LECTURE - FRAUDULENT SUBSTITUTION OF MEAT AND ITS RECOGNITION

By Dr. Subhash S, Raut

 Replacement of conventional meat by similar looking inferior quality or non-

conventional sources is known as fraudulent substitution of meat or falsification of
meat.
 It may include
- Mixing meat of different animals
- Mixing or replacement of inferior/undesirable meat with superior/costly meat.
E.g. beef with horse flesh
Mutton with chevon
Rabbit meat with cat flesh etc.
 The methods classified as follows:
o Physical methods
 This is based on general appearance, colour, texture, odour and
tenderness of different species of meat.
 Besides the general characteristics of body fat, its colour, marbling,
firmness of fat can be identified.
 Dentition formula, vertebral formula and articulation pattern, rib number
and degree of curvature, characteristics of long bones will also help to
identify the species if the carcass is intact.
o Chemical Methods
 Linoleic acid content
 Iodine value
 Refractive index of fat
 Myoglobin content
o Biological Methods
 Electrophoretic method
 Immunological/serological method
 Latest techniques
PHYSICAL METHODS
 Physical methods like anatomical differences of each species of the carcass and
appearance of muscle and fat colour, odour, texture and taste have provided a general
difference between species in earlier days for food analysis.
 So, this can be attempted, provided the meats are in the form of joints and in carcass
form.
Carcasses of different species of food animals
 Horse
 o Neck and the bones of limbs are longer than the ox.
o Sternum of horse is canoe shaped.
o No diarthrodial joint between the first and second sternal ribs.
o There are 18 pairs of ribs and are narrower than those of ox.
 Bull
o The outstanding characteristic in the bull carcass is the massive development of
the muscles of the neck and shoulder and also in the hindquarters of the well-
bred animals.
o Neck is much thicker than that of the ox.
o Ligamentum nuchae is thicker and stronger than in ox.
o Anterior part of the ischio pubic symphysis is well developed and forms a
distinct tubercle.
o Inguinal canals are patent.

 Ox
o Shows lesser muscular development than that of bull especially in the neck and
shoulder region.
o There is even covering of fat on the exterior.
o The scrotal fat is prominent, nodular and more or less pointed. Pelvis is narrow
and usually contains a relatively large quantity of fat.
o Fat is usually plentiful over the kidneys and along the sublumbar region.
 Cow
o Thigh is less rounded than that of ox.
o This is very noticeable in the hind quarters (sunken round).
o The pelvis is broader. Anterior tubercular pelvis is broader.
o Udder is present, if removed triangular area of attachment is noticeable on each
side of midline of the abdominal wall.
o In heifers the udder is only slightly developed and consists chiefly of fat.
o In old cows the udder is soft, spongy, round and pendulous.
 Sheep
o The carcass of sheep (whether or ewe) is characterised by an abundant and even
distribution of subcutaneous fat.
o The carcass of ram is distinguished by great muscular development in the region
of neck and shoulders; the ligamentum nuchae is large and strong.
o The neck is thick and the inguinal canals are patent.
 Goat
o Goats are long and lean.
 o There is very little subcutaneous fat, kidney fat abundant even in poor carcasses.
o Subcutaneous connective tissue is sticky in nature and during skinning loose
hairs from the skin become adherent to the subcutaneous tissue and cannot be
removed completely.
o Pelvis of goat is long and narrow.
 Hog
o Carcass of pig cannot easily be mistaken for that of any other animal.
o In most countries the skin is left on the carcass.
o But even when the skin is removed there should be no difficulty in
identification.
DIFFERENTIATION OF CARCASSES OF ANIMALS
Differentiation of carcasses of horse and ox
 Carcass of the horse and ox may be differentiated by the following details
o In the horse the unusual length of the sides is noticeable, together with the great
muscular development of the hindquarters.
o The thoracic cavity is longer in the horse; this animal possesses 18 pairs of ribs,
whereas the ox has 13 pairs.
o The ribs in the horse are narrower but more markedly curved.
o The superior spinous processes of the first six dorsal vertebrae are more
markedly developed in the horse and are less inclined posterior.
o In the forequarter, the ulna of horse extends only half the length of the radius;
in the ox it is extended and articulates with the carpus.
o In the hindquarter, the femur of the ox possesses no third trochanter; the fibula
is only a small pointed projection, but in the horse it extends two–third the
length of the tibia.
o In the horse the last three lumbar transverse processes articulate with each other,
the sixth articulating in a similar manner with the sacrum.
o They do not articulate in the ox.
o The horse carcass shows considerable development of soft yellow fat beneath
the peritoneum, especially in the gelding and mare, but in the stallion the fat is
generally of a lighter colour and almost white. In the ox the kidney fat is always
firmer, whiter and more abundant than in the horse.
o Horseflesh is a dark red, initially brown or reddish brown on exposure to
atmosphere the colour turns bluish.
o Marbling is absent in horsemeat; it is firm but sticky in nature due to high
glycogen content.
o Horsemeat has a pronounced sweet taste, repulsive odour and well defined
muscle fibre.
o Beef lack the bluish tinge.
Differentiation of carcasses of sheep and goat
 Features Sheep Goat

Back and withers Round and well fleshed Sharp, little fleshed

Thorax Barrel shaped Flattened laterally

Tail Fairly broad Thin

Radius 1.25 times length of Twice as long as metacarpus

metacarpus

Scapula Short an broad, superior Possess distinct neck. Spine straight and narrow,
spine, bent back and lateral border thin and sharp
thickened

Sacrum Lateral borders thickened in Sharp

the form of rolls

Flesh Pale red and fine in texture Dark red and coarse with goaty odour. Sticky
subcutaneous tissue, which may have adherent
goat hairs.

