EFFECT OF ADDITION OF FEED ANTIBIOTIC OR PROBIOTIC ON PERFORMANCE AND COMPOSITION OF INTESTINAL MICROFLORA OF PIGS

Anna Rekiel¹, Julita Gajewska², Justyna Więcek¹, Anita Miszczyk^2
1 Department of Animal Breeding and Production, Warsaw Agricultural University, Poland
² Department of Soil Environment Sciences, Warsaw Agricultural University, Poland

ABSTRACT

In the individual two-stage system, being ended with the slaughter and slaughter evaluation of 32 crossbred pigs, the complete mixtures with the addition of antibiotic flavomycin or probiotic (strain of Pedicoccus acidilactici bacteria MA18/5M) (during the first fattening stage) were employed. Post-mortem quantitative and qualitative bacteriological tests and quantitative mycological tests on the determination of the contents of duodenum, jejunum, ileum and large intestine were carried out. The comparable results of fattening and slaughter evaluation of pigs in the groups were obtained. After administration of probiotic, the number of acidifying bacteria was increased and that of Enterobacteriaceae family decreased. The application of biostimulator in fattening occurred to be favourable from the production point of view and seems to be an suitable alternative to feed antibiotics.

Key words: growing-finishing pigs, feed additives, fattening, slaughter value, intestinal microflora.

INTRODUCTION

The quantity and proportions of micro-organisms living in the alimentary tract are relatively constant and typical of the particular periods of life of individual. They are s/ubject to changes, depending on the consumed feeds, inter alias, on feed additives, in the situation of health as well as during disease and stress [14].

The subject of the studies is to determine the suitability and effectiveness of additives, replacing antibiotic growth stimulators [AGS] [6, 11, 17, 18]. In the light of the so-far conducted experiments, the effectiveness of employing the probiotics instead of AGS is, however, not univocal [5, 7].

The aim of the studies was to determine the effect of the feed antibiotic additive or its replacer, probiotic, on production results and composition of intestinal microflora of fatteners.

MATERIALS AND METHODS

The observations covered 32 three-breed crossbred pigs (Polish Large White x Polish Landrace) x Duroc and (PLW x PL) x Belgian Landrace (1:1), gilts and barrows (1:1). The animals were allocated to control (C) and experimental (E) groups by the method of analogues. The participation of animals of two genotypes and two genders was equal to 1:1.

The fatteners were managed and fed individually. In two-stage fattening, from body weight of ca 21 kg up to 56 kg and from 56 kg up to 100 kg, the full-ration mixtures were administrated. The composition of feed included ground barley, wheat meal and extraction soy meal, meat-bone meal, premix and L-lysine and DL-methionine. During the 1st and the 2nd period of fattening, the mixtures contained 12.3 and 12.2 MJ EM and 165-168 and 143-149 g of crude protein in 1 kg of the mixture, respectively. The diets for fatteners of C group contained 5% addition of premix with antibiotic – flavomycin and the experimental fatteners received premix without antibiotic. Group E received 0.01% addition of probiotic (strain of Pedio-coccus acidilactici bacteria MA18/5M).

Chemical analysis of raw materials and mixtures was conducted by AOAC method [1] with the application of Tecator apparatus.

After completion of fattening and slaughter, the abbreviated dissection was carried out [12].

After slaughter, fragments of intestines were collected from 6 fatteners from each group (10 cm each). In quantitative and qualitative bacteriological analyses and in quantitative mycological tests, the contents of the following parts of small intestine were utilised: duodenum (a), jejunum (b), ileum (c) and of the large intestine (d). The following media were used in bacteriological analyses: nutritive agar, nutritive agar with the 10-% addition of defibrinated sheep blood prepared acc. to McConkey, acc. to Baird – Parker, acc. to King B, acc. to Mueller – Hinton; lactose with bromocresol purple acc. to Eijkman, Rogosa, Mitchell, Wisman with the addition of 5% tomato juice and acc. to Wilson – Blair. In mycological tests, media acc. to Martin et al. and acc. to Sabouraudn were employed. The quantitative tests were performed by the method of surface inoculation (the method of “touched” plates). To count the colonies-forming units (CFU), the counter of Stuart Scientific company was used. In qualitative analyses, the reducing surface inoculation (the scratch method) was applied.The quality evaluation was conducted on the ground of capability to grow, observation of macroscopic colonies and microscopic cells (preparations stained by the method acc. to Gram as well as and life performance preparations). The diagnostics of bacteria was carried out, based on the taxonomy of organisms acc. to Bergey’s Manual of Determinative Bacteriology [2].

