Control of Staphylococcus aureus in Meat System by in situ and ex situ Bacteriocins from Lactobacillus sakei and Pediococcus spp.
The present study consisted of an applied test in meat
system to assess the effectiveness of three bio agents bacteriocinproducing
strains: Lm24: Lactobacillus sakei, Lm14and Lm25:
Pediococcus spp. Two tests were carried out: The ex situ test was
intended for three batches added with crude bacteriocin solutions at
12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the
in situ one consisted of four batches; three of them inoculated with
one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one
was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The
two used tests were done in the presence of the pathogen
St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch
served as a positive or a negative control was used too. The
incubation was performed at 7°C. Total viable counts, staphylococci
and lactic acid bacteria, at the beginning and at selected times with
interval of three days were enumerated. Physico-chemical
determinations (except for in situ test): pH, dry mater, sugars, fat and
total protein, at the beginning and at end of the experiment, were
done, according to the international norms. Our results confirmed the
ex situ effectiveness. Furthermore, the batches affected negatively the
total microbial load over the incubation days, and showed a
significant regression in staphylococcal load at day seven, for Lm14,
Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed
that, at the last day of culture, staphylococcal load was nil for the
three batches. In the in situ test, the cultures displayed less inhibitory
attitude and recorded a decrease in staphylococcal load, for Lm14,
Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore,
physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an
increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry
mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result
reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%,
0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%,
5.37%, 5.5%. This study suggests that the use of selected strains as
Lm25 could lead to the best results and would help in preserving and
extending the shelf life of lamb meat.
[1] U. Schillinger, and F.K. Lücke, "Identification of lactobacilli from meat
and meat products”, Food Microbiology.4, 1987, 199-208.
[2] Y. Morishita, and K. Shiromizu, "Characterization of lactobacilli
isolated from meats and meat products”, International Journal of Food
Microbiology, 3, 1986, 19-29.
[3] J. Samelis, F. Maurogenakis, and J. Metaxopoulos, "Characterisation of
lactic acid bacteria isolated from naturally fermented Greek dry salami”,
International Journal of Food Microbiology, 23, 1994, 179-196.
[4] A. Najjari, H. Ouzari, A. Boudabous, and M. Zagorec, "Method for
reliable isolation of Lactobacillus sakei strains originati from Tunisian
seafood and meat products”, International Journal of Food
Microbiology, 121, 2008, 342–351.
[5] R. Bromberg, I. Moreno, C.L. Zaganini, R.R. Delboni, and J. De
Oliveira, "Isolation of bacteriocin-producing lactic acid bacteria from
meat and meat products and its spectrum of inhibitory activity”,
Brazilian Journal of Microbiology, Vol.35, 2004.
[6] R. J. Jones, H. M. Hussein, M. Zagorec, G. Brightwell, and J. R Tagg,
"Isolation of lactic acid bacteria with inhibitory activity against
pathogens and spoilage organisms associated with fresh meat”, Food
Microbiology, Vol. 25, 2008.
[7] E. C. P. De-Martinis, and F .Z. Freitas, "Screening of lactic acid bacteria
from Brazilian meats for bacteriocin formation”, Food Control, Vol. 14,
2003.
[8] M. A. H. Al-Allaf, A. M. M. Al-Rawi, and A. T. Al-Mola,
"Antimicrobial activity of lactic acid bacteria isolated from minced beef
meat against some pathogenic bacteria”, Iraqi Journal of Veterinary
Sciences. Vol 23 Supplement I (2009) 115-117.
[9] J. Chaiyana, S. Boonrang, and S. Sinsuwongwat, "Isolation and
screening of bactériocin producing bacteria from fermented meat
products”, Biotechnology for gross national hapipiness, 2007, 245- 258.
[10] P. H. Castellano, W. H. Holzapfel,andG. M. Vignolo, "The control of
Listeria innocua and Lactobacillus sakei in broth and meat slurry with
the bacteriocinogenic strain Lactobacillus casei CRL705”, Food
Microbiology, 21, 2004, 291-298.
[11] C. Charlier, M. Cretenet, S. Even, and Y. Le Loir, "Interactions between
Staphylococcus aureus and lactic acid bacteria: An old story with new
perspectives”, International Journal of Food Microbiology. 13, 2009
30–39.
[12] A. Gálvez, H. Abriouel, R. L. López, and N. Ben Omar, "Bacteriocinbased
strategies for food biopreservation”, International Journal of Food
Microbiology. 120, 2007, 51-70.
