Effect of Lactobacillus casei subsp. casei R-68 Isolated from Dadih on the Procarcinogenic Enzyme Activity and Fecal Microflora Count of Rats Challenged with Pathogenic Bacteria

Usman Pato (1), Yusmarini Yusuf (2), Yudi Prasetia Nainggolan (3)
(1) Department of Agricultural Technology, Faculty of Agriculture, Universitas Riau, Pekanbaru, 28193, Riau, Indonesia
(2) Department of Agricultural Technology, Faculty of Agriculture, Universitas Riau, Pekanbaru, 28193, Riau, Indonesia
(3) Department of Agricultural Technology, Faculty of Agriculture, Universitas Riau, Pekanbaru, 28193, Riau, Indonesia
Fulltext View | Download
How to cite (IJASEIT) :
Pato, Usman, et al. “Effect of Lactobacillus Casei Subsp. Casei R-68 Isolated from Dadih on the Procarcinogenic Enzyme Activity and Fecal Microflora Count of Rats Challenged With Pathogenic Bacteria”. International Journal on Advanced Science, Engineering and Information Technology, vol. 9, no. 5, Oct. 2019, pp. 1656-62, doi:10.18517/ijaseit.9.5.8812.
The human digestive tract is a complex ecosystem that may contain bacteria, yeast, and other microflora, which have  harmful and beneficial effects on the host. Species of Lactobacillus and Bifidobacterium are most commonly used as probiotics. Lactobacillus casei subsp. casei R-68 (LCR-68) isolated from dadih, traditional fermented buffalo milk from West Sumatera has the potential to be used as probiotic. The purposes of the present study were to evaluate the ability of strain LCR68 to inhibit the growth of the pathogenic bacteria Listeria monocytogenes FNCC-0156 and Escherichia coli FNCC-19 and reduce the activity of fecal mutagen enzymes in Wistar rats. The in vivo test used 25 male Wistar rats with an average weight of 174 - 176 g. This study consisted of five groups of treatment with five rats of each group. The results show a significant increase in the growth in all groups, although  a significantly lower weight gain was observed in rats challenged with Listeria monocytogenes and fed fermented milk LCR-68. The counts of aerobic and anaerobic microbes were the same in all groups. Significantly higher counts of lactic acid bacteria were determined after the application of fermented milk LCR68. Significantly lower counts of Escherichia coli were also observed after the application of fermented milk LCR68. The presence of LCR-68 in fermented milk reduced the activity of β-glucuronidase and β-glucosidase significantly in the feces of Wistar rats. Therefore, the strain R-68 as a probiotic is expected to be able to prevent the formation of procarcinogenic compounds into carcinogens that cause cancer in the digestive tract.

W. H Holzapfel, P. Haberer, J. Snel, U. Schillinger and J. H. J. Huis in’t Veld, “Overview of Gut Flora And Probiotics”. Int. J. Food Microbiol, vol. 41, pp. 85-101, 1998.

FAO/WHO, “Guidelines for the Evaluation of Probiotics in Food”. Joint FAO/WHO Working Group Report on Drafting Guidelines for the Evaluation of Probiotics in Food. London, Ontario, Canada, 2002.

C. Hill, F. Guarner, G. Reid, G. R. Gibson, D. J. Merenstein, et al., “The International Scientific Association for Probiotics and Prebiotics Consensus Statement on the Scope and Appropriate Use of the Term Probiotic”, Nat. Rev. Gastroenterol. Hepatol, vol. 11, pp. 506-514, 2014.

Y. E. M. Dommels, R. A. Kemperman, Y. E. M. P. Zebregs, R. B. Draaisma, A. Jol, et al., “Survival of Lactobacillus reuteri DSM 17938 and Lactobacillus rhamnosus GG in the Human Gastrointestinal Tract with Daily Consumption of a Low-fat Probiotic Spread”, Appl. Environ. Microbiol., pp. 6198-6204, 2009.

E. Weichselbaum, Probiotics and Health: A Review of the Evidence. Nutrition Bulletins, vol. 34, pp. 340-373, 2009.

R. Fuller, “Probiotics in Man and Animals”, J. Appl. Bacteriol., vol. 66 (5), pp. 365-378, 1989.

C. Guerin-Danan, C. Chabanet, C. Pedone, F. Popot, P. Vaissade et al., “Milk Fermented with Yogurt Cultures and Lactobacillus casei Compared with Yogurt and Gelled Milk: Influence on Intestinal Microflora in Healthy Infants”, Am. J. Clin. Nutr., vol. 67, pp. 111-117,1998.