The results were statistically analysed by single-factor analysis of variance, employing the least square method [16].

RESULTS AND DISCUSSION

In the studies on intake and conversion of feed, growth rate and the chosen slaughter traits, any statistically significant differences were not found between the groups of fatteners, receiving the different feed additives (Table 1). The total feed intake in fattening period was higher in group E as compared to group C and feed conversion was worse by 1.03% respectively. The experimental fatteners as compared to the control ones grew slower by 1.91% (E). In case of the application of probiotics Jasek et al [5], Kumprecht and Zobač [9], Sokół et al [15] found the improvement of the growth rate and feed conversion by the growing pigs. A small improvement of slaughter traits of the pigs, receiving the addition of biostimulator in their diet (group E) as compared to group C has been confirmed by studies of Doyle [3], Grela et al [4], Jasek et al [5].

Table 1. Production results of pigs used in the experiment

Specification	Control	Experimental	Se
Total feed intake: fattening period I, kg	81.80	82.80	3.03
fattening period II, kg	133.90	139.50	3.98
during fattening from 21 to 100 kg, kg	215.70	222.40	5.25
Feed conversion per 1 kg of BW gain: during fattening period I, kg	2.35	2.34	0.069
during fattening period II, kg	3.04	3.22	0.097
mean value during fattening period from 21 to 100 kg, kg	2.73	2.82	0.067
Daily gains: fattening period I, g	723	731	17.40
fattening period II, g	853	808	23.60
mean value during fattening period from 21 to 100 kg, g	786	771	17.98
Dressing percentage, %	75.90	76.30	0.41
Mean thickness of backfat from 5 measurements, cm	2.26	2.27	0.09
Loin “eye” area, cm2	49.20	50.30	1.54
Meatiness of basic cuts, %	58.90	60.40	0.75

The number of bacteria from Enterobacteriaceae family and lactic acid bacteria (LAB) in 1 g of dry matter of the intestinal content of the fatteners in group C was equal to 10⁷-10⁸ (Table 2). In the contents of large intestine, the ratio of micro-organisms from the two mentioned above groups was also similar. In the contents of the intestines of fatteners from group E, the number of bacteria from Enterobacteriaceae family amounted to ca 10⁷ whereas the number of acidifying bacteria was higher than in group C (10⁹-10¹⁰ in 1 g of dry matter). The decrease of the number of bacteria from Enterobacteriaceae family in group E in relation to group C may be considered as phenomenon exclusively favourable. From among the types of Escherichia, the species Escherichia coli belongs to the mentioned above family. The most of its representatives reveals the opportunistic pathogenicity. They may cause infections of alimentary tract; the presence of genes, located in plasmid and determining generation of haemolysins in haemolytic E. coli strains affects a quick spreading out of infection [2].

Table 2. Number of the chosen groups of micro-organisms in the intestinal contents

Group	Intestine fragment	Number of the chosen groups of micro-organisms on different media, g-1, DM-1
		1	2	3	4	5	6	7
Control	a	1010	1.15 x 108	2.72 x 107	3.11 x 107	3.40 x 107	1.46 x 108	+
	b	109	1.14 x 108	4.60 x 107	1.46 x 108	2.30 x 106	1.46 x 108	+
	c	109	8.70 x 107	2.57 x 107	4.95 x 107	4.33 x 107	7.10 x 107	+
	d	108	1.63 x 108	2.43 x 108	3.17 x 107	1.15 x 107	1.19 x 107	+
Experimental	a	1010	2.63 x 107	1.18 x 109	2.70 x 108	2.40 x 106	2.70 x 108	+
	b	1010	2.17 x 107	3.48 x 109	1.29 x 108	5.82 x 107	1.29 x 108	+
	c	1010	1.05 x 107	4.21 x 1010	2.96 x 108	1.60 x 106	2.95 x 108	+
	d	1010	1.55 x 107	5.60 x 1010	2.69 x 109	2.27 x 106	2.69 x 109	+