[13] T. R. Klaenhammer, T. R. Barrangou, R. Buck, B. L. Azcarate-Peril, and
M. A. Altermann, "Genomic features of lactic acid bacteria effecting
bioprocessing and health”, FEMS Microbiol Rev, Vol. 29, 2005.
[14] P. Castellano, C. Belfiore, S. Fadda, and G. Vignolo, "A review of
bacteriocinogenic lactic acid bacteria used as bioprotective cultures in
fresh meat produced in Argentina”, Meat Science, Vol. 79, 2008.
[15] C. Dortu, and P. Thonart, "Les bactériocines des bactéries lactiques:
caractéristiques et intérêts pour la bioconservation des produits
alimentaires”, Biotechnol. Agron. Soc. Environ. 13, 2009.
[16] F. K Lucke, "Utilization of microbes to process and preserve meat”,
Meat Science, Vol.56, 2000, 105–115.
[17] W. H. Holzapfel, R. Geisen, and U. Schillinger, "Biological preservation
of foods with reference to protective cultures, bacteriocins and foodgrade
enzymes”, International Journal of Food Microbiology. 24, 1995,
343–36.
[18] T.R. Klaenhammer, "Bacteriocins of lactic acid bacteria”, Biochimie,
Vol. 70, 1988.
[19] T. Abee, L. Krockel, and C. Hill, "Bacteriocins : modes of action and
potentials in food preservation and control of food poisoning”,
International Journal of Food Microbiology, Vol. 28, 1995.
[20] A. S Naidu, R. Unal, and J. Tulpinski, "Bacteriocins: Antimicrobial
Activity and Applications”, In: K. Shetty, G. Paliyath, Anthony,
Pometto, and R. E. Levin, food biotechnology, 2006.
[21] T.R Klaenhammer, "Genetics of bacteriocins produced by lactic acid
bacteria”, Federation of European Microbiological Societies,
Microbiology Reviews, Vol. 12 1993.
[22] L. H. Deegana, P. D. Cottera, and C. Hilla, "Bacteriocine biological
tools for biopreservation”, International Dairy Journal, Vol.16, 2006,
1058–1071.
[23] M. Hugas, "Bacteriocinogenic Lactic Acid Bacteria for the
Biopreservation of Meat and Meat Products”, Meat Science. Vol 49,
1998, 139-150
[24] J. Guiraud, and P. Galzy, "L’analyse Microbiologique dans les industries
alimentaires”, L’usine, 1980, P235.
[25] M. E. Stiles, "Potential for biological control of agents of foodborne
disease”, Food Research International, Vol. 27, 1994.
[26] B. Ray, and A. bhunia, "Fundamental Food Microbiology”, 4 th edition,
CRC Press, 2008, 492 pp.
[27] I. Kalalou, M. Faid, and A. T. Ahami, "Extending shelf life of fresh
minced camel meat at ambient temperature by Lactobacillus dlbrueckii
subsp. Delbrueckii”, Electronic Journal of Biotechnology, Vol.7, 2004.
[28] S. Ananou, M. Maqueda, M. Martínez-Bueno, A. Gálvez, and
E. Valdivia, "Control of Staphylococcus aureus in sausages by enterocin
AS-48”, Meat Science. 71, 2005, 549–556.
[29] H. Khan, S. Flint, and P. L. Yu, "Enterocins in food preservation,
International Journal of Food Microbiology”, International Journal of
Food Microbiology, 2010.
[30] S. Ananou, M. Garriga, A. Jofré, T. Aymerich, A. Gálvez, M. Maqueda,
M. Martínez-Bueno, and E. Valdivia, "Combined effect of enterocin AS-
48 and high hydrostatic pressure to control food-borne pathogens
inoculated in low acid fermented sausages”, Meat Science, 2009.
[31] S. Ananou, M. Garriga, M. Hugas, M. Maqueda, M. Martínez-Bueno,
A. Gálvez, and E. Valdivia, "Control of Listeria monocytogenes in
model sausages by enterocin AS-48”, International Journal of Food
Microbiology, Vol.103, 2005, 179-190.
[32] L. V. Thomas, M. R. Clarkson, and J. Delves-Broughton, "Nisin”, in:
A. S. Naidu, Natural food antimicrobial systems, Boca Raton: CRC
Pres, 2000, 463– 524.
[33] G.F. De-Valdez, "Maintenance of Lactic Acid Bacteria”,in:
J. F. T. Spencer, and A. L. R. Spencer, Food Microbiology protocols,
Humana Press, 2001.
[34] A. Badis, N. Laouabdia-Sellami, D. Guetarni, M. Kihal, and R. Ouzrout,
"caracterisationphenotypique des bacteries lactiques isolees à partir de
lait cru de chèvre de deux populations caprines locales "arabia et
kabyle"”, Sciences et Technologie, Vol. 23, 2005.