G. J. Jansen, A. C. M. Wildeboer-Veloo, D. van der Waaij and J. E. Degener, “Escherichia coli as a Probiotic?”, Infection, vol. 26 (4), pp. 232-233, 1998.

C. Gil-Turnes, A. F. dos Santos, F. W. da Cruz and A. V. Monteiro, “Properties of the Bacillus cereus Strain Used in Probiotic CenBiot”. Revista Microbiol., vol. 30, pp. 11-14, 1999.

L. H. Duc, H. A. Hong, T. M. Barbosa, A. O. Hendriques and S. M. Cutting, “Characterization of Bacillus Probiotics Available for Human Use”, Appl. Environ. Microbiol., vol. 70(4), pp. 2161-2171, 2004.

H. A. Hong, J. M. Huang, R. Khaneja, L. V. Hiep, M. C. Urdaci and S. M. Cutting, “The safety of Bacillus subtilis and Bacillus indicus as Food Probiotics”, J. Appl. Microbiol., vol. 105, pp. 510-520, 2008.

A. M. Shareef and A. S. A. Al-Dabbagh, “Effect of Probiotic (Saccharomyces cerevisiae) on Performance of Broiler Chicks”, Iraqi J. Vet. Sci., vol. 23, pp. 23-29, 2009.

K. M. R. Duarte, L. H. Gomes, A. C. K. Sampaio, J. Issakowicz, F. Rocha, T. P. Granato and S. R. Terra, “Saccharomyces cerevisiae Used as Probiotic: Strains Characterization and Cell Viability”, J. Agric. Vet. Sci., vol. 1(2), pp. 17-19, 2012.

A. Hosono, T. Tanabe and H. Otani, “Binding Properties of Lactic Acid Bacteria Isolated from Kefir Milk with Mutagenic Amino Acid Pyrolysates”, Milchwissenschaft, vol. 45, pp.647-651, 1990.

U.Pato, “Bile and Acid Tolerance Of Lactic Acid Bacteria Isolated from Dadih and their Antimutagenicity against Mutagenic Heated Tauco”, Asian-Australas. J. Anim. Sci., vol. 16, pp. 1680-1685, 2003.

I. S. Surono, U. Pato, Koesnandar and A. Hosono, “In vivo Antimutagenicity of Dadih Probiotic Bacteria towards Trp-P1”. Asian-Australas. J. Anim. Sci., vol. 22, pp. 119-123, 2009.

M. Gleeson, N. C. Bishop and L. Struszczak, ?Effects of Lactobacillus casei Shirota Ingestion on Common Cold Infection and Herpes Virus Antibodies in Endurance Athletes: A Placebo-Controlled, Randomized Trial”. J. Appl. Physiol., vol. 116, pp. 1555-1563, 2016.

C. M. Galdeano and G. Perdigon, “The Probiotic Bacterium Lactobacillus casei Induces Activation of the Gut Mucosal Immune System Through Innate Immunity”, Clin. Vaccine Immunol., vol. 13(2), pp. 219-226, 2006.

I. D. Kusumawati, E. Harmayani and W. Asmara, “Effect of Probiotic Lactobacillus sp. Dad13 on Humoral Immune Response of Balb/C Mice Infected with Salmonella typhimurium”, Indon. J. Biotechnol., vol. 11 (1), pp. 870-877, 2006.

A. N. Vlasova, S. Kandasary, K. S. Chattha, G. Rajashekara and L. J. Saif, “Comparison of Probiotic Lactobacilli and Bifidobacteria Effects, Immune Responses and Rotavirus Vaccines and Infection in Different Host Species”, Vet. Immunol Immunopathol., vol. 172, pp. 72-84, 2016.

A. J. McBain and G. T. Macfarlane, “Ecological and Physiological Studies on Large Intestinal Bacteria in Relation to Production of Hydrolytic aqnd Reductive Enzymes Involved in Formation of Genotoxic Metabolites”, J. Med. Microbiol.. vol. 47, pp. 407-416, 1998.

M. Mroczynska and Z. Libudzisz, ”β-Glucuronidase and β-glucosidase Activity of Lactobacillus and Enterococcus Isolated from Human Feces”, Pol. J. Microbiol., vol. 59, pp. 265-269, 2010.

