Probiotic Bacteria As Yoghurt Starter And Its Implication Effect To The Pathogenic And Non Pathogenic Bacteria In Mice Gastrointestinal.
PROBIOTIC BACTERIA AS YOGHURT STARTER AND
ITS IMPLICATION EFFECT TO THE PATHOGENIC AND NON
PATHOGENIC BACTERIA IN MICE GASTROINTESTINAL
Lovita Adriani, Hendronoto A.W. Lengkey
Faculty of Animal Husbandry, University of Padjadjaran, Bandung, INDONESIA
lovita_yoghurt@yahoo.co.id
The purpose of the research was to study the effect of bacteria consortium of Lactobacillus
bulgaricus, Streptococcus thermophillus, Lactobacillus acidophilus and Bifidobacteria on the
ecosystem of gastrointestinal in mice.The aim of this study was to explore the differences
between yoghurt content from different consortium, with 1,25% dosage; in mice during
three until five weeks. Also, the effect on the number of population of non pathogenic
bacteria (Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus and
Bifidobacteria ) and the total pathogenic bacteria in the segment of the mice gut (jejunum,
ilium and colon). Results indicated that the bacteria mixture have a good implementation in
microbial intestine of mice, which increased the population of non pathogenic and decreased
of pathogenic bacteria.
Keywords: total numbers of bacteria, pathogenic bacteria, and non‐pathogenic bacteria.
According to the previous studies, some genera of lactic acid bacteria and bifidobacteria
make an extremely important group of probiotic bacteria. Microflora of the gastrointestinal
tract of human or animal, they offer considerable potensial as probiotic because of their
history of save use and the general body of evidence that supports their positives role
(Björkstén, et al, 2001, Guarner and Malagelada, 2003a, Sears,2005 and Steinhoff , 2005).
Namely probiotics are microorganism which had been included in food without any adverse
effects and which were present in the gastrointestinal tract for health. At present, these
microorganism, called probiotics, have been selected from mostly lactic acid bacteria, e.g.
Lactobacillus acidophilus. Bifidobacteria is a part of the normal intestinal microflora of
human, since the microorganism are indigenous to the colon. The importance of an
indigenous microflora in the gastrointestinal tract as a natural resistance factor against
potential pathogenic microorganism was already recognised by Metchnicoff. Probiotic strain
can be used only , if the microorganism active in the body of the host if the fulfill a large
number of criteria. On the other side, lactic acid and acetic acid caused intestine acidity and
can prevented the growth of pathogenic bacteria. Those acids reduce absorbsion of
ammonia and amine since the large number of ammonia and amine can rise blood pressure,
cholesterol,and cancer because of nitrosamine.
Lactobacillus is a group of gram‐positive anaerobic colonic bacteria that produce lactic acid.
Supplementation with Lactobacillus and the resultant increased colonic levels of this
organism has maintained the health of subjects with several intestinal disorders including
diarrhea, ulcerative colitis (Bettelheim, et al, 1974 and Tap, et al, 2009). Research has shown
that Lactobacillus effectively competes with pathogenic bacteria in the colon for binding to
the epithelial cells that line the intestines (Schwiertz, 2003). It also inhibits pathogenic
bacteria by producing lactic acid and increasing epithelial mucous production.
262
Lucrări Științifice – vol 53 seria Medicină Veterinară
MATERIALS AND METHODS
Raw milk used for making yoghurt, from farm animal at the faculty. The bacteria are pure
cultivated Bifidobacterium, Lactobacillus acidophilus, Lactobacillus bulgaricus and
Streptococcus thermophilus. The identification of the cultures was based on the
characteristics of lactobacilli and streptococci as described in Bergey’s Manual of
Determinative Bacteriology (Holt et al, 1994).
120 mice, from a commercial hatchery.
The experimental design were Completely Randomized Design (CRD), with six treatment and
four replication. Every cages was filled with five mice.