Fragment of intestines: a – duodenum, b – jejunum, c – ileum, d – large intestine. The number of the chosen groups of micro-organisms grown on medium: 1. total acc. to Mueller-Hinton, 2. total from Enterobacteriaceae family acc. to McConkey, 3. total acidifying bacteria number – lactose with bromocresol purple acc. to Eijkman, 4. total of acidifying bacteria acc. to Rogosa, Mitchell, Wiesman with the addition of 5% tomato juice, 5. total number of yeasts and microscopic hypha fungi acc. to Martin, 6. number of Proteus vulgaris bacteria on nutritive agar, 7. presence of Clostridium perfringens bacteria on medium acc. to Wilson-Blair. Determination of bacterial growth: + growth, – lack of growth.

By “occupation of the place”, the probiotics exclude or reduce adherence of pathogens, Escherichia coli, Salmonella sp. They produce bacteriocins, being antagonistic for their growth, lower acidity and increase the effect of digestive enzymes. The probiotic Pediococcus bacteria from Bactocell preparation are also characterized by capability of co-aggregating. Probiotics decide on the composition of the intestinal microflora of pigs and affect favourably the host’s organism [13, 18].

In group E as compared to group C, bacteria from LAB group were 1000-times more numerous. They had probably an influence on the 10-100 th fold decrease of the number of bacteria from Enterobacteriaceae family in relation to group C. The antagonistic metabolites, produced by autochthonous lactic acid bacteria include, among others, bacteriocins. They are active in relation to Gram-negative bacteria, e.g. Clostridium. By generation of pores in cytoplasmic membrane of sensitive bacteria and outflow of electrolytes and low-molecular metabolites, they cause the death of a cell [8]. Clostridium perfringens produces toxins which affect the secretion activity of intestine, increase a difference between potentials in ions’ transport, damage the cells or tissues, change the activity of unstriated muscles in the intestines [10]. The presence of Clostridium perfringens in the contents of fatteners’ intestines was found more frequently in group C than in group E. The limitation of their number decre-ased their participation in competition for nutrients and was favourable for better productivity of the experimental animals.

A high similarity of the composition of intestinal microflora in fatteners from the two examined groups was observed (Table 3). Only in group E, bacteria from Bacillus and Pediococcus type were found. The presence of bacteria from Pediococcus type resulted probably from the addition of probiotic preparation, containing P. acidilactici strain MA 18/5M, to the diet. Aerobic cocci, Gram-positive Pediococcus present in the mentioned preparation, gain energy in reactions of glucose and lactose fermentation. They are micro-aero-tolerant. Stability of resident microflora may be increased by administration of excluding – competitive cultures or feeding with probiotic compounds.

Table 3. Micro-organisms isolated from the examined material

Group	Isolated micro-organisms – type
Control	Escherichia, Pseudomonas, Streptococcus, Enterococcus, Lactobacillus, Lactococcus, Staphylococcus, Proteus, Clostridium, yeasts, microscopic hypha fungi
Experimental	Escherichia, Pseudomonas, Streptococcus, Enterococcus, Pediococcus, Lactobacillus, Lactococcus, Staphylococcus, Proteus, Clostridium, Bacillus, yeasts, microscopic hypha fungi

The absence of pathogenic ß-haemolytic strains: Escherichia coli and Salmonella sp. in ali-mentary tract of the examined animals of the two feeding groups was stated; it was an evidence of their good health state.

CONCLUSIONS

1. When applying the addition of feed antibiotic – flavomycin or, alternatively, of probiotic (strain of Pedicoccus acidilactici bacteria MA18/5M) in the diets for growing pigs, the comparable results of fattening and slaughter evaluation were obtained.

2. The application of probiotic (group E) caused the increase of acidifying bacteria number and decreased the number of bacteria from Enterobacteriaceae family.

3. The alternative use of biostimulator instead of feed antibiotic occurred to be favourable from the production viewpoint and seems to be the suitable alternative for antibiotics.