[35] L. J. Harris, M. A. Daeschel, M. E. Stiles, and T. R. Klaenhammer,
"Antimicrobial activity of lactic acid bacteria against listeria
monocytogenes”, Journal of Food Protection, Vol. 52, 1989.
[36] U. Schillinger, and F.K. Lücke, "Antibacterial Activity of Lactobacillus
sake Isolated from Meat”, applied and environmental Microbiology, Vol.
55, 1989.
[37] S. A. Cuozzo, J. M. Sesma-Fernando, A. A. Holgado, R. Pesce, and
R. R. Raya, "Methods for the Detection and Concentration of
Bacteriocins Produced by Lactic Acid Bacteria”,in: F. T. S. John, and
L .R. S. Alicia, "Food Microbiology protocols”, Humana Press, 2001.
[38] A. Delgado, D. Brito, P. Fevereiro, R. Tenreiro, and C. Peres,
"Bioactivity quantification of crude bacteriocin solutions”, Journal of
Microbiological Methods, Vol. 62, 2005.
[39] G. Greer, and D. Dilts-Bryan, "Lactic acid inhibition of the growth of
spoilage bacteria and cold tolerant pathogens on pork”, International
Journal of Food Microbiology. 25, 1995, 141-151.
[40] J. Cleveland, T. J. Montville, I. F. Nes, and M. L. Chikindas,
"Bacteriocins: safe, natural antimicrobials for food preservation”,
International Journal of Food Microbiology. 71, 2001, 1–20.
[41] A. L. Vigil, E. Palou, M. E. Parish, and D. P. Michael, "Methods for
Activity Assay and Evaluation of Results”,in: D. P. Michael, J. N. Sofos,
and A. L. Branen, Antimicrobials in food , 2005, Third Edition, CRC
Press : 659- 680.
[42] Y. Babji, and T. R. K. Murthy, "Effect of inoculation of mesophilic
lactic acid bacteria on microbial and sensory changes of minced goat
meat during storage under vacuum and subsequent aerobic storage”,
Meat Science, 54, 2000, 197- 202.
[1] U. Schillinger, and F.K. Lücke, "Identification of lactobacilli from meat
and meat products”, Food Microbiology.4, 1987, 199-208.
[2] Y. Morishita, and K. Shiromizu, "Characterization of lactobacilli
isolated from meats and meat products”, International Journal of Food
Microbiology, 3, 1986, 19-29.
[3] J. Samelis, F. Maurogenakis, and J. Metaxopoulos, "Characterisation of
lactic acid bacteria isolated from naturally fermented Greek dry salami”,
International Journal of Food Microbiology, 23, 1994, 179-196.
[4] A. Najjari, H. Ouzari, A. Boudabous, and M. Zagorec, "Method for
reliable isolation of Lactobacillus sakei strains originati from Tunisian
seafood and meat products”, International Journal of Food
Microbiology, 121, 2008, 342–351.
[5] R. Bromberg, I. Moreno, C.L. Zaganini, R.R. Delboni, and J. De
Oliveira, "Isolation of bacteriocin-producing lactic acid bacteria from
meat and meat products and its spectrum of inhibitory activity”,
Brazilian Journal of Microbiology, Vol.35, 2004.
[6] R. J. Jones, H. M. Hussein, M. Zagorec, G. Brightwell, and J. R Tagg,
"Isolation of lactic acid bacteria with inhibitory activity against
pathogens and spoilage organisms associated with fresh meat”, Food
Microbiology, Vol. 25, 2008.
[7] E. C. P. De-Martinis, and F .Z. Freitas, "Screening of lactic acid bacteria
from Brazilian meats for bacteriocin formation”, Food Control, Vol. 14,
2003.
[8] M. A. H. Al-Allaf, A. M. M. Al-Rawi, and A. T. Al-Mola,
"Antimicrobial activity of lactic acid bacteria isolated from minced beef
meat against some pathogenic bacteria”, Iraqi Journal of Veterinary
Sciences. Vol 23 Supplement I (2009) 115-117.
[9] J. Chaiyana, S. Boonrang, and S. Sinsuwongwat, "Isolation and
screening of bactériocin producing bacteria from fermented meat
products”, Biotechnology for gross national hapipiness, 2007, 245- 258.
[10] P. H. Castellano, W. H. Holzapfel,andG. M. Vignolo, "The control of
Listeria innocua and Lactobacillus sakei in broth and meat slurry with
the bacteriocinogenic strain Lactobacillus casei CRL705”, Food
Microbiology, 21, 2004, 291-298.