RESULTS AND DISCUSSION
Total non pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal (cfu/ml).
In Table 1, presents the total non pathogenic bacteria in each segment of 6 weeks
mice gastro intestinal (cfu/ml).
In Table 1 , the total of non pathogenic bacteria in the colon are higher than the total
bacteria in the stomach, jejunum and ileum, especially with ration + Bifidobacteria starter
(R3, R4 and R5). Helpful bacteria prevent the growth of pathogenic species by competing for
nutrition and attachment sites to the epithelium of the colon. Symbiotic bacteria are more
at home in this ecological niche and are thus more successful in the competition. Indigenous
gut floras also produce bacteriocins which are proteinacious toxins that inhibit growth of
similar bacterial strains, substances which kill harmful microbes and the levels of which can
be regulated by enzymes produced by the host (Guarner and Malagelada, 2003a). The
resident gut microflora positively control the intestinal epithelial cell differentiation and
proliferation through the production of short‐chain fatty acids. They also mediate other
metabolic effects such as the syntheses of vitamins like biotin and folate as well as
absorption of ions including Magnesium, Calcium and Iron (O'Hara and Shanahan , 2006).
263
Lucrări Științifice – vol 53 seria Medicină Veterinară
264
Universitatea de Științe Agricole și Medicină Veterinară Iași
Notes :
Lb = Lactobacillus bulgaricus,
La = Lactobacillus acidophilus
R0 = Controle ration (without yoghurt)
R2 = Controle ration + Lb, St and La starter
R4 = Controle ration + Lb, St, La and B starter
St = Streptococcus thermophilus,
B = Bifidobacterium spp.
R1 = Controle ration + Lb and St starter
R3 = Controle ration + Lb, St and B starter
R5 = Controle ration + La and B starter
Total pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal (cfu/ml).
In Table 2, presents the total pathogenic bacteria in each segment of 6 weeks mice
gastro intestinal (cfu/ml).
Tabel 2. Total pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal
Notes :
B = Bacillus,
Mcc = Micrococci,
R0 = Controle ration (without yoghurt)
R2 = Controle ration + Lb, St and La starter
R4 = Controle ration + Lb, St, La and B starter
Stp = Staphylococcus aureus.
R1 = Controle ration + Lb and St starter
R3 = Controle ration + Lb, St and B starter
R5 = Controle ration + La and B starter
From the data from Table 2, the mice that have been feed with R5, has reducing the
pathogenic bacteria especially in the colon. This fact are agree with Beaugerie and Petit
(2004) that fermentation process, since it produces lactic acid and different fatty acids, also
serves to lower the pH in the colon, preventing the proliferation of harmful species of
bacteria and facilitating that of helpful species. The pH may also enhance the excretion of
carcinogens.
Metabolic function
The gut flora plays a major role in metabolizing dietary carcinogens (Junjie Qin; et al (2009);
the microcomponents and the macrocomponents. The microcomponents are genotoxic and
the major focus is on recent advances in heterocyclic amines (HCAs) which are produced by
cooking proteinaceous food such as meat and fish which can then induce tumors in organs
like breast, colon and prostate. HCAs are naturally occurring therefore the complete
avoidance ofthem is impractical which is why the metabolic function of gut flora of such
components is of
265
Lucrări Științifice – vol 53 seria Medicină Veterinară
great importance to our body as this would help in prevention of such tumors that are
difficult to avoid. The macrocomponents consists of the excessive intake of fat and sodium
chloride which can later promote tumors such as in breasts and colons from fat and gastric
carcinogenesis from sodium chloride(Guarner and Malagelada , 2003b)
CONCLUSION
The yoghurt with consortium starters with Lactobacillus acidophilus and Bifidobacterium
spp. with 1.25% dosage of mice body weight have been raised the non pathogenic
population and decreased the pathogenic bacteria in colon.
