Skip to main content
SpringerLink
Log in

In Vitro Evaluation of the Probiotic and Safety Properties of Bacteriocinogenic and Non-Bacteriocinogenic Lactic Acid Bacteria from the Intestines of Nile Tilapia and Common Carp for Their Use as Probiotics in Aquaculture

  • Published:
Probiotics and Antimicrobial Proteins Aims and scope Submit manuscript

Abstract

In this study, seven bacteriocinogenic and non-bacteriocinogenic LAB strains previously isolated from the intestines of Nile tilapia and common carp and that showed potent antibacterial activity against host-derived and non-host-derived fish pathogens were assayed for their probiotic and safety properties so as to select promising candidates for in vivo application as probiotic in aquaculture. All the strains were investigated for acid and bile tolerances, transit tolerance in simulated gastrointestinal conditions, for cell surface characteristics including hydrophobicity, co-aggregation and auto-aggregation, and for bile salt hydrolase activity. Moreover, haemolytic, gelatinase and biogenic amine-producing abilities were investigated for safety assessment. The strains were found to be tolerant at low pH (two strains at pH 2.0 and all the strains at pH 3.0). All of them could also survive in the presence of bile salts (0.3% oxgall) and in simulated gastric and intestinal juices conditions. Besides, three of them were found to harbour the gtf gene involved in pH and bile salt survival. The strains also showed remarkable cell surface characteristics, and 57.14% exhibited the ability to deconjugate bile salts. When assayed for their safety properties, the strains prove to be free from haemolytic activity, gelatinase activity and they could neither produce biogenic amines nor harbour the hdc gene. They did not also show antibiotic resistance, thus confirming to be safe for application as probiotics. Among them, Lactobacillus brevis 1BT and Lactobacillus plantarum 1KMT exhibited the best probiotic potentials, making them the most promising candidates.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  1. FAO (Food and Agriculture Organization of the United Nations) (2016) The state of world fisheries and aquaculture—contributing to food security and nutrition for all. FAO fisheries and aquaculture technical paper, Rome, p 204

    Google Scholar 

  2. Sahoo TK, Jena PK, Nagar N, Patel AK, Seshadri S (2015) In Vitro evaluation of probiotic properties of lactic acid bacteria from the gut of Labeo rohita and Catla catla. Probiotics Antimicrob Proteins. doi:10.1007/s12602-015-9184-8

  3. Tekwombuo J (2013) Hatchery design and brood stock management policy as a tool for sustainable aquaculture: case of Cameroon. United Nations University Fisheries Training Programme, Iceland http://www.unuftp.is/static/fellows/dociment/joseph13prf.pdf

    Google Scholar 

  4. Kaktcham PM, Zambou NF, Fonteh AF, Pérez-Chabela ML (2015) Aquaculture in Cameroon and potential of lactic acid bacteria to be used as diseases controlling agents. A review. Nacameh 9(1):1–18

    Google Scholar 

  5. Kaktcham PM, Temgoua J-B, Zambou NF, Diaz-Ruiz G, Wacher C, Perez-Chabela ML (2017) Quantitative analyses of the bacterial microbiota of rearing environment, tilapia and common carp cultured in earthen ponds and inhibitory activity of its lactic acid bacteria on fish spoilage and pathogenic bacteria. World J Microbiol Biotechnol 33(2):1–12

    Article  Google Scholar 

  6. Martínez Cruz P, Ibáñez AL, Monroy Hermosillo OA, Ramírez Saad HC (2012) Use of probiotics in Aquaculture. Int Sch Res Netw 2012:916845 13p

    Google Scholar 

  7. Giri SS, Sukumaran V, Sen SS, Vinumonia J, Nazeema B, Jena PK (2011) Antagonistic activity of cellular components of probiotic bacteria, isolated from the gut of Labeo rohita, against Aeromonas hydrophila. Probiotics Antimicrob Proteins 3:214–222

    Article  CAS  Google Scholar 

  8. Sala BG (2013) Isolation, identification and antimicrobial activity of bacteriocinogenic lactic acid bacteria from marine origin: biochemical and genetic characterisation and heterologous production of curvacins G14 and G15 from Lactobacillus curvatus BCS35. Ph.D thesis, University Complutense of Madrid, pp 177–224

