VETERINARSKI ARHIV 68 (3), 143-148, 1998

ISSN 0372-5480
Printed in Croatia

The influence of the addition of enzymes in feed on histochemical changes in white and dark chicken meat

Ante Jelic*, Srebrenka Nejedli, Ante Hraste, Mladen Zobundzija, and Karmen Botka Petrak

Department of Anatomy, Histology and Embryology, Faculty of Veterinary Medicine,
University of Zagreb, Zagreb, Croatia

* Contact address:
Dr. Ante Jelic,
Department of Anatomy, Histology and Embriology, Faculty of Veterinary Medicine, 10000 Zagreb, Heinzelova 55, Croatia,
Phone: 385 1 23 90 242, Fax: 385 1 214 697

JELIC, A., S. NEJEDLI, A. HRASTE, M. ZOBUNDZIJA, K. BOTKA PETRAK: The influence of the addition of enzymes in feed on histochemical changes in white and dark chicken meat. Vet. arhiv 68, 143-148, 1998.

ABSTRACT

In the study, Jata-line chickens were used. For the experiment twenty chickens were equally divided into two groups - a control group (C) and an experimental group (E). During the whole 42-day period of fattening the chickens were fed with a pasture mixture in which maize was the main component. During the period of fattening the experimental group received feed mixed with the polyenzymatic preparation in a ratio of 0.5/1000 kg. The addition of polyenzymatic preparation to chicken feed resulted in a larger diameter of the fibre in m. pectoralis superficialis and m. iliofibularis, and in the percentage of white quick-contractible glycolytic fibre in m. iliofibularis in chickens.

Key words: enzymes, chicken, histology, m. iliofibularis, m. pectoralis superficialis, chicken meat

Introduction

The first results of enzymes were applied in poultry feed and described in the 1950s (FRY et al., 1958). It is considered that enzymes in polyenzymatic preparation have a synergetic effect (NAESI, 1988) and give better results compared to the individual addition of enzymes (INBORR, 1990). Accordingly, in our research on Jata-line chickens during fattening, we mixed corn with a powder polyenzymatic preparation protease, where corn was the dominating substance in the feed. We focussed our research on the muscles, as the aim of breeding these animals is meat production.

Materials and methods

In our test we used one-day-old male chickens from the leading line "Jata". The chickens were divided in two groups: Control (C) and experimental (E) with ten birds in each group. The chickens were kept in a deep litter of sawdust during the whole fattening period.

During 42 weeks of fattening the broilers of both groups were fed with a feed mixture containing a majority of corn (61.4%). In the feed of group (E) we mixed the preparation which contains the enzymes alfa-amilase, beta-glukonase, beta-glukosidase, celulase and protease, to a quantity of 0.5/1000 kg. During fattening we monitored conversion and feed digestion.

At the end of the fattening period the chickens were slaughtered.

The tests were made on m. pectoralis superficialis and m. iliofibularis. Pieces of tissue were taken from the right m. pectoralis superficialis in the regio thoracalis and m. iliofibularis of the right leg.

For histochemical research the samples were frozen in fluid nitrogen, cut to 10 µm-thick slices. These were treated through a procedure for review of the activity of lactic acid dehydrogenase (HESS et al., 1958) and succinic acid dehydrogenase (PADYKULA, 1952) acid and alkalicstabile myosin adenosin 3-phosphatase (PADYKULA and HERMAN, 1955), and for a review of lipid content by Sudan Black B treatment according to Lison (ROMEIS, 1968) and for the review of connective tissue, according to Van Gieson (ROMEIS, 1968).

In addition to histochemical research we used the measuring treatment of histological photographs, with a review of lactic acid dehydrogenase. Measurement was made by Leitz microscopic scale, taken under the same enlargement (10 × 6.3) as the histological preparations. We also measured the diameter of muscle fibre.

Results

In the m. pectoralis superficialis of chickens in groups C and E, only white quick-contractible glycolytic fibres were present. This showed a weak activity of lactic and succinic acid dehydrogenase and sour stabile miosin adenosin-3-phosphatase, and a strong alkalic stabile miosin adenosin-3-phosphatase activity.