BIBLIOGRAPHY
Beaugerie L, Petit JC (2004). "Microbial‐gut interactions in health and disease. Antibiotic‐associated
diarrhoea". Best Pract Res Clin Gastroenterol 18 (2): 337–52. doi:10.1016/j.bpg.2003.10.002.
2. PMID 15123074. http://linkinghub.elsevier.com/retrieve/pii/S1521691803001276
3. Bettelheim KA, Breadon A, Faiers MC, O'Farrell SM, Shooter RA (1974). "The origin of O serotypes of
Escherichia coli in babies after normal delivery". J Hyg (Lond) 72 (1): 67–70. PMID 4593741.
4. Björkstén B, Sepp E, Julge K, Voor T, Mikelsaar M (2001). "Allergy development and the intestinal
microflora during the first year of life". J. Allergy Clin. Immunol. 108 (4): 516–
20.doi:10.1067/mai.2001.118130.
PMID
11590374.
http://linkinghub.elsevier.com/retrieve/pii/S0091‐ 6749(01)96140‐8
5. Guarner F, Malagelada JR (2003a). "Gut flora in health and disease". Lancet 361 (9356): 512–9.
doi:10.1016/S0140‐6736(03)12489‐0.
PMID
12583961.
http://linkinghub.elsevier.com/retrieve/pii/S0140‐6736(03)12489‐0
6. Guarner F, Malagelada JR (2003b). "Role of bacteria in experimental colitis". Best Pract Res Clin
Gastroenterol 17 (5): 793–804. doi:10.1016/S1521‐6918(03)00068‐4. PMID 14507589.
http://linkinghub.elsevier.com/retrieve/pii/S1521691803000684
7. Holt, J.G., N.R. Krieg, P.H.A. Sneath, J. T. Staley, and S. T. Williams, 1994. Bergey’s Manual of
determination Bacteriology, 9th ed. Williams and Williams. Baltimore. p. 566.
8. Junjie Qin; et al (2009), "A human gut microbial gene catalogue established by metagenomic
sequencing",
Nature
(464):
59–65,
doi:10.1038/nature08821,
http://www.nature.com/nature/journal/v464/n7285/full/nature08821.html, retrieved 2010‐03‐06
9. O'Hara AM, Shanahan F (2006). "The gut flora as a forgotten organ". EMBO Rep. 7 (7): 688–93.
doi:10.1038/sj.embor.7400731. PMID 16819463. Schwiertz A, Gruhl B, Löbnitz M, Michel P, Radke M,
Blaut M (2003). "Development of the intestinal bacterial composition in hospitalized preterm infants
in comparison with breast‐fed, full‐term infants". Pediatr. Res. 54 (3): 393–9.
doi:10.1203/01.PDR.0000078274.74607.7A.
PMID
12788986.
http://meta.wkhealth.com/pt/pt‐core/templatejournal/
lwwgateway/media/landingpage.htm?issn=0031‐3998&volume=54&issue=3&spage=393
10. Sears CL (2005). "A dynamic partnership: celebrating our gut flora". Anaerobe 11 (5): 247–51.
doi:10.1016/j.anaerobe.2005.05.001.
PMID
16701579.
http://linkinghub.elsevier.com/retrieve/pii/S1075‐9964(05)00068‐5
1. Steinhoff U (2005). "Who controls the crowd? New findings and old questions about the intestinal
microflora". Immunol. Lett. 99 (1): 12–6. doi:10.1016/j.imlet.2004.12.013. PMID 15894105.
http://linkinghub.elsevier.com/retrieve/pii/S0165‐2478(05)00005‐2
2. Tap J, Mondot S, Levenez F, Pelletier E, Caron C, Furet JP, Ugarte E, Muñoz‐Tamayo R, Paslier DL, Nalin
R, Dore J, Leclerc M (2009). "Towards the human intestinal microbiota phylogenetic core.". Env.