Sheep, Goat and Deer

 Among these carcasses, in deer, the scapula’s acromion is elongated into a sharp point,
which is directed ventrally.
 The acromion is absent in the sheep and goat or is considerably smaller.
 The radio-ulna arch, which forms an oval opening in the sheep and goat, is very long
in deer.
 In deer, the subcutaneous layer of fat is not as well developed as in sheep.
 The meat is poor in fat and possesses the odour of venison, which is easily
distinguishable from the odour of sheep.
Hog and Dog
 The colour of dog meat is very darker than pork and is easily made out in cooked meat.
 The muscles of the dog are searier and the fat is oilier than hog fat.
 The odour of the dog meat is repulsive.
Cat and Rabbit
 The meat of the cat is paler than rabbit meat.
 The fat of the cat appears whitish in contrast to rabbit fat, which is honey yellow.
Meat and Fat of Sheep and Dog
 The meat and fat of sheep and dog are indistinguishable by the naked eye and the
carcasses of large dogs are sometimes substituted for mutton.
 The ribs and sternum of the sheep are broad and flat, while those of the dogs are round
in section.
  In the hind leg, the sheep has only one bone, the tibia articulating with the tarsal joint,
while the dog has both tibia and fibula.
 The sheep has triangular scapula with a broad, prolonging cartilage and the radius and
ulna lies close together for their whole length, while the scapula of the dog has a semi-
circular posterior upper edge with practically no prolonging cartilage and the radius and
ulna are widely separated along the greater part of their shafts.
 The xiphoid cartilage in sheep is firm and grisly, while in the dog, it is softer and florous
and shaped like a dagger.
Cattle and Buffalo
 Generally fresh buffalo meat is darker (more reddish brown) and the fibres are coarser
and looser in structure than beef.
 The odour of the buffalo meat and fat are always strikingly musky and if boiled in
strong acidified (H2SO4) water, it develops a disagreeable odour similar to that of cattle
manure.
 The cutaneous shoulder muscle of buffalo is only 3 to 4 finger broad, while that of cattle
it is considerably broader.
 The fat of buffalo is strikingly white and drier and less sticky than in cattle.
 The confirmation of the bones of the buffalo is generally thinner and the bones are very
brittle.
 The ischio pubic symphysis of the buffalo is strikingly plane.
CHARACTERISTICS OF MEAT
Horse meat
 The meat of horse is dark red in colour, on exposure to air acquires a bluish tinge or
shield on the surface and later become very dark.
 Odour is peculiar – sweet and to most people more or less repulsive. Horseflesh
contains large quantities of glycogen (2%).
 Fat is yellow or brownish yellow in colour and owing to its high olein content it is soft
and greasy.
Beef
 The colour of beef varies from light red to dark red according to the age and the part of
the carcass from which it was collected.
 The meat is moist, silky to the touch and is marbled with fat.
 Fat is fine usually creamy white or yellowish white in colour.
 In old cattle the fat tends to be more yellow and somewhat loosen in consistency.
 In Jersey and Guernsey fat is pronounced yellow colour.
 Meat of heifer closely resembles that of young ox.
 The meat of old cow is not marbled and tends to be lean, dry and somewhat coarse.
 However, the meat and fat of old dairy cows are often relatively moist.
 The veal is pale or grayish red in colour.
  Not very firm under pressure of fingers. Fibres are tough.
Mutton
 The meat of wether or ewe varies in colour from light red through brownish red to dark
red.
 According to the age of the animal and to the part of the carcass – the fibres are fine,
dense and firm. Marbling with fat is practically absent.
 The fat is white, very firm and odourless.
Goat meat/Chevon
 Chevon is not marbled and bears a fairly close resemblance to that of sheep.
 The meat of uncastrated adult goat has a goaty odour.
Pig
 The meat of pig varies in colour according to the age and nutritive condition of the
animal and also according to the body region from which it is derived.
 It may be pale red, reddish gray; rose red, dark red or in certain parts may be almost
colourless.
 It is less firm to the touch than other food animals.
 The fibres are fine, fat is white, soft and greasy.
CHARACTERISTICS OF FAT

