MPN, Sant’Anna SS, Grego KF, de Campos Fonseca-Pinto ACB, Lorigados CAB, Queiroz-Hazarbassanov NGT, Catao-Dias JL. Microcomputed tomographic, morphometric, and histopathologic assessment of congenital bone malformations in two neotropical viperids. J Wildl Dis. 2017 Oct;53(4):804-15.
Coelho PG, Giro G, Kim W, Granato R, Marin C, Bonfante EA, Bonfante S, Lilin T, Suzuki M. Evaluation of collagen-based membranes for guided bone regeneration, by three-dimensional computerized microtomography. Oral Surg Oral Med Oral Pathol Oral Radiol. 2012 Oct;114(4):437-43.
Cruz AF, Queiroz PJB, Assis BM, Silva DC, Rabelo RE, Silva LAF. [Use of atomic force microscopy (AFM) to evaluate the surface microstructure of the abaxial wall of the hoof of calves supplemented with biotin – Partial results]. In: II Encontro Cientifico da Escola de Veterinaria e Zootecnia da Universidade Federal de Goias, 2017, Goiânia-GO. Anais do II Encontro Científico da Escola de Veterinária e Zootecnia da Universidade Federal de Goiás. Goiânia-GO: Escola de Veterinária e Zootecnia da Universidade Federal de Goiás; 2017; p. 174-9. Portuguese.
Duajie L. Manual, a better mensure. Textile texture measurement using 3D profilometry [Internet]. 2017 [cited 2020 Mar 2]. Available from:
Dufrene YF, Martinez-Martin D, Medalsy I, Alsteens D, Muller DJ. Multiparametric imaging of biological systems by force-distance curve-based AFM. Nat Methods. 2013 Sep;10(9):847-54.
Fischer-Cripps AC. Nanoindentation – Hardcover. New York: Springer International Publishing; 2011. 282 p.
Fontelles MJ, Simoes MG, Almeida JC, Fontelles RGC. Metodologia da pesquisa: Diretrizes para o calculo do tamanho da amostra [Scientific research methodology: Guidelines for size sample calculation]. Rev Para Med. 2010;24(2):57-64. Portuguese.
Freitas SLR. Venografia dos digitos de animais jovens antes e apos aplicacao intrarruminal de oligofrutose [Venographic study of foot of young bovine before and after intrarruminal oligofructose application] [Master thesis]. Goiânia, Goiás: Universidade Federal de Goiás, Escola de Veterinária e Zootecnia; 2015. Portuguese.
Ghassemi-Armaki H, Maaß R, Bhat SP, Sriram S, Greer JR, Kumar KS. Deformation response of ferrite and martensite in a dual-phase steel. Acta Mater. 2014;62:197-211.
Greenough PR. Bovine laminitis and lameness – A hands on approach. Philadelphia: Sounders Elsevier; 2007. 311 p.
He BB, Huang MX. Revealing the intrinsic nanohardness of lath martensite in low carbon steel. Metall Mater Trans A Phys Metall Mater Sci. 2015;46(2):688-94.
Kasapi MA, Gosline JM. Design complexity and fracture control in the equine hoof wall. J Exp Biol. 1997 Jun;200(Pt 11):1639-59.
Kung M. Die Zugfestigkeit des Hufhorns von Pferden. Messungen an definierten Stellen des Hufes sowie unter dem Einfluss verschiedener Umgebungsbedingungen [Tensile strength of the hoof horn of horses. Measurements at defined points on the hoof and under the influence of various environmental conditions] [PhD thesis]. Switzerland: University of Zürich; 1991. German.
LNNano – Laboratório Nacional de Nanotecnologia. Microscopia a laser confocal (perfiolometria) [Internet]. 2017 [cited 2020 May 2]. Available from:
Lima I, Appoloni C, Oliveira LC, Lopes RT. Caracterizacao de materiais ceramicos atraves da microtomografia computadorizada 3D [Characterization of ceramic materials through 3D computerized microtomography]. Rev Bras Arqueometria, Restaur Conserv. 2007;1(2):22-7. Portuguese.
Lima I, Lopes RT, Oliveira LF, Alves JM. Analise de estrutura ossea atraves de microtomografia computadorizada 3D [Bone structure analysis using 3D computed microtomography]. Rev Bras Fís Méd. 2009;2(1):6-10. Portuguese.
Lopes AP, Fiori AP, Reis Neto JM, Marchese C, Vasconcellos EMG, Trzascos B, Onishi CT, Pinto-Coelho CV, Secchi R, Silva GF. Analise tridimensional de rochas por meio de microtomografia computadorizada de raios-x integrada a petrografia [X-ray microtomography as tool for microstructural and mineral phase analysis]. Geociencias. 2012;31(1):129-42. Portuguese.
Loureiro MG. Estudo da tecnica de venografia dos digitos de vacas [Study of venography technique in digits of cows] [PhD thesis]. Botucatu: Universidade Estadual Paulista, Faculdade de Medicina Veterinária e Zootecnia; 2013. Portugues.
Lovric G, Barre SF, Schittny JC, Roth-Kleiner M, Stampanoni M, Mokso R. Dose optimization approach to fast X-ray microtomography of the lung alveoli. J Appl Crystallogr. 2013 Aug 1;46(Pt 4):856-60.
Mendes FBR, Castro RPR, Macedo MP, Pinto CAL, Duprat Neto JP, Rezze GG. Diagnostico de pseudomelanoma na microscopia confocal: O desafio das celulas dendriticas epidermicas [Pseudomelanoma diagnostic through confocal microscopy: The challenge of the epidermal dendritic cells]. Surg Cosmet Dermatol. 2015;7(3):246-8. Portuguese.