Microbiol. 11 (10): 2574–2584. doi:10.1111/j.1462‐2920.2009.01982.x. PMID 19601958.
http://www.ncbi.nlm.nih.gov/pubmed/19601958
1.
266
ITS IMPLICATION EFFECT TO THE PATHOGENIC AND NON
PATHOGENIC BACTERIA IN MICE GASTROINTESTINAL
Lovita Adriani, Hendronoto A.W. Lengkey
Faculty of Animal Husbandry, University of Padjadjaran, Bandung, INDONESIA
lovita_yoghurt@yahoo.co.id
The purpose of the research was to study the effect of bacteria consortium of Lactobacillus
bulgaricus, Streptococcus thermophillus, Lactobacillus acidophilus and Bifidobacteria on the
ecosystem of gastrointestinal in mice.The aim of this study was to explore the differences
between yoghurt content from different consortium, with 1,25% dosage; in mice during
three until five weeks. Also, the effect on the number of population of non pathogenic
bacteria (Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus and
Bifidobacteria ) and the total pathogenic bacteria in the segment of the mice gut (jejunum,
ilium and colon). Results indicated that the bacteria mixture have a good implementation in
microbial intestine of mice, which increased the population of non pathogenic and decreased
of pathogenic bacteria.
Keywords: total numbers of bacteria, pathogenic bacteria, and non‐pathogenic bacteria.
According to the previous studies, some genera of lactic acid bacteria and bifidobacteria
make an extremely important group of probiotic bacteria. Microflora of the gastrointestinal
tract of human or animal, they offer considerable potensial as probiotic because of their
history of save use and the general body of evidence that supports their positives role
(Björkstén, et al, 2001, Guarner and Malagelada, 2003a, Sears,2005 and Steinhoff , 2005).
Namely probiotics are microorganism which had been included in food without any adverse
effects and which were present in the gastrointestinal tract for health. At present, these
microorganism, called probiotics, have been selected from mostly lactic acid bacteria, e.g.
Lactobacillus acidophilus. Bifidobacteria is a part of the normal intestinal microflora of
human, since the microorganism are indigenous to the colon. The importance of an
indigenous microflora in the gastrointestinal tract as a natural resistance factor against
potential pathogenic microorganism was already recognised by Metchnicoff. Probiotic strain
can be used only , if the microorganism active in the body of the host if the fulfill a large
number of criteria. On the other side, lactic acid and acetic acid caused intestine acidity and
can prevented the growth of pathogenic bacteria. Those acids reduce absorbsion of
ammonia and amine since the large number of ammonia and amine can rise blood pressure,
cholesterol,and cancer because of nitrosamine.
Lactobacillus is a group of gram‐positive anaerobic colonic bacteria that produce lactic acid.
Supplementation with Lactobacillus and the resultant increased colonic levels of this
organism has maintained the health of subjects with several intestinal disorders including
diarrhea, ulcerative colitis (Bettelheim, et al, 1974 and Tap, et al, 2009). Research has shown
that Lactobacillus effectively competes with pathogenic bacteria in the colon for binding to
the epithelial cells that line the intestines (Schwiertz, 2003). It also inhibits pathogenic
bacteria by producing lactic acid and increasing epithelial mucous production.
262
Lucrări Științifice – vol 53 seria Medicină Veterinară
MATERIALS AND METHODS
Raw milk used for making yoghurt, from farm animal at the faculty. The bacteria are pure
cultivated Bifidobacterium, Lactobacillus acidophilus, Lactobacillus bulgaricus and
Streptococcus thermophilus. The identification of the cultures was based on the
characteristics of lactobacilli and streptococci as described in Bergey’s Manual of
Determinative Bacteriology (Holt et al, 1994).
120 mice, from a commercial hatchery.
The experimental design were Completely Randomized Design (CRD), with six treatment and
four replication. Every cages was filled with five mice.
RESULTS AND DISCUSSION
Total non pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal (cfu/ml).