  9. Verschuere L, Rombaut G, Sorgeloos P, Verstraete W (2000) Probiotic bacteria as biological control agents in aquaculture. Microbiol Mol Biol Rev 64:655–671

    Article  CAS  Google Scholar 

  10. Austin B, Austin DA (2007) Bacterial fish pathogens: diseases of farmed and wild fish, 4th edn. Springer-Praxis, Chichester

    Google Scholar 

  11. Gatesoupe FJ (2008) Updating the importance of lactic acid bacteria in fish farming: natural occurrence and probiotic treatments. J Mol Microbiol Biotechnol 14:107–114

    Article  CAS  Google Scholar 

  12. Giri SS, Sukumaran V, Dangi NK (2012) Characteristics of bacterial isolates from the gut of freshwater fish, Labeo rohita that may be useful as potential probiotic bacteria. Probiotics Antimicrob Proteins 4:238–242

    Article  Google Scholar 

  13. Muñoz-Atienza E, Gómez-Sala B, Araújo C, Campanero C, del Campo R, Hernández PE, Herranz C, Cintas LM (2013) Antimicrobial activity, antibiotic susceptibility and virulence factors of lactic acid bacteria of aquatic origin intended for use as probiotics in aquaculture. BMC Microbiol 13:15

    Article  Google Scholar 

  14. Balcázar JL, Blas I, Ruiz-Zarzuela I, Cunningham D, Vendrell D, Muzquiz JL (2006) The role of probiotics in aquaculture. Vet Microbiol 114:173–186

    Article  Google Scholar 

  15. Kumar R, Mukherjee SC, Paniprasad K, Pal AK (2006) Evaluation of Bacillus subtilis as a probiotic to Indian major carp Labeo rohita (ham.) Aquacult Res 37:1215–1221

    Article  Google Scholar 

  16. Son VM, Chang CC, Wu MC, Guu YK, Chiu CH, Cheng W (2009) Dietary administration of the probiotic, Lactobacillus plantarum, enhanced the growth, innate immune responses, and disease resistance of the grouper Epinephelus coioides. Fish Shellfish Immunol 26:691–698

    Article  CAS  Google Scholar 

  17. Verdenelli MC, Ghelfi F, Silvi S, Orpianesi C, Cecchini C, Cresci A (2009) Probiotic properties of Lactobacillus rhamnosus and Lactobacillus paracasei isolated from human faeces. Eur J Nutr. doi:10.1007/s00394-009-0021-2

  18. Stack HM, Kearney N, Stanton C, Fitzgerald GF, Ross RP (2010) Association of betaglucan endogenous production with increased stress tolerance of intestinal lactobacilli. Appl Environ Microbiol 76:500–507

    Article  CAS  Google Scholar 

  19. Vrancken G, Rimaux T, Weckx S, De Vuyst L, Leroy F (2009) Environmental pH determines citrulline and ornithine release through the arginine deiminase pathway in Lactobacillus fermentum IMDO 130101. Int J Food Microbiol 135:216–222

    Article  CAS  Google Scholar 

  20. Corcoran BM, Stanton C, Fitzgerald GF, Ross RP (2005) Survival of probiotic lactobacilli in acidic environments is enhanced in the presence of metabolizable sugars. Appl Environ Microbiol 71(6):3060–3067

    Article  CAS  Google Scholar 

  21. Dashkevicz MP, Feighner SD (1989) Development of a differential medium for bile salt hydrolase-active Lactobacillus spp. Appl Environ Microbiol 55:11–16

    CAS  Google Scholar 

  22. Ekmekci H, Aslim B, Ozturk S (2009) Characterization of vaginal lactobacilli coaggregation ability with Escherichia coli. Microbiol Immunol 53:59–65

    Article  CAS  Google Scholar 

  23. ISO 10932/IDF 223 (2010) Milk and milk products—determination of minimal inhibitory concentration (MIC) of antibiotics applicable to bifidobacteria and non-enterococcal lactic acid bacteria (LAB)