Metscher BD. Biological applications of X-ray microtomography: Imaging micro-anatomy, molecular expression and organismal diversity. Microsc Anal (Am Ed). 2013 Mar;27(2):13-6.
Miller KJ, Zhua W, Laurent GJ, Glenn M, Gaetanib A. Experimental quantification of permeability of partially molten mantle rock. Earth Planet Sci Lett. 2014;388(2):273-82.
Nanosurf FlexAFM. Manual operating instructions for C3000 control software version 3.6. Switzerland; 2017. 332 p.
Queiroz PJB, Assis BM, Silva DC, Noronha Filho ADF, Pancotti A, Rabelo RE, Borges NC, Vulcani VAS, Silva LAFD. Mineral composition and microstructure of the abaxial hoof wall in dairy heifers after biotin supplementation. Anat Histol Embryol. 2021 Jan;50(1):93-101.
Rabelo RE, Vulcani VAS, Sant’Ana FJF, Silva LAF, Assis BM, Araujo GHM. Microstructure of Holstein and Gir breed adult bovine hooves: Histomorphometry, three-dimensional microtomography and microhardness test evaluation. Arq Bras Med Vet Zootec. 2015 Nov-Dec;67(6):1492-500.
Rebelo LM, de Sousa JS, Mendes Filho J, Radmacher M. Comparison of the viscoelastic properties of cells from different kidney cancer phenotypes measured with atomic force microscopy. Nanotechnology. 2013 Feb 8;24(5):1-12.
Regnault S, Hutchinson JR, Jones ME. Sesamoid bones in tuatara (Sphenodon punctatus) investigated with X-ray microtomography, and implications for sesamoid evolution in Lepidosauria. J Morphol. 2017 Jan;278(1):62-72.
Renghini C, Giuliani A, Mazzoni S, Brun F, Larsson E, Baino F, Vitale-Brovarone C. Microstructural characterization and in vitro bioactivity of porous glass-ceramic scaffolds for bone regeneration by synchrotron radiation X-ray microtomography. J Eur Ceram Soc. 2013;33(9):1553-65.
Roche B, David V, Vanden-Bossche A, Peyrin F, Malaval L, Vico L, Lafage-Proust MH. Structure and quantification of microvascularisation within mouse long bones: What and how should we measure? Bone. 2012 Jan;50(1):390-9.
Santhiago M, Bettini J, Araujo SR, Bufon CC. Three-dimensional organic conductive networks embedded in paper for flexible and foldable devices. ACS Appl Mater Interfaces. 2016 May 4;8(17):10661-4.
Santhiago M, Strauss M, Pereira MP, Chagas AS, Bufon CC. Direct drawing method of graphite onto paper for high-performance flexible electrochemical sensors. ACS Appl Mater Interfaces. 2017 Apr 5;9(13):11959-66.
Santos AC, Mendes CMC, Rosa FP. A microtomografia computadorizada aplicada a bioengenharia tecidual ossea com o uso de biomateriais [The X-ray microcomputed tomography applied to bioengineering bone tissue with the use of biomaterials]. Rev Cienc Med Biol, Salvador. 2013;12(4):478-81. Portuguese.
Sargentini C, Tocci R, Andrenelli L, Giorgetti A. Preliminary studies on hoof characteristics in Amiata donkey. Ital J Anim Sci. 2012;11(1):123-7.
Schaller O. Nomenclatura anatomica veterinaria ilustrada [Illustrated veterinary anatomical nomenclature]. São Paulo: Editora Manole Ltda; 1999. 614 p.
Silva LH. Morfometria radiografica e tomografica em digitos de bovinos e bubalinos [Radiographic and tomographic morphometry in digiti of the cattle and buffaloes] [Master thesis]. Goiânia, Goiás: Universidade Federal de Goiás, Escola de Veterinária e Zootecnia; 2012. Portuguese.
Tesarova M, Zikmund T, Kaucka M, Adameyko I, Jaros J, Palousek D, Skaroupka D, Kaiser J. Use of micro computed-tomography and 3D printing for reverse engineering of mouse embryo nasal capsule. J Instrum. 2016 Mar;11(3):1-11.
Wagner C, Fournier N, Tautz FS, Temirov R. Measurement of the binding energies of the organic-metal perylene-teracarboxylic-dianhydride/Au111 bonds by molecular manipulation using an atomic force microscope. Phys Rev Lett. 2012 Aug 17;109(7):076102.
Wang QH, Kalantar-Zadeh K, Kis A, Coleman JN, Strano MS. Electronics and optoelectronics of two-dimensional transition metal dichalcogenides. Nat Nanotechnol. 2012 Nov;7(11):699-712.
Zamiri M, Anwar F, Klein BA, Rasoulof A, Dawson NM, Schuler-Sandy T, Deneke CF, Ferreira SO, Cavallo F, Krishna S. Antimonide-based membranes synthesis integration and strain engineering. Proc Natl Acad Sci U S A. 2017 Jan 3;114(1):1-8.
Zhu W, Gaetani GA, Fusseis F, Montesi LG, De Carlo F. Microtomography of partially molten rocks: Three-dimensional melt distribution in mantle peridotite. Science. 2011 Apr 1;332(6025):88-91.
download PDF

© 2022 Czech Academy of Agricultural Sciences | Prohlášení o přístupnosti