O. Sreekumar and A. Hosono. “Immediate effect of Lactobacillus acidophilus on the Intestinal Flora and Fecal Enzymes of Rats and the in vitro inhibition of Escherichia coli in Coculture”, J. Dairy Sci., vol. 83, pp. 931-939, 2000.

L. J. Brady and D. D. Gallaher, “The Role of Probiotic Cultures in the Prevention of Colon Cancer”, J. Nutr., vol. 130, pp. 410-414, 2000.

G. Hawksworth, B. S. Draser and M. J. Hill, “Intestinal Bacteria And The Hydrolysis of Glycosidic Bonds”, J. Med. Microbiol., vol. 4, pp. 1445-1459, 1971.

C. B. Cole, R. Fuller, A.K. Mallet and I. R. Rowland, “The Influence of the Host on Expression of Intestinal Microbial Enzyme Activities Involved in Metabolism of Foreign Compounds”, J. Appl. Bacteriol., vol. 59, pp. 549-553, 1985.

B. R. Goldin, L. Swenson, J. Dwyer, M. Sexton and S. L. Gorbach, “Effect of Diet and Lactobacillusacidophilus Supplements on Human Fecal Bacterial Enzymes”, J. Nat. Cancer Inst., vol. 64, pp. 255-61, 1980.

B. R. Goldin and S. L. Gorbach, “The effect of Milk and Lactobacillus Feeding on Human Intestinal Bacterial Enzyme Activity”, Am. J. Cli. Nutr., vol. 39, pp. 756-761, 1984.

A. Hosono, R. Wardojo and H. Otani, “Microbial Flora in “Dadih”, a Traditional Fermented Milk in Indonesia”, Lebens. Wissen. Technol., vol. 22, pp. 20-24, 1989.

U. Pato and A. Hosono, “Hypocholesterolemic Effect of Indigenous Dadih Lactic Acid Bacteria by Deconjugation of Bile Salts”, Asian-Australas. J. Anim. Sci., vol. 17, pp. 1741-1745, 2004.

U. Pato, V. S. Djohan, F. Khairunnisa and R. D. H. Hasibuan, “Antibiotic Resistance and Antibacterial Activity of Dadih Originated Lactobacillus casei subsp. casei R-68 against Food Borne Pathogens”, Asian J. Microbiol. Biotech. Env. Sci., vol. 19, pp. 577-587, 2017.

O. V. Oyetayo, “Phenotypic Characterization and Assessment of the Inhibitory Potential of Lactobacillus Isolates from Different Sources”, Afric. J. Biotech., vol 3(7), pp. 355-357, 2004.

N. Xie, Y. Cui, Y. N. Yin, X. Zhao, J. W. Yang, et al., “Effects of Two Lactobacillus Strains on Lipid Metabolism and Intestinal Microflora in Rats Fed a High-Cholesterol Diet”, BMC Compl. Alter. Med., vol. 11, pp. 53, 2011.

J. Lee, Y. Kim, H. S. Yun, J. G. Kim, S. Oh and S. H. Kim, “Genetic and Proteomic Analysis of Factors Affecting Serum Cholesterol Reduction by Lactobacillus acidophilus A4”, Appl. Env. Microbiol., vol. 76(14), pp. 4829-4835, 2010.

S. R. Konstantinov, H. Smidt, D. L.A. Akkermans, L. Casini, P. Trevisi, et al., “Feeding of Lactobacillus sobrius Reduces Escherichia coli F4 Levels in the Gut and Promotes Growth of Infected Piglets”, FEMS Microbiol. Ecol., vol. 66, pp. 599-607, 2008.

R. Salaj, J. Å tofiloví¡, A. Å oltesoví¡, Z. Hertelyoví¡, E. Hijoví¡ et al., “The Effects of two Lactobacillus plantarum Strains on Rat Lipid Metabolism Receiving a High Fat Diet”, Sci. World J., vol. 5, pp. 1-7, 2013.

X. Bian, T. T. Wang, M. Xu, S.E. Evivie, G. W. Luo, H. Z. Liang, S. F. Yu, and G. C. Huo, “Effect of Lactobacillus Strains on Intestinal Microflora and Mucosa Immunity in Escherichia coli O157:H7-Induced Diarrhea in Mice”, Curr. Microbiol. ,DOI 10.1007/s00284-016-1010-3, 2016.