In chickens of groups C and E in fibres of m. pectoralis superficialis there was no presence of lipids observed. Lipid drops of various shapes were observed only in connective tissue between muscle fibres. Inside the diagonal diameter of m. pectoralis superficialis in chickens of groups C and E, fibres with distinct permisial connective tissue strips and tender endomisyum which surrounded each muscle fibre.

In m. pectoralis superficialis the median value of fibre diameter in chickens of group E was 42.05 µm, and in group C was 38.95 µm. This enlargement of fibres was statistically significant (P<0.01). (Table 1). In m. iliofibularis the median fibre diameter in chickens of group E was 36.05 µm and was larger than in group C, where it was 32.00 µm, which is statistically significant (P<0.01). (Table 2).

On the basis of the studied enzyme activity in m. iliofibularis of chickens in groups C and E, two types of fibres were observed: red, which showed a medium to very strong activity of lactic and succinic and dehydrogenase and sour stabile miosin adenosin-3-phosphatase, and a weak activity of alkalic stabile miosin adenosin-3-phosphatase; and white, which showed weak activity of lactic and succinic acid dehydrogenase and sour stabile miosin adenosin-3-phosphatase, and a strong activity of alkalic stabile miosin adenosin-3- phosphatase.

Table 1. Fibre diameters in m. pectoralis superficialis of ten chickens in control (C) and ten chickens in experimental (E) groups (in µm). Experimental group was fed with an addition of 0.5/1000 kg polyenzymatic preparation in feed over a 42-week period.

*P<0.01

Table 2. Fibre diameters in m. iliofibularis in ten chickens of control (C) and ten chickens of experimental (E) groups (in µm). Experimental group was fed with an addition of 0.5/1000 kg polyenzymatic preparation in feed during a 42-week period.

*P<0.01

Results showed that in m. iliofibularis of chickens in group C there was the largest number of red slow-contractible fibre (85%) and a lower number of white quick-contractible fibre (15%) (Fig. 1). In m. iliofibularis of chickens in group E there was the largest number of red fibres (60%) and a lower number of white fibres (40%) (Fig. 2).

The largest number of fibres in m. iliofibularis in chickens in groups C and E showed a larger quantity of lipids and a lower number of fibre, but no lipid presence. Fibres in m. iliofibularis of chickens in groups C and E were surrounded by a larger quantity of perimysium and endomysium connective tissue than in m. pectoralis superficialis.

Fig. 1. M. iliofibularis, control (C) group of chickens, lactic acid dehydrogenase; enlargement 10×6.3; bar = 100 µm.

Fig. 2. M. iliofibularis, experimental (E) group of chickens, lactic acid dehydrogenase. Experimental group was fed with an addition of 0.5/1000 kg enzymes in feed over a 42-week period. Enlargement 10×6.3; bar = 100 µm.

Discussion

Growth in the diameter of muscle fibre leads to the growth of muscle matter, and the variation in the size of fibre diameter depends on the genetic base, method of breeding and feeding of animals (STAUN, 1972).

It is well known that enzymes added to food act in such a way as to improve digestibility of feed, which could have an effect on better growth and in enlargement of muscle fibre diameter (KAUFMAN, 1997). We wanted to examine whether the polyenzymatic preparation, in a ratio of 0.5/1000 kg, is able to function as a bio-stimulator. For this purpose it was interesting to determine how the addition of this preparation to feed effects the diameter of muscle fibre.

Chickens in test group E showed that in m. pectoralis superficialis (7.97%) and m. iliofibularis (12.65%) a larger diameter of muscle fibre was achieved. We are therefore able to conclude that the polyenzymatic preparation added to a ratio of 0.5/1000 kg feed essentially effected the growth of muscle fibre diameter on both researched muscles.

The aim of our research was to prove how the polyenzymatic preparation added to feed (0.5/1000 kg) of chickens undergoing fattening can influence histochemical changes in white (m. pectoralis superficialis) and in dark (m. iliofibularis) muscles.