  24. EFSA (2012) Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J 10(6):2740

    Google Scholar 

  25. Danielsen M, Wind AA (2003) Susceptibility of Lactobacillus ssp. to antimicrobial agents. Int J Food Microbiol 82:1–11

    Article  CAS  Google Scholar 

  26. CLSI (Clinical and Laboratory Standards Institute) (2009) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; Approved standard-Eight Edition. CLSI document M07-A8 (ISBN 1–56238–689-1). Clinical and Laboratory Standards Institute, 940 West Valley Road, suite 1400, Wayne, Pennsylvania 19087–1898 USA

  27. Gerhardt P, Murray RG, Costilow RN, Nester EW, Wood WA, Krieg NR, Phillips GB (1981) Manual of methos for general bacteriology. American Soc Microbiol. Washington, DC 20006

  28. Harrigan WF, Mc Cance ME (1990) Laboratory methods. In: Harigan WF, Mc Cance ME (eds) Food and dairy microbiology. Academic press, New York, pp 12–15

    Google Scholar 

  29. Bover-Cid S, Holzapfel WH (1999) Improved screening procedure of biogenic amine production by lactic acid bacteria. Int J Food Microbiol 53:33–41

    Article  CAS  Google Scholar 

  30. Costantini A, Cersosimo M, Del Prete V, Garcia-Moruno E (2006) Production of biogenic amines by lactic acid bacteria: screening by PCR thin-layer chromatography, and high-performance liquid chromatography of strains isolated from wine and must. J Food Prot 69:391–396

    Article  CAS  Google Scholar 

  31. Buntin N, Chanthachum S, Hongpattarakere T (2008) Screening of lactic acid bacteria from gastrointestinal tracts of marine fish for their potential use as probiotics. Songklanakarin J Sci Technol 30(Suppl.1):141–148

    Google Scholar 

  32. Balcázar JL, Vendrell D, Blas ID, Ruiz-Zarzuela I, Muzquiz JL, Girones O (2008) Characterization of probiotic properties of lactic acid bacteria isolated from intestinal microbiota of fish. Aquaculture 278:188–191

    Article  Google Scholar 

  33. Chemlal-Kherraz D, Sahnouni F, Matallah-Boutiba A, Boutiba Z (2012) The probiotic potential of lactobacilli isolated from Nile tilapia (Oreochromis niloticus)’s intestine. Afr J Biotechnol 11(68):13220–13227

    Google Scholar 

  34. Turpin W, Humblot C, Guyot JP (2011) Genetic screening of functional properties of lactic acid bacteria in a fermented pearl millet slurry and in the metagenome of fermented starchy foods. Appl Environ Microbiol 77(24):8722–8734

    Article  CAS  Google Scholar 

  35. Vizoso Pinto MG, Franz CMAP, Schillinger U, Holzapfel WH (2006) Lactobacillus spp. with in vitro probiotic properties from human faeces and traditional fermented products. Int J Food Microbiol 109:205–214

    Article  CAS  Google Scholar 

  36. Argyri AA, Zoumpopoulou G, Karatzas K-AG, Tsakalidou E, Nychas G-JE, Panagou EZ, Tassou CC (2013) Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests. Food Microbiol 33:282–291

    Article  CAS  Google Scholar 

  37. Solieri L, Bianchi A, Mottolese G, Lemmetti F, Giudici P (2014) Tailoring the probiotic potential of non-starter Lactobacillus strains from ripened Parmigiano Reggiano cheese by in vitro screening and principal component analysis. Food Microbiol 38:240–249

    Article  CAS  Google Scholar 

  38. González-Vázquez R, Azaola-Espinosa A, Mayorga-Reyes L, Reyes-Nava LA, Shah NP, Rivera-Espinoza Y (2015) Isolation, identification and partial characterization of a Lactobacillus casei strain with bile salt hydrolase activity from pulque. Probiotics Antimicrob Proteins 7(4):242–248

    Article  Google Scholar 

  39. Todorov SD, Holzapfel W, Nero LA (2016) In Vitro evaluation of beneficial properties of bacteriocinogenic Lactobacillus plantarum ST8Sh. Probiotics Antimicrob Proteins 9:194–203

