Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals

Eastern grey kangaroo (Macropus giganteus) myofibres. 1. A simplified classification method using two commercially available antibodies

N. B. Spiegel A B E , W. H. Johns C , S. D. Sinclair C , P. C. Wynn C D , J. M. Thompson A and P. L. Greenwood C F

A School of Environmental and Rural Science, University of New England, Armidale, NSW 2351, Australia.

B Faculty of Veterinary Science, University of Sydney, Sydney, NSW 2006, Australia.

C Industry & Investment NSW, University of New England, Armidale, NSW 2351, Australia.

D E. H. Graham Centre for Agricultural Innovation, Charles Sturt University, Wagga Wagga, NSW 2678, Australia.

E Present address: School of Environmental Science, Murdoch University, Murdoch, WA 6150, Australia.

F Corresponding author. Email:

Animal Production Science 50(6) 386-392
Submitted: 11 December 2009  Accepted: 14 April 2010   Published: 11 June 2010


Skeletal muscles from eastern grey kangaroos (Macropus giganteus) were assessed for myofibre contractile and metabolic characteristics using immunocytochemical and histological staining of serial sections. Myofibre classification using monoclonal antibodies that typically bind to mammalian slow (clone WB-MHC), fast (clone MY-32) and Types 1, 2X and 2B (clone S5 8H2) myosin heavy chains was validated using acid- and alkali-preincubated myofibrillar ATPase, NADH and α-glycerophosphate dehydrogenase stains. Myofibres were classified as Type 1 (slow oxidative), Type 2A (fast oxidative-glycolytic), Type 2X/2B (fast glycolytic) or intermediate or transitional myofibre Types 2C (Type 1–Type 2A intermediate) and 2AX/B (Type 2A–Type 2X/2B intermediate). The Type 2 (fast) antibody (clone MY-32) used in the present study did not bind to a subset of fast myofibres in any of the eight kangaroo muscles investigated. These myofibres were identified as Type 2A using clone S5 8H2 and on the basis of the histochemical staining profile. Hence, a simplified immunostaining system using only clones WB-MHC (anti-Type 1) and MY-32 (anti-Type 2X/2B) allowed five myofibre types to be identified based on the staining pattern and intensity of staining for the two antibodies. It is concluded that the myofibres of muscles from kangaroos can be quickly classified into five types using two commercially available antibodies. This method is directly applicable for routine investigations into the myofibre properties of commercially important muscles from the kangaroo musculature and, when combined with enzymatic assays for oxidative and glycolytic activity, will allow for a better understanding of factors influencing the quality of meat from kangaroos.

Additional keyword: macropod.


Anapol F Herring SW 2000 Ontogeny of histochemical fibre types and muscle function in the masseter muscle of miniature swine. American Journal of Physical Anthropology 112 595 613 doi:10.1002/1096-8644(200008)112:4<595::AID-AJPA11>3.0.CO;2-W

Arguello A Lopez-Fernandez JL Rivero JL 2001 Limb myosin heavy chain isoproteins and muscle fibre types in the adult goat (Capra hircus). The Anatomical Record 264 284 293 doi:10.1002/ar.1165

Dennington S Baldwin J 1988 Biochemical correlates of energy-metabolism in muscles used to power hopping by kangaroos. Australian Journal of Zoology 36 229 240 doi:10.1071/ZO9880229

Duris MP Picard B Geay Y 2000 Specificity of different anti-myosin heavy chain antibodies in bovine muscle. Meat Science 55 67 78 doi:10.1016/S0309-1740(99)00127-8

Ford GL Fogerty AC 1982 The fatty acids of kangaroo and wallaby meat. CSIRO Food Research Quarterly 42 57 60

Gardner GE Pethick DW Greenwood PL Hegarty RS 2006 The effect of genotype and plane of nutrition on rate of pH decline post mortem and the expression of enzymatic markers of metabolism in lamb carcases. Australian Journal of Agricultural Research 57 661 670

Gardner GE Hopkins DL Greenwood PL Cake MA Boyce MD Pethick DW 2007 Sheep genotype, age and muscle type affect the expression of metabolic enzyme markers. Australian Journal of Experimental Agriculture 47 1180 1189 doi:10.1071/EA07093

Greenwood PL Davis J Gaunt GM Ferrier GR 2006 a Influences on the loin and cellular characteristics of the m. longissimus lumborum of Australian Poll Dorset-sired lambs. Australian Journal of Agricultural Research 57 1 12 doi:10.1071/AR04316

Greenwood PL Gardner GE Hegarty RS 2006 b Lamb myofibre characteristics are influenced by sire estimated breeding values and pastoral nutritional system. Australian Journal of Agricultural Research 57 627 639 doi:10.1071/AR04318

Greenwood PL Harden S Hopkins DL 2007 Myofibre characteristics of ovine longissimus and semitendinosus muscles are influenced by sire breed, gender, rearing type, age, and carcass weight. Australian Journal of Experimental Agriculture 47 1137 1146 doi:10.1071/EA06324

Greenwood PL Tomkins NW Hunter RA Allingham PG Harden S Harper GS 2009 Bovine myofiber characteristics are influenced by postweaning nutrition. Journal of Animal Science 87 3114 3123 doi:10.2527/jas.2009-1936