L. Strojny, A. Bomba, E. Hijova, A. Chmelarova, G. MojžiÅ¡ova et al., “Effects of a Probiotic In Combination With Prebiotics on Intestinal Lactobacilli and Coliforms and Activities of Bacterial Enzymes in 1,2-Dimethylhydrazine Exposed Rats”, Czech J. Anim. Sci., vol. 56, pp. 99-106, 2011.

P. Haberer, M. Du Toit, L. M. T. Dicks, F. Ahrens and W. H. Holzaptel, “Effect of Potentially Probiotic Lactobacilli on Faecal Enzyme Activity in Minipigs on a High-Fat, High-Cholesterol Diet - A Preliminary In vivo Trial”, Int. J. Food Microbiol., vol. 87, pp.287-291, 2003.

R. Waard, J. Garssen, G. C. A. M. Bokken and J. G. Vos, “Antagonistic activity of Lactobacillus casei Strain Shirota against Gastrointestinal Listeria monocytogenes Infection in Rats”. Int. J. Food. Microbiol., vol. 73, pp. 93-100, 2002.

L. Arul, G. Benita and P. Balasubramanian, “Functional Insight for β-glucuronidase in Escherichia coli and Staphylococcus sp. RLH1”, Bioinformation, vol. 2, pp. 339-343, 2008.

J. Nakamura, Y. Kubota, M. Miyaoka, T. Saitoh, F. Mizuno and Y. Benno, “Comparison of Four Microbial Enzymes In Clostridia And Bacteroides Isolated from Human Feces”, Microbiol. Immunol., vol. 46, pp. 487-490, 2002.

A. M. Stringer, R. J. Gibso, R. M. Logan, J. M. Bowen, A. S. Yeoh, et al., “Faecal Microflora And Beta-Glucuronidase Expression Are Altered in an Irinotecan Induced Diarrhea Model in Rats”, Cancer Biol. Ther., vol. 7, pp. 1919-1925, 2008.

I. Tryland and L. Fiksdal, “Enzyme Characteristics Of Beta-D-Galactosidase- and Beta-D-Glucuronidase-Positive Bacteria and their Interference in Rapid Methods for Detection of Waterborne Coliforms and Escherichia coli”, Appl. Env. Microbiol., vol. 64, pp. 1018-1023, 1998.

J. H. Chang, Y. Y. Shim, S. K. Cha, M. J. Reaney and K. M. Chee, “Effect of Lactobacillus acidophilus KFR1342 on the Development of Chemically-induced Precancerous Growths in the Rat Colon”, J. Med. Microbiol., vol. 61, pp.361-368, 2012.

E. Hijova, J. Kuzma, L. Strojny, A. Bomba, I. Bertkova, et al., “Effect of Lactobacillus plantarum lS/07 on Intestinal Bacterial Enzyme Activities in the Prevention of Cancer, Atherosclerosis and Dysbiosis”, Acta Vet. Beogr., vol. 66, pp.294-303, 2016.

C. B. Cole, R. Fuller and S. M. Carter, “Effect of Probiotic Supplements of Lactobacillus acidophilus and Bifidobacterium adolescentis 2204 on β-glucosidase and β-glucuronidase Activity in the Lower Gut of Rats Associated with a Human Faecal Flora”, Microbiol. Ecol. Health Dis., vol. 2, pp. 223-225, 1989.

A. Verma and G. Shukla, “Probiotics Lactobacillus rhamnosus GG, Lactobacillus acidophilus Suppresses DMH-induced pProcarcinogenic Fecal Enzymes and Preneoplastic Aberrant Crypt Foci in Early Colon Carcinogenesis in Sprague Dawley Rats”, Nut. Cancer, vol. 65, pp. 84-91, 2013.

D. K. Lee, S. Jang, E. H. Baek, M. J. Kim, K. S. Lee, H. S. Shin, M. J. Chung, J. E. Kim, K. O. Lee and N. J. Ha, “Lactic Acid Bacteria Affect Serum Cholesterol Levels, Harmful Fecal Enzyme Activity, and Fecal Water Content”, Lipids Health Dis., vol. 8, pp. 21-29, 2009.

Authors who publish with this journal agree to the following terms:

    1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
    2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
    3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).