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Serum LDH isoenzyme activity in dairy and beef cows

Article  in  The Indian veterinary journal · July 2004

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 Medicina Veterinaria 1(2) 2002, 39-43

SERUM LDH ISOENZYME ACTIVITY

IN DAIRY AND BEEF COWS

Przemysław Sobiech, Zygmunt Kuleta, Marek Jałyński

Abstract. The total serum LDH activity was determined in beef and dairy cows. LDH
was also divided into isoenzymatic fractions. The total activity of lactate dehydrogenase
was the highest in Limousine cows, and the lowest – in H-F ones. 5 LDH isoenzymes
were present in all of the breeds examined. The activity of fractions LDH1 and LDH2 was
higher in dairy cows, whereas the activity of fractions LDH3, LDH4, and especially LDH5
– in beef ones.

Key words : cows, LDH, isoenzymes

INTRODUCTION

The development of diagnostic methods allows to diversify laboratory determina-

tions. One of the indices determined more and more frequently in the diagnostics of
metabolic diseases is the total activity of lactate dehydrogenase and its isoenzymes.
Isoenzymes are various molecular forms of the same enzyme, catalyzing the same
chemical reaction and differing from one another in tissue affinity, enzymatic kinetics,
electrophoretic mobility, chromatographic and immunological properties [Hamm 1990,
Murray et al. 1994]. Enzyme heterogeneity is a very common phenomenon, showing
how tissues and organs adapt to certain environmental conditions. It is used in labora-
tory diagnostics, as damaged tissues and organs release to circulating blood this molecu-
lar form of a given enzyme in which they abound.
From the clinical perspective, the determination of serum isoenzyme activity is of
primary importance, but its determination in biological material from various tissues
and organs is also useful.
The most observations concerning enzyme heterogeneity were made in relation to
lactate dehydrogenase. This enzyme is present in all cells where the process of glycoly-
sis takes place. Five different LDH forms, referred to as fractions LDH1, LDH2, LDH3,
LDH4 and LDH5, can be found in biological fluids and tissue homogenates. This divi-
sion is based on the speed of their movement in an electric field [Hamm 1990] – the
fraction LDH1 moves at the highest rate.
A single LDH molecule is a tetramer built of two subunits – subunit H (heart) and
subunit M (muscle). The fraction LDH1 consists of subunits H only, whereas the frac-
tion LDH5 contains only subunits M. Isoenzymes characterized by moderate speed of
40 P. Sobiech et. al.

movement in an electric field are mixed forms and contain different numbers of sub-
units H and M. Both types of subunits are polypeptide chains with molecular weight of
ca 35 000 and similar antigen properties [Angielski 1985]. LDH catalyzes a reversible
reaction of pyruvate conversion into lactate in the presence of NADH. Differences in
the tissue activity of particular isoenzymes result from the fact that cathodic fractions
(LDH4 and LDH5) catalyze this reaction when considerable amounts of lactate are ac-
cumulated (i.e. when anaerobic metabolism dominates), whereas anodic isoenzymes
(LDH1, LDH2 and LDH3) can be found in tissues with aerobic pyruvate conversion in
the tricarboxylic acid cycle [Wang et al. 1997].
In cattle the highest total activity of LDH was observed in the cardiac muscle, liver
and kidneys. Fractions LDH1 and LDH2 dominate in these organs – ca. 35% of the total
enzyme activity. Anodic fractions dominate also in bovine serum, LDH1 – 40% of the
total enzyme activity, LDH2 – 25% [Keller 1974, Yasuda et al. 1989].
First reports on the possibility to use LDH isoenzymatic separation in disease diag-
nosis in ruminants appeared in the sixties, when Boyd [1964] noted a significant in-
crease in the activity of LDH5 in lambs with acute muscular distrophy.
In cattle an increase in the activity of LDH1 was observed during experimental liver
damage, whereas the activity of LDH5 increased in the case of experimental skeletal
muscle damage. The activity of LDH isoenzymes in bovine serum was also examined in
the course of various neoplastic processes. It was found that in such cases the activity of
fractions LDH1 and LDH2 increases, similarly as in liver tissue degeneration [Yasuda et
al. 1989].
The aim of the studies was to determine and compare serum LDH isoenzymatic pro-
files in the most common breeds of dairy and beef cattle in Poland.