[11] C. Charlier, M. Cretenet, S. Even, and Y. Le Loir, "Interactions between
Staphylococcus aureus and lactic acid bacteria: An old story with new
perspectives”, International Journal of Food Microbiology. 13, 2009
30–39.
[12] A. Gálvez, H. Abriouel, R. L. López, and N. Ben Omar, "Bacteriocinbased
strategies for food biopreservation”, International Journal of Food
Microbiology. 120, 2007, 51-70.
[13] T. R. Klaenhammer, T. R. Barrangou, R. Buck, B. L. Azcarate-Peril, and
M. A. Altermann, "Genomic features of lactic acid bacteria effecting
bioprocessing and health”, FEMS Microbiol Rev, Vol. 29, 2005.
[14] P. Castellano, C. Belfiore, S. Fadda, and G. Vignolo, "A review of
bacteriocinogenic lactic acid bacteria used as bioprotective cultures in
fresh meat produced in Argentina”, Meat Science, Vol. 79, 2008.
[15] C. Dortu, and P. Thonart, "Les bactériocines des bactéries lactiques:
caractéristiques et intérêts pour la bioconservation des produits
alimentaires”, Biotechnol. Agron. Soc. Environ. 13, 2009.
[16] F. K Lucke, "Utilization of microbes to process and preserve meat”,
Meat Science, Vol.56, 2000, 105–115.
[17] W. H. Holzapfel, R. Geisen, and U. Schillinger, "Biological preservation
of foods with reference to protective cultures, bacteriocins and foodgrade
enzymes”, International Journal of Food Microbiology. 24, 1995,
343–36.
[18] T.R. Klaenhammer, "Bacteriocins of lactic acid bacteria”, Biochimie,
Vol. 70, 1988.
[19] T. Abee, L. Krockel, and C. Hill, "Bacteriocins : modes of action and
potentials in food preservation and control of food poisoning”,
International Journal of Food Microbiology, Vol. 28, 1995.
[20] A. S Naidu, R. Unal, and J. Tulpinski, "Bacteriocins: Antimicrobial
Activity and Applications”, In: K. Shetty, G. Paliyath, Anthony,
Pometto, and R. E. Levin, food biotechnology, 2006.
[21] T.R Klaenhammer, "Genetics of bacteriocins produced by lactic acid
bacteria”, Federation of European Microbiological Societies,
Microbiology Reviews, Vol. 12 1993.
[22] L. H. Deegana, P. D. Cottera, and C. Hilla, "Bacteriocine biological
tools for biopreservation”, International Dairy Journal, Vol.16, 2006,
1058–1071.
[23] M. Hugas, "Bacteriocinogenic Lactic Acid Bacteria for the
Biopreservation of Meat and Meat Products”, Meat Science. Vol 49,
1998, 139-150
[24] J. Guiraud, and P. Galzy, "L’analyse Microbiologique dans les industries
alimentaires”, L’usine, 1980, P235.
[25] M. E. Stiles, "Potential for biological control of agents of foodborne
disease”, Food Research International, Vol. 27, 1994.
[26] B. Ray, and A. bhunia, "Fundamental Food Microbiology”, 4 th edition,
CRC Press, 2008, 492 pp.
[27] I. Kalalou, M. Faid, and A. T. Ahami, "Extending shelf life of fresh
minced camel meat at ambient temperature by Lactobacillus dlbrueckii
subsp. Delbrueckii”, Electronic Journal of Biotechnology, Vol.7, 2004.
[28] S. Ananou, M. Maqueda, M. Martínez-Bueno, A. Gálvez, and
E. Valdivia, "Control of Staphylococcus aureus in sausages by enterocin
AS-48”, Meat Science. 71, 2005, 549–556.
[29] H. Khan, S. Flint, and P. L. Yu, "Enterocins in food preservation,
International Journal of Food Microbiology”, International Journal of
Food Microbiology, 2010.
[30] S. Ananou, M. Garriga, A. Jofré, T. Aymerich, A. Gálvez, M. Maqueda,
M. Martínez-Bueno, and E. Valdivia, "Combined effect of enterocin AS-
48 and high hydrostatic pressure to control food-borne pathogens
inoculated in low acid fermented sausages”, Meat Science, 2009.
[31] S. Ananou, M. Garriga, M. Hugas, M. Maqueda, M. Martínez-Bueno,
A. Gálvez, and E. Valdivia, "Control of Listeria monocytogenes in
model sausages by enterocin AS-48”, International Journal of Food
Microbiology, Vol.103, 2005, 179-190.