Guth L Samaha FJ 1970 Procedure for the histochemical demonstration of actomyosin ATPase. Experimental Neurology 28 365 367 doi:10.1016/0014-4886(70)90244-X

Hopwood PR 1988 Kangaroos as game meat animals: Carcass meat yields and meat inspection. Australian Journal of Zoology 24 169 177

Lucas CA Kang LHD Hoh JFY 2000 Monospecific antibodies against three mammalian fast limb myosin heavy chains. Biochemical and Biophysical Research Communications 272 303 308

Maccatrozzo L Patruno M Toniolo L Reggiani C Mascarello F 2004 Myosin heavy chain 2B isoform is expressed in specialized eye muscles but not in trunk and limb muscles of cattle. European Journal of Histochemistry 48 357 366

Novikoff AB Shin WY Drucker J 1961 Mitochondrial location of oxidative enzymes: Staining results with two tetrazolium salts. The Journal of Biophysical and Biochemical Cytology 9 47 61

O’Dea K 1988 Kangaroo meat – polyunsaturated and low in fat: Ideal for cholesterol-lowering diets. Australian Zoologist 24 140 143

Ouwerkerk D Maguire AJ McMillen L Klieve AV 2009 Hydrogen utilising bacteria from the forestomach of eastern grey (Macropus giganteus) and red (Macropus rufus) kangaroos. Animal Production Science 49 1043 1051

Padykula HA Herman E 1955 The specificity of the histochemical method for adenosine triphosphatase. The Journal of Histochemistry and Cytochemistry 3 170 195

Pette D Staron RS 2000 Myosin isoforms, muscle fiber types, and transitions. Microscopy Research and Technique 50 500 509

Picard B Cassar-Malek I 2009 Evidence for expression of IIb myosin heavy chain isoforms in some skeletal muscles of Blonde d’Aquitaine bulls. Meat Science 82 30 36 doi:10.1016/j.meatsci.2008.11.022

Picard B Duris MP Jurie C 1998 Classification of bovine muscle fibres by different histochemical techniques. The Histochemical Journal 30 473 477 doi:10.1023/A:1003207922944

Reggiani C , Mascarello F (2004) Fibre type identification and functional characterization in adult livestock animals. In ‘Muscle development of livestock animals. Physiology, genetics and meat quality’. (Eds MFW te Pas, ME Everts, HP Haagsman) pp. 39–68. (CAB International: Wallingford, UK)

Rehfeldt C Fiedler I Dietl G Ender K 2000 Myogenesis and postnatal muscle cell growth as influenced by selection. Livestock Production Science 66 177 188 doi:10.1016/S0301-6226(00)00225-6

Sinclair AJ O’Dea K Dunstan G Ireland PD Niall M 1987 Effects on plasma lipids and fatty acid composition of very low fat diets enriched with fish or kangaroo meat. Lipids 22 523 529 doi:10.1007/BF02540369

Spiegel NB (2008) Factors influencing the quality of meat from kangaroos. PhD Thesis, University of Sydney, Australia.

Spiegel NB Wynn PC Thompson JM Greenwood PL 2010 Eastern grey kangaroo (Macropus giganteus) myofibres. 2. Characteristics of eight skeletal muscles. Animal Production Science 50 393 399 doi:10.1071/AN09196

Suzuki A Cassens RG 1983 A histochemical study of myofibre types in the serratus ventralis thoracis muscle of sheep during growth. Journal of Animal Science 56 1447 1458

Tribe DE Peel L 1963 Body composition of the kangaroo (Macropus sp.). Australian Journal of Zoology 11 273 289

Warner RD Pethick DW Greenwood PL Ponnampalam EN Banks RG Hopkins DL 2007 Unravelling the complex interactions between genetics, animal age and nutrition as they impact on tissue deposition, muscle characteristics and quality of Australian sheep meat. Australian Journal of Experimental Agriculture 47 1229 1238 doi:10.1071/EA07229

Wattenburg LW Leong JL 1960 Effects of coenzyme Q10 and menadione on succinic dehydrogenase activity as measured by tetrazolium salt. The Journal of Histochemistry and Cytochemistry 8 296 303

Wilson GR Edwards MJ 2008 Native wildlife on rangelands to minimise methane and produce lower-emission meat: Kangaroos versus livestock. Conservation Letters 1 119 128

Zhong WWH Lucas CA Kang LHD Hoh JFY 2001 Electrophoretic and immunochemical evidence showing that marsupial limb muscles express the same fast and slow myosin heavy chains as eutherians. Electrophoresis 22 1016 1020 doi:10.1002/1522-2683()22:6<1016::AID-ELPS1016>3.0.CO;2-K

Zhong WWH Lucas CA Hoh JFY 2008 Myosin isoforms and fibre types in limb muscles of Australian marsupials: adaptations to hopping and non-hopping locomotion. Journal of Comparative Physiology B, Biochemical, Systemic, and Environmental Physiology 178 47 55 doi:10.1007/s00360-007-0198-8

Export Citation Cited By (2)