MATERIAL AND METHODS

The studies were conducted on 50 cows, 10 from each of the following breeds:
NCB, Jersey, H-F (dairy breeds), Aberdeen Angus and Limousine (beef breeds).
NCB cows were four to six years old, with body weight of ca. 500 kg. Their milk
productivity was at an average level of 5 000–5 500 liters per year. Their daily ration
consisted of 30 kg of silage, 4 kg of hay and 4 kg of mixture B. They were given water
ad libitum.
Jersey cows were four years of age, with body weight of ca. 400 kg. The milk yield
was in their case ca. 5 000 liters per year. Their daily ration included 25 kg of silage,
4 kg of mixture B, 4 kg of hay, beets and molasses, supplemented with minerals and
vitamins.
H-F cows were aged four to five years, with body weight of ca. 600 kg and an aver-
age milk yield at a level of 8 800 liters per year. They received each day 30 kg of silage,
4 kg of soybean-maize concentrate (Central–Soya Olsztynek), 4 kg of hay and a premix
for dairy cattle.
Aberdeen-Angus cows were four to five years old, with body weight of 600–650 kg.
Their daily ration consisted of 30 kg of feed concentrate, 6 kg of hay, 5 kg of soybean-
-maize concentrate and mineral supplements.
Limousine cows were four to six years of age, with body weight of 650 kg. Their
daily ration was similar to that of Aberdeen-Angus cows.

Acta Sci. Pol.

Serum LDH isoenzyme ... 41

Their blood was collected for analyses from the jugular vein, in the morning, imme-
diately after the first milking of dairy cows. All animals were subjected to clinical and
parasitologic examinations. No diseases or internal parasites were found. Laboratory
analyses included the determination of serum LDH activity by the enzymatic method
(Kit Analco, 300C), and LDH isoenzyme activity by high-voltage electrophoresis on
agarose (system Paragon, Beckman).
The results were analyzed statistically by the Student’s t-test.

RESULTS AND DISCUSSION

The total activity of lactate dehydrogenase was different in particular cattle breeds.
Its average level was the highest in Limousine cows – ca. 2 500 U/l, and the lowest – in
H-F ones – ca. 1 740 U/l (Table 1). However, it should be emphasized that the differ-
ences noted between the breeds examined remained within reference values for this
species.

Table 1. Total average serum activity of LDH and its isoenzymes in cows of various breeds
(x ± S)
Tabela 1. Całkowita średnia aktywność LDH i aktywność jej izoenzymów w surowicy krów
róŜnych ras (x ± S)
Aberdeen Holstein Fri-
NCB Limousine Jersey
Angus ziens
LDH U/l 1822.5 ±146.4 2505.5 ± 90.9 1883.8 ± 151.4 2278.1 ± 190.1 1743.5 ± 283.8
LDH1 % 50.9 ± 2.9 37.3 ± 5.4 42.5 ± 2.7 51.1 ± 2.8 52.5 ± 3.5
LDH2 % 27.1 ± 1.3 30.9 ± 2.1 30.3 ± 1.1 30.1 ± 1.1 28.1 ± 1.4
LDH3 % 14.3 ± 1.4 21.5 ± 3.8 18.1 ± 0.9 13.3 ± 1.4 13.4 ± 1.8
LDH4 % 4.3 ± 0.9 5.6 ± 1.1 5.6 ± 0.9 3.4 ± 0.5 3.5 ± 0.5
LDH5 % 3.5 ± 0.9 a 4.5 ± 2.4 3.5 ± 0.7 a 2.1 ± 0.5 a 2.6 ± 1.1

a – statistically significant difference for p ≤ 0.05

a – róŜnica statystycznie istotna dla p ≤ 0.05

Serum isoenzymatic separation showed that five LDH fractions were present in all
of the cows. The activity of LDH1 in beef cows was statistically significantly lower than
in dairy ones. This could be connected with higher productivity of dairy cows, i.e. a
higher liver load, as in ruminants isoenzyme LDH1 is present in the cardiac muscle and
liver, and changes in its activity reflect the functional condition of these organs. Similar
results were obtained by Ashmare et al. [Asefa et al. 1998] who studied LDH isoen-
zyme activity in dairy cows at different stages of lactation. They observed a positive
correlation between LDH1 activity and organism productivity.
Similarly to the activity of LDH1, the activity of LDH2 was higher in the serum of
dairy cows, whereas the activity of isoenzymes LDH3, LDH4 and LDH5 (especially of
the last two ones) was considerably higher in beef cows (Table 1). This is natural, as in
ruminants LDH4 and LDH5 are characteristic of skeletal muscles, and muscle weight is
very high in beef cows.