In Table 1, presents the total non pathogenic bacteria in each segment of 6 weeks
mice gastro intestinal (cfu/ml).
In Table 1 , the total of non pathogenic bacteria in the colon are higher than the total
bacteria in the stomach, jejunum and ileum, especially with ration + Bifidobacteria starter
(R3, R4 and R5). Helpful bacteria prevent the growth of pathogenic species by competing for
nutrition and attachment sites to the epithelium of the colon. Symbiotic bacteria are more
at home in this ecological niche and are thus more successful in the competition. Indigenous
gut floras also produce bacteriocins which are proteinacious toxins that inhibit growth of
similar bacterial strains, substances which kill harmful microbes and the levels of which can
be regulated by enzymes produced by the host (Guarner and Malagelada, 2003a). The
resident gut microflora positively control the intestinal epithelial cell differentiation and
proliferation through the production of short‐chain fatty acids. They also mediate other
metabolic effects such as the syntheses of vitamins like biotin and folate as well as
absorption of ions including Magnesium, Calcium and Iron (O'Hara and Shanahan , 2006).
263
Lucrări Științifice – vol 53 seria Medicină Veterinară
264
Universitatea de Științe Agricole și Medicină Veterinară Iași
Notes :
Lb = Lactobacillus bulgaricus,
La = Lactobacillus acidophilus
R0 = Controle ration (without yoghurt)
R2 = Controle ration + Lb, St and La starter
R4 = Controle ration + Lb, St, La and B starter
St = Streptococcus thermophilus,
B = Bifidobacterium spp.
R1 = Controle ration + Lb and St starter
R3 = Controle ration + Lb, St and B starter
R5 = Controle ration + La and B starter
Total pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal (cfu/ml).
In Table 2, presents the total pathogenic bacteria in each segment of 6 weeks mice
gastro intestinal (cfu/ml).
Tabel 2. Total pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal
Notes :
B = Bacillus,
Mcc = Micrococci,
R0 = Controle ration (without yoghurt)
R2 = Controle ration + Lb, St and La starter
R4 = Controle ration + Lb, St, La and B starter
Stp = Staphylococcus aureus.
R1 = Controle ration + Lb and St starter
R3 = Controle ration + Lb, St and B starter
R5 = Controle ration + La and B starter
From the data from Table 2, the mice that have been feed with R5, has reducing the
pathogenic bacteria especially in the colon. This fact are agree with Beaugerie and Petit
(2004) that fermentation process, since it produces lactic acid and different fatty acids, also
serves to lower the pH in the colon, preventing the proliferation of harmful species of
bacteria and facilitating that of helpful species. The pH may also enhance the excretion of
carcinogens.
Metabolic function
The gut flora plays a major role in metabolizing dietary carcinogens (Junjie Qin; et al (2009);
the microcomponents and the macrocomponents. The microcomponents are genotoxic and
the major focus is on recent advances in heterocyclic amines (HCAs) which are produced by
cooking proteinaceous food such as meat and fish which can then induce tumors in organs
like breast, colon and prostate. HCAs are naturally occurring therefore the complete
avoidance ofthem is impractical which is why the metabolic function of gut flora of such
components is of
265
Lucrări Științifice – vol 53 seria Medicină Veterinară
great importance to our body as this would help in prevention of such tumors that are
difficult to avoid. The macrocomponents consists of the excessive intake of fat and sodium
chloride which can later promote tumors such as in breasts and colons from fat and gastric
carcinogenesis from sodium chloride(Guarner and Malagelada , 2003b)
CONCLUSION
The yoghurt with consortium starters with Lactobacillus acidophilus and Bifidobacterium
spp. with 1.25% dosage of mice body weight have been raised the non pathogenic
population and decreased the pathogenic bacteria in colon.
BIBLIOGRAPHY
Beaugerie L, Petit JC (2004). "Microbial‐gut interactions in health and disease. Antibiotic‐associated
diarrhoea". Best Pract Res Clin Gastroenterol 18 (2): 337–52. doi:10.1016/j.bpg.2003.10.002.