    Article  Google Scholar 

  40. Kos B, Suskovic J, Vukovic S, Simpraga M, Frece J, Matosic S (2003) Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 94:981–987

    Article  CAS  Google Scholar 

  41. Kaktcham PM, Zambou NF, Tchouanguep MF, El-Soda M, Iqbal Choudhary M (2012) Antimicrobial and safety properties of Lactobacillus strains isolated from two Cameroonian traditional fermented foods. Sci Pharm 80(1):189–203

    Article  CAS  Google Scholar 

  42. Elkins CA, Mullis LB (2004) Bile-mediated aminoglycoside sensibility in Lactobacillus species likely results from increased membrane permeability attributable to cholic acid. Appl Environ Microbiol 70:7200–7209

    Article  CAS  Google Scholar 

  43. Herreros MA, Fresno JM, Sandoval H, Gonzáles L, Castro JM, Tornadijo ME (2004) Antimicrobial activity and antibiotic resistance of lactic acid bacteria isolated from Armada cheese (a Spanish goats’ milk cheese). Food Microbiol 22:455–459

    Article  Google Scholar 

  44. Rojo-Bezares B, Sáenz Y, Poeta P, Zarazaga M, Ruiz-Larrea F, Torres C (2006) Assessment of antibiotic susceptibility within lactic acid bacteria strains isolated from wine. Int J Food Microbiol 111:234–240

    Article  CAS  Google Scholar 

  45. Kumar A, Kumar D (2014) Isolation and characterization of bacteria from dairy samples of Solan in Himachal Pradesh for identification of Lactobacillus spp. Int J Pharm Sci Rev Res 25:110–114

    Google Scholar 

  46. Charteris WP, Kelly PM, Morelli L, Collins JK (2001) Gradient diffusion antibiotic susceptibility testing of potentially probiotic lactobacilli. J Food Prot 64:2007–2014

    Article  CAS  Google Scholar 

  47. Katla AK, Kruse H, Johnsen G, Herikstad H (2001) Antimicrobial susceptibility of starter culture bacteria used in Norwegian dairy products. Int J Food Microbiol 67:147–152

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to TWAS (The World Academy of Sciences for the development of science in developing countries) and CONACyT (Consejo Nacional de Ciencia y Tecnología, México) for their support through TWAS-CONACyT postdoctoral fellowship (FR number: 3240275624) awarded to K.P.M.

Author information

Authors and Affiliations

  1. Laboratory of Biochemistry, Food Science and Nutrition (LABPMAN), Department of Biochemistry, Faculty of Science, University of Dschang, Cameroon, P.O Box 67, Dschang, Cameroon

    Pierre Marie Kaktcham, Jules-Bocamdé Temgoua & François Ngoufack Zambou

  2. Departamento de Biotecnología, Universidad Autónoma Metropolitana-Iztapalapa (UAM-I), Av. San Rafael Atlixco 186, 09340, Mexico, Distrito Federal, Mexico

    Pierre Marie Kaktcham & María de Lourdes Pérez-Chabela

  3. Departamento de Alimentos y Biotecnología, Facultad de Química, Universidad Nacional Autónoma de México (UNAM), Ciudad Universitaria Coyoacán, 04510, Mexico, Distrito Federal, Mexico

    Gloria Diaz-Ruiz & Carmen Wacher

Authors
  1. Pierre Marie Kaktcham
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jules-Bocamdé Temgoua
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. François Ngoufack Zambou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gloria Diaz-Ruiz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carmen Wacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. María de Lourdes Pérez-Chabela
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pierre Marie Kaktcham.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Research Involving Human Participants and/or Animals

Not applicable.

Informed Consent

Not applicable.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kaktcham, P.M., Temgoua, JB., Zambou, F.N. et al. In Vitro Evaluation of the Probiotic and Safety Properties of Bacteriocinogenic and Non-Bacteriocinogenic Lactic Acid Bacteria from the Intestines of Nile Tilapia and Common Carp for Their Use as Probiotics in Aquaculture. Probiotics & Antimicro. Prot. 10, 98–109 (2018). https://doi.org/10.1007/s12602-017-9312-8

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12602-017-9312-8

Keywords

Search

Navigation