[32] L. V. Thomas, M. R. Clarkson, and J. Delves-Broughton, "Nisin”, in:
A. S. Naidu, Natural food antimicrobial systems, Boca Raton: CRC
Pres, 2000, 463– 524.
[33] G.F. De-Valdez, "Maintenance of Lactic Acid Bacteria”,in:
J. F. T. Spencer, and A. L. R. Spencer, Food Microbiology protocols,
Humana Press, 2001.
[34] A. Badis, N. Laouabdia-Sellami, D. Guetarni, M. Kihal, and R. Ouzrout,
"caracterisationphenotypique des bacteries lactiques isolees à partir de
lait cru de chèvre de deux populations caprines locales "arabia et
kabyle"”, Sciences et Technologie, Vol. 23, 2005.
[35] L. J. Harris, M. A. Daeschel, M. E. Stiles, and T. R. Klaenhammer,
"Antimicrobial activity of lactic acid bacteria against listeria
monocytogenes”, Journal of Food Protection, Vol. 52, 1989.
[36] U. Schillinger, and F.K. Lücke, "Antibacterial Activity of Lactobacillus
sake Isolated from Meat”, applied and environmental Microbiology, Vol.
55, 1989.
[37] S. A. Cuozzo, J. M. Sesma-Fernando, A. A. Holgado, R. Pesce, and
R. R. Raya, "Methods for the Detection and Concentration of
Bacteriocins Produced by Lactic Acid Bacteria”,in: F. T. S. John, and
L .R. S. Alicia, "Food Microbiology protocols”, Humana Press, 2001.
[38] A. Delgado, D. Brito, P. Fevereiro, R. Tenreiro, and C. Peres,
"Bioactivity quantification of crude bacteriocin solutions”, Journal of
Microbiological Methods, Vol. 62, 2005.
[39] G. Greer, and D. Dilts-Bryan, "Lactic acid inhibition of the growth of
spoilage bacteria and cold tolerant pathogens on pork”, International
Journal of Food Microbiology. 25, 1995, 141-151.
[40] J. Cleveland, T. J. Montville, I. F. Nes, and M. L. Chikindas,
"Bacteriocins: safe, natural antimicrobials for food preservation”,
International Journal of Food Microbiology. 71, 2001, 1–20.
[41] A. L. Vigil, E. Palou, M. E. Parish, and D. P. Michael, "Methods for
Activity Assay and Evaluation of Results”,in: D. P. Michael, J. N. Sofos,
and A. L. Branen, Antimicrobials in food , 2005, Third Edition, CRC
Press : 659- 680.
[42] Y. Babji, and T. R. K. Murthy, "Effect of inoculation of mesophilic
lactic acid bacteria on microbial and sensory changes of minced goat
meat during storage under vacuum and subsequent aerobic storage”,
Meat Science, 54, 2000, 197- 202.
@article{"International Journal of Biological, Life and Agricultural Sciences:67417", author = "M. Naimi and M. B. Khaled", title = "Control of Staphylococcus aureus in Meat System by in situ and ex situ Bacteriocins from Lactobacillus sakei and Pediococcus spp.", abstract = "The present study consisted of an applied test in meat
system to assess the effectiveness of three bio agents bacteriocinproducing
strains: Lm24: Lactobacillus sakei, Lm14and Lm25:
Pediococcus spp. Two tests were carried out: The ex situ test was
intended for three batches added with crude bacteriocin solutions at
12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the
in situ one consisted of four batches; three of them inoculated with
one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one
was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The
two used tests were done in the presence of the pathogen
St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch
served as a positive or a negative control was used too. The
incubation was performed at 7°C. Total viable counts, staphylococci
and lactic acid bacteria, at the beginning and at selected times with
interval of three days were enumerated. Physico-chemical
determinations (except for in situ test): pH, dry mater, sugars, fat and
total protein, at the beginning and at end of the experiment, were
done, according to the international norms. Our results confirmed the
ex situ effectiveness. Furthermore, the batches affected negatively the
total microbial load over the incubation days, and showed a
significant regression in staphylococcal load at day seven, for Lm14,
Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed
that, at the last day of culture, staphylococcal load was nil for the
three batches. In the in situ test, the cultures displayed less inhibitory
attitude and recorded a decrease in staphylococcal load, for Lm14,
Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore,
physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an
increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry
mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result
reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%,
0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%,
5.37%, 5.5%. This study suggests that the use of selected strains as
Lm25 could lead to the best results and would help in preserving and
extending the shelf life of lamb meat.
", keywords = "Biocontrol, in situ and ex situ, meat system,
St. aureus, Lactobacillus sakei, Pediococcus spp.", volume = "8", number = "2", pages = "178-6", }