2. PMID 15123074. http://linkinghub.elsevier.com/retrieve/pii/S1521691803001276
3. Bettelheim KA, Breadon A, Faiers MC, O'Farrell SM, Shooter RA (1974). "The origin of O serotypes of
Escherichia coli in babies after normal delivery". J Hyg (Lond) 72 (1): 67–70. PMID 4593741.
4. Björkstén B, Sepp E, Julge K, Voor T, Mikelsaar M (2001). "Allergy development and the intestinal
microflora during the first year of life". J. Allergy Clin. Immunol. 108 (4): 516–
20.doi:10.1067/mai.2001.118130.
PMID
11590374.
http://linkinghub.elsevier.com/retrieve/pii/S0091‐ 6749(01)96140‐8
5. Guarner F, Malagelada JR (2003a). "Gut flora in health and disease". Lancet 361 (9356): 512–9.
doi:10.1016/S0140‐6736(03)12489‐0.
PMID
12583961.
http://linkinghub.elsevier.com/retrieve/pii/S0140‐6736(03)12489‐0
6. Guarner F, Malagelada JR (2003b). "Role of bacteria in experimental colitis". Best Pract Res Clin
Gastroenterol 17 (5): 793–804. doi:10.1016/S1521‐6918(03)00068‐4. PMID 14507589.
http://linkinghub.elsevier.com/retrieve/pii/S1521691803000684
7. Holt, J.G., N.R. Krieg, P.H.A. Sneath, J. T. Staley, and S. T. Williams, 1994. Bergey’s Manual of
determination Bacteriology, 9th ed. Williams and Williams. Baltimore. p. 566.
8. Junjie Qin; et al (2009), "A human gut microbial gene catalogue established by metagenomic
sequencing",
Nature
(464):
59–65,
doi:10.1038/nature08821,
http://www.nature.com/nature/journal/v464/n7285/full/nature08821.html, retrieved 2010‐03‐06
9. O'Hara AM, Shanahan F (2006). "The gut flora as a forgotten organ". EMBO Rep. 7 (7): 688–93.
doi:10.1038/sj.embor.7400731. PMID 16819463. Schwiertz A, Gruhl B, Löbnitz M, Michel P, Radke M,
Blaut M (2003). "Development of the intestinal bacterial composition in hospitalized preterm infants
in comparison with breast‐fed, full‐term infants". Pediatr. Res. 54 (3): 393–9.
doi:10.1203/01.PDR.0000078274.74607.7A.
PMID
12788986.
http://meta.wkhealth.com/pt/pt‐core/templatejournal/
lwwgateway/media/landingpage.htm?issn=0031‐3998&volume=54&issue=3&spage=393
10. Sears CL (2005). "A dynamic partnership: celebrating our gut flora". Anaerobe 11 (5): 247–51.
doi:10.1016/j.anaerobe.2005.05.001.
PMID
16701579.
http://linkinghub.elsevier.com/retrieve/pii/S1075‐9964(05)00068‐5
1. Steinhoff U (2005). "Who controls the crowd? New findings and old questions about the intestinal
microflora". Immunol. Lett. 99 (1): 12–6. doi:10.1016/j.imlet.2004.12.013. PMID 15894105.
http://linkinghub.elsevier.com/retrieve/pii/S0165‐2478(05)00005‐2
2. Tap J, Mondot S, Levenez F, Pelletier E, Caron C, Furet JP, Ugarte E, Muñoz‐Tamayo R, Paslier DL, Nalin
R, Dore J, Leclerc M (2009). "Towards the human intestinal microbiota phylogenetic core.". Env.
Microbiol. 11 (10): 2574–2584. doi:10.1111/j.1462‐2920.2009.01982.x. PMID 19601958.
http://www.ncbi.nlm.nih.gov/pubmed/19601958
1.
266