REFERENCES

1. AOAC, 1990. Association of Official Analitical Chemists. Official Methods of Analysis. 15th Edition. Arlington, VA.

2. Buchanan R.E., Gibbons N.E. (Ed.), 1974. Bergey’s Manual of Determinative Bacteriology. 8th ed., Williams and Wilkins Co., Baltimore

3. Doyle M.E., 2001. Alternatives to antibiotic use for growth promotion in animal husbandry. Food Research Institute, 1-17.

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6. Kirchgatterer A., Knoflach P., 2004. Natural therapy instead of chemistry? Probiotics in gastroenterology. Acta Med. Austriaca 31(1), 13-17.

7. Kornegay E.T., Risley C.R., 1996. Nutrient digestibilities of a cornsoybean meal diet as influenced by Bacillus products fed to finishing swine. J. Anim. Sci. 74, 779-805.

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9. Kumprecht I., Zobač P., 1998. Study of the effect of a combined preparation containing Enterococcus faecium M-74 and mannan oligosaccharides in diets for weanling piglets. Czech J. Anim. Sci. 43, 477-481.

10. Niemiałtowski M., Toka F.N., 1996. Bakterie jelitowe i ich toksyny – od mikroskopu do biologii molekularnej [w: Choroby zakazne i inwazyjne przewodu pokarmowego ludzi i zwierzšt] [The bacteria and their toxins – from microscope to molecular biology. Infectious and invasive diseases of intestinal tract humen and animals]. Mateiały Konferencyjne. Kazimierz Dolny 24-26 paŸdziernika1996 [in Polish].

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12. Różycki M., 1996. Zasady postępowania przy ocenie swiń w Stacjach Kontroli Użytkowosci Rzeznej Trzody Chlewnej. Stan hodowli i wyniki oceny swiń [Procedures of pig evaluation at pig performance testing stations. State of breeding and evaluation results of swine]. IZ XIV, 69-82 [in Polish].

13. Schrezenmeir J., de Vrese M., 2001. Probiotics, prebiotics, and synbiotics – approaching a definition. Am. Society Clin. Nutr. 73, 361-364.

14. Stavric S., Kornegay E.T., 1995. Microbial probiotics for pigs and poultry. [in: Biotech-nology in Animal Feeds and Animal Feeding]. Ed. R.J. Wallace, A. Chesson: VCH Verlagsgesellschaft mbH, Wienheim, 206-231.

15. Sokół J., Karas J., Stecka K., Bobel B.K., Grzybowski R., 2001. Użytecznosc probiotyków wieloskładnikowych w żywieniu tuczników [Usability of multicomponen-tial probiotics in feeding of fatteners]. Anim. Sci. Special. number, 224-229 [in Polish].

16. SPSS 10.0 for Windows user’s guide, 2000 by SPSS Ins.

17. Verstegen M.W., Williams B.A., 2002. Alternatives to the use of antibiotics as growth promoters for monogastric animals. Anim. Biotechnol. 13(1), 113-127.

18. Zimmermann B., Bauer E., Mosenthin R., 2001. Pro- and prebiotics in pig nutrition – potential modulators of gut healthy? J. Anim. Feed Sci. 10, 47-56.

Supported by the State Committee for Scientific Research in Poland (Project no 3 PO6Z 026 25) and DS.

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Anna Rekiel
Department of Animal Breeding and Production,
Warsaw Agricultural University, Poland
Ciszewskiego 8, 02-786 Warszawa, Poland
Phone/fax: (48 22) 5936561/ 8530914
email: rekiel@alpha.sggw.waw.pl

Julita Gajewska
Department of Soil Environment Sciences,
Warsaw Agricultural University, Poland
Nowoursynowska 159, 02-776 Warszawa, Poland

Justyna Więcek
Department of Animal Breeding and Production,
Warsaw Agricultural University, Poland
Ciszewskiego 8, 02-786 Warszawa, Poland
Phone/fax: (48 22) 5936561/ 8530914

Anita Miszczyk
Department of Soil Environment Sciences,
Warsaw Agricultural University, Poland
Nowoursynowska 159, 02-776 Warszawa, Poland

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