The obtained results indicate that the m. pectoralis superficialis of chickens in control group C and chickens in test group E consist only of white quick-contractible glycolytic fibre. This is agreed with the findings of COSMOS (1970). In m. iliofibularis there are two types of fibre: white quick-contractible glycolytic fibre, and red slow-contractible fibre. Adding the polyenzymatic preparation to feed for group E chickens, the type and diameter of muscle fibre was altered. Chickens in group E had a larger number of white fibres (25%) compared with red fibres.

The results of our research showed that in m. iliofibularis of chickens in test group E compared to those in control group C, the number of white fibres increased, which probably influenced the enlargement of the whole muscle matter and improved its quality. This indicates that the studied polyenzymatic preparation acted as a bio-stimulator.

References

COSMOS, E. (1970): Quotageny of red and white muscles: the enzymatic profile and lipid distribution of innature and nature muscles of normal and dystrophic chickens. In: The Physiology and Biochemistry of Muscles as a food. (E. J. Briskey, R. G. Cassens, B. B. Marsh, Eds.). The University of Wisconsion Press. Madison, Midealuhe and London.

FRY, R. E. J., B. ALLRED, L. S. JENSEN, J. McGINNIS (1958): Influence of enzyme supplementation and water treatment on the nutritional value of different grains of poults. Poultry Sci. 37, 372-375.

HESS, R., D. G. SCARPELLI, A. E. G. PEARSE (1958): The cytochemical localization of oxidative enzymes: II Pyridine nucleotide-linked dehydrogenase. J. Byophis. Biochem. Cytol. 4, 753-768.

INBORR, J. (1990): Practical application of feed enzymes. Feed Coumpounder. Creative Press. London.

KAUFMAN, S., A. JELIC, A. HRASTE, K. BOTKA PETRAK, D. NEJEDLI (1997): Histochemische Veränderungen in Geflügefleisch. Fleischwirtsch. 77, 479-481.

NAESI, M. (1988): Enzyme supplementation of laying hen diets based on barley and oats. In: Biotechnology in feed industry. Proceedings of Alltech's 4^th Annual Symposium (Lyons T. P., Ed). Alltech tecnical publictions. Nicholasville, Kentucky. pp. 199-204.

PADYKULA, H. A. (1952): The localization of succinic dehydrogenase in tissue sections of the rats. Am. J. Anat. 91, 107-140.

PADYKULA, H. A., E. HERMAN (1955): Factors affecting the activity of adenosine triphosphatase, and other phosphatases as measured by histochemical technique. J. Histochem. Cytochem. 3, 161-169.

ROMEIS, B. (1968): Mikroskopische Technik. R. Ouldenbourg Verlag. München, Wien.

STAUN, H. (1972): The nutritional and genetic influence on number and size muscle fibers and their response to carcase quality in pigs. World review of Animal Production 8, 18.

Received: 8 June 1998
Accepted: 30 June 1998

JELIC, A., S. NEJEDLI, A. HRASTE, M. ZOBUNDZIJA, K. BOTKA PETRAK: Utjecaj dodatka enzima u hrani na histokemijske promjene u bijeloj i tamnoj piletini. Vet. arhiv 68, 143-148, 1998.

SAZETAK

U provedenim istrazivanjima koristeni su pilici linije "Jata". U pokusu je bilo 20 pilica podijeljenih u dvije skupine: kontrolnu i pokusnu s po 10 pilica u svakoj skupini. Tijekom cijelog perioda tova koji je trajao 42 dana pilici su bili hranjeni krmnom smjesom u kojoj je najveci udio cinio kukuruz. U pokusnoj skupini brojlera smo tijekom tova u hranu umijesali polienzimatski pripravak u kolicini od 0,5/1000 kg. Dodatak tog polienzimatskog preparata pilicima u hrani u ovoj kolicini doveo je do povecanja promjera vlakana u m. pectoralis superficialis i m. iliofibularis, te postotka bijelih brzokontrahirajucih glikolitickih vlakana u m. iliofibularis pilica pokusne skupine.

Kljucne rijeci: enzimi, pilici, histologija, m. iliofibularis, m. pectoralis superficialis, piletina