Medicina Veterinaria 1(2) 2002

42 P. Sobiech et. al.

CONCLUSIONS

The activity of all five LDH fractions, including LDH5, was determined in the pre-
sent studies in all of the experimental cows. This is worth noting as some researchers
found it difficult to isolate this fractions [Asefa et al. 1998, Asefa et al. 1999, Tollersrud
1970], both in healthy cows and in those with metabolic diseases. The absence of this
fraction has not been fully explained. According to some authors, it could be connected
with its thermolability fractions [Asefa et al. 1998, Tollersrud 1970]. In the light of our
investigations it seems that the reason could be equipment imperfection or improper
methodology. The determination of serum LDH isoenzyme activity by the system Para-
gon (Beckman) enabled full isoenzymatic separation of LDH and guaranteed high reli-
ability of results. An analysis of the results obtained indicates also that particular cattle
breeds show no predisposition to the presence or absence of certain LDH isoenzymatic
fractions in their serum, as suggested by some authors [Asefa et al. 1998, Asefa et al.
1999]. To sum up, it may be stated that the activity of LDH1 and LDH2 is higher in
dairy cows, whereas the activity of LDH4 and LDH5 is higher in beef cows. This fact
should be kept in mind while interpreting LDH separation in cattle in the diagnostics of
heart, liver and skeletal muscle diseases.

REFERENCES

Angielski S. (red), 1985. Biochemia kliniczna i analityka.Wyd.2., PZWL, Warszawa.

Asefa A., Kovac G., Reichel P., Buleca J., Scurokova E., 1998. Serum isoenzyme activity of
lactate dehydrogenase in dairy cows at different stages of milk production. Folia Veterinaria,
42, 2: 77–81.
Asefa A., Kovac G., Reichel P., Scurokova E., 1999. The behaviour of serum LDH isoenzyme
activity in patients toward injected antibiotics and breeds of dairy cows. Folia Veterinaria, 43,
2 : 61–66.
Boyd J.W., 1964. Serum enzyme changes in lambs with experimentally-induced acute muscular
dystrophy. Res. Vet. Sci., 5 : 419–433.
Hamm R., 1990. Lactate dehydrogenase and its importance in meat research. Physiological func-
tion of isozymes; LDH structure; distribution and binding of LDH in the muscle cells.
Fleischwirtschaft., 70, 11 : 1336–1339.
Keller P., 1974. Lactate dehydrogenase isoenzymes in normal bovine serum and during experi-
mental liver and muscle damage. Res. Vet. Sci., 17, 1 : 49–58.
Murray R.K., Granner D.K., Mayes P.A., Rodwell V.W., 1994. Biochemia Harpera. Wyd. 3.,
Wydawnictwo Lekarskie PZWL, Warszawa.
Tollersrud S., 1970. Heat stability of serum lactate dehydrogenase and its isoenzymes in young
and adult cattle and sheep. Acta Vet. Scand., 11, 4 : 510–524.
Wang X.C., Jiang L., Zhou H.M., 1997. Minimal functional unit of lactate dehydrogenase. J.
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Acta Sci. Pol.

 Serum LDH isoenzyme ... 43

AKTYWNOŚĆ IZOENZYMÓW LDH W SUROWICY KRÓW

RAS MLECZNYCH I MIĘSNYCH

Streszczenie. W surowicy krów ras mięsnych i mlecznych wykonano oznaczenia całko-

witej aktywności LDH i dokonano rozdziału na jej frakcje izoenzymatyczne. Całkowita
aktywność dehydrogenazy mleczanowej była największa u krów rasy Limousine, a naj-
mniejsza u krów rasy H-F.
U wszystkich ras wykazano obecność 5 izoenzymów LDH, przy czym aktywność frakcji
LDH1 i LDH2 była większa u krów ras mlecznych, a aktywność frakcji LDH3, LDH4,
a szczególnie LDH5 była zdecydowanie wyŜsza u krów ras mięsnych.

Słowa kluczowe: krowy , LDH, izoenzymy

Przemysław Sobiech, Zygmunt Kuleta, Department of Internal Diseases, Marek Jałyński,

Department of Surgery and Rentgenology, University of Warmia and Mazury, Olsztyn, Poland.
psobiech@moskit.uwm.edu.pl, ul. Oczapowskiego 14, 10–957 Olsztyn.

Medicina Veterinaria 1(2) 2002

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