S. No Species Colour Consistency

1. Cow Yellow Fairly firm

2. Bull; heifer Yellowish white Firm

3. Calf White or grayish white Soft and gelatinous

4. Buffalo Strikingly white Fairly firm

5. Sheep ; Goat Very white Typically crispy in sheep. Very firm.

6. Pig Generously white Fairly firm, greasy and not crispy

7. Horse Yellowish white Soft and greasy

CHEMICAL METHODS
 The chemical tests consist of the determination of
o the content of glycogen in flesh
o the percentage of linoleic acid in fat
o the amount of iodine absorbed by unsaturated fatty acids in fat and
 o the refractive index of fat.
Test for Glycogen Content of Meat
 The horseflesh is richer than the flesh of other food animals in glycogen in horsemeat
as compared with other kinds of meat, glycogen is found in large quantities irrespective
of age.
o Horse – 0.5 to 1.0 %
o Beef - 0.0 to 0.5%
o Pork and mutton - nil
 Disadvantages
o The flesh should be tested for the content of glycogen soon after the slaughter
as it disappears from the flesh quickly.
o Liver of all food animals especially pig liver contains more glycogen when they
are used in sausage making it gives a high percentage. So care must be taken in
interpretation of results.
Linoleic acid content
 Horse fat contains 1-2% linoleic acid. Linoleic acid content in other animals' fat is not
more than 0.1%.
 Thus adulteration of lard or beef and mutton fat with horse fat can be identified by
estimation of the linoleic acid concentration.
Iodine value
 Estimation of iodine value is a valuable test for the detection of horse fat.
 Iodine value is the amount of iodine absorbed by the unsaturated fatty acid present in
the fat.
 Good lard has an iodine value of 66.
 The iodine value of the fat from various food animals is:
o Horse - 71-86
o Ox (cattle) - 38-46
o Sheep - 35-46
o Pig - 50-70
Refractive index
 Refractive index is another valuable test for the detection of fat of different animal
species.
 Fat is liquefied by heat and converted into oil for estimation of refractive index.
 All liquids including oils possess a specific refractive index.
o Horse - 53.5
o Ox - less than 40
o Pig - not above 51.9
Myoglobin content
 The myoglobin content of different species is:
 o Beef - 0.30 to 1%
o Pork - 0.06 to 0.40%
o Poultry - 0.02 to 0.18%
Other chemical tests for differentiation of different meat species are:
 Myoglobin content
 Muscle enzymes
 Composition of fat
 Carotene content
 Fatty acid composition
ELECTROPHORETIC METHODS
 The various electrophoretic methods include
o Agar gel electrophoresis
o Polyacrylamide Disc electrophoresis
o Polyacrylamide gel electrophoresis
o Sodium dodecyl sulfate polyacrylamide gel electrophoresis
o Iso-electric focussing
 Agar gel electrophoresis
o Electrophoretic separation of proteins on the basis of mobility of proteins in a
supporting gel of agarose at a constant pH and electrical field is termed as Agar
gel electrophoresis.
 Polyacrylamide Disc electrophoresis
 Polyacrylamide gel electrophoresis (PAGE)
o Polyacrylamide gels are prepared using acrylamide which provides cross
linking between the polymerized long chains of acrylamide.
o The separation of proteins on the basis of their mobility in a polyacrylamide gel
is comparatively better since the resolution of different proteins are optimum.
Polyacrylamide gel electrophoresis also requires a constant pH and electrical
field to provide high resolution of protein.
 Iso-electric focussing (IEF)
o It is an electrophoretic technique which utilizes the charge at the surface of the
protein to drive it through a gradient gel.
o The pH gradient is setup by polyacrylamide compounds, the ampholytes.
o The proteins applied on to the gel reach a point where the surface charges
becomes neutral at their iso electric point
IMMUNOLOGICAL METHODS
 Immunological methods include
o Precipitation test
 o Complement fixation test (CFT)
o Double immuno diffusion test (AGID)
o Single radial immunodiffusion test
o Enzyme linked immuno sorbent assay (ELISA)
Double Immuno Diffusion Test (AGID)
 Based on a simple double diffusion method (Ouchterlony, 1948). Species specific
antiserum (antibody) and unidentified meat extract (antigen) are allowed to diffuse
towards one another in an agar gel slab. If the antigen and antibody are homologous a
precipitin band is formed along the line where the two meet.
Single Radial immunodiffusion test
 In this test serum albumen content in meat extract is estimated that only this protein
formed an immune precipitate in antiserum.
 Immunological species specificity of albumen is regarded as far superior than the
globulin formation.
Enzyme Linked Immuno Sorbent Assay (ELISA)
 The technique involves the application of species specific antibodies to the proteins
(antigen) coated on the plastic surface of micro-titre plate.
 The recognition of antigen results in the formation of antigen-antibody complex, which
are detected by either enzyme linked immunoglobulin or protein A (antibody detector)
producing visible colour reaction with added substrate.
 The colour intensity is measured objectively at a specific wave length in a micro ELISA
reader as absorbance value.
Recent techniques in meat species identification include
 Production of monoclonal antibodies and application of ELISA.
 Use of DNA probes and application of DNA hybridization technique.
 Use of Polymerase Chain Reaction (PCR) in species identification.
RECENT TECHNIQUES IN MEAT SPECIES IDENTIFICATION
 Recent techniques in meat species identification include
o Production of monoclonal antibodies and application of ELISA.
o Use of DNA probes and application of DNA hybridization technique.
o Use of Polymerase Chain Reaction (PCR) in species identification