H.M., K., W., A., S., M., Kotb, E. (2021). Exploring antimicrobial activity of Lactobacillus spp. (probiotics) isolated from raw cow’s milk against Staphylococcus aureus causing bovine mastitis. Journal of Applied Veterinary Sciences, 6(3), 60-69. doi: 10.21608/javs.2021.79310.1083
Kamel H.M.; Armanious W.; Morgan S.; Ebtsam E.Z. Kotb. "Exploring antimicrobial activity of Lactobacillus spp. (probiotics) isolated from raw cow’s milk against Staphylococcus aureus causing bovine mastitis". Journal of Applied Veterinary Sciences, 6, 3, 2021, 60-69. doi: 10.21608/javs.2021.79310.1083
H.M., K., W., A., S., M., Kotb, E. (2021). 'Exploring antimicrobial activity of Lactobacillus spp. (probiotics) isolated from raw cow’s milk against Staphylococcus aureus causing bovine mastitis', Journal of Applied Veterinary Sciences, 6(3), pp. 60-69. doi: 10.21608/javs.2021.79310.1083
H.M., K., W., A., S., M., Kotb, E. Exploring antimicrobial activity of Lactobacillus spp. (probiotics) isolated from raw cow’s milk against Staphylococcus aureus causing bovine mastitis. Journal of Applied Veterinary Sciences, 2021; 6(3): 60-69. doi: 10.21608/javs.2021.79310.1083
Exploring antimicrobial activity of Lactobacillus spp. (probiotics) isolated from raw cow’s milk against Staphylococcus aureus causing bovine mastitis
1Directorate of Veterinary Medicine, Giza Governorate, Ministry of Agriculture and Land Reclamation, Egypt
2Department of Microbiology, Faculty of veterinary medicine, Cairo University, Egypt
3Department of Food hygiene and control, Faculty of veterinary medicine, Cairo University, Egypt.
4Department of Udder Health and Neonatal Diseases, Animal Reproduction Research Institute, Egypt.
Receive Date: 05 June 2021,
Revise Date: 05 July 2021,
Accept Date: 09 July 2021
Abstract
Bovine mastitis is a costly disease in the dairy farms globally. The control of such disease is generally based on the prevention by the strict hygienic measures during milking. Other approaches include vaccination and the application of antibiotics. Regardless to these procedures, mastitis is not entirely under the control, thus increasing the need for alternative tactics. This study was conducted to isolate and identify lactic acid bacteria (LAB) from fresh cow’s milk which possess antibacterial activity that could be used for mastitis control. 146 isolates were recognized as (LAB) from105 milk tanks samples after being cultured anaerobically on de Man, Rogosa and Sharpe (MRS) agar plates for 48 hours at 37 oC and identified by general bacteriological investigation. Afterwards, 24 isolates were identified to belong to genus Lactobacillus using polymerase chain reaction (PCR), and for species level recognition MALDI-TOF MS (matrix-assisted laser desorption ionization-time of flight mass spectrometry) was used resulted in : L. fermentum (5), L. brevis (3), L. plantarum (4), L. paracasei (2), L. rhamnosus (3), L. pentosus (2), L. casei (3), L. raffinolactis (1) and L. mesenteroids (1). The antimicrobial activity of these strains against one of the major mastitis pathogens, S. aureus,was detected by the agar well diffusion assay and the modified double layer method , where L. casei, L. fermentum andL. plantarum possess the most inhibiting effect besides they have no hemolytic nor gelatin liquefaction activity when their safety profiles were evaluated. The result of the antibiotic susceptibility test revealed that these isolates were resistant to vancomycin (VA), neomycin (N) and gentamycin (CN). On the other hands, they were highly sensitive to amoxicillin clavulanic acid (AMC), levamisole (LEV), tetracycline (TE) and penicillin (P).The study suggests that L. casei, L. fermentum and L. plantarum are perfect candidates to be used as probiotics to help in preventing and controlling bovine mastitis caused by Staphylococcus aureus as they were proven to be safe and have antimicrobial activity againstthe organism.
BAUER, AW., KIRBY, WM., SHERRIS, JC. AND TURCK, M., 1966. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol.;45(4):493-496.
BEHNSEN J, DERIU E, SASSONE-CORSI M, AND RAFFATELLU M., 2013. Probiotics: Properties, Examples and Specific Applications. Cold Spring Harb Perspect Med.; 3: a010074. doi: 10.1101/cshperspect. a010074 PMID: 23457295.
CARBONNELLE, E., P. GROHS, H. JACQUIER, N. DAY, S. TENZA, A. DEWAILLY, O. VISSOUARN, M. ROTTMAN, J.-L. HERRMANN, I. PODGLAJEN AND RASKINE L., 2012. Robustness of two MALDI-TOF mass spectrometry systems for bacterial identification. J. Microbiol. Methods, 89: 133–136. http://dx.doi.org/10.1016/j.mimet. 2012.03.003.
CHALUPOVA, J., M. RAUS, M. SEDLAŘOVA AND ŠEBELA, M., 2014. Identification of fungal microorganisms by MALDI-TOF mass spectrometry. Biotechnol. Adv., 32: 230–241. http://dx. doi.org/10.1016/j.biotechadv. 2013.11.002.
COEURET, V., DUBERNET, S., BERNARDEAU, M., GUEGUEN, M. AND VERNOUX, J., 2003. Isolation, characterisation and identification of lactobacilli focusing mainly on cheeses and other dairy products. Le Lait, 83(4), 269-306. http://dx.doi.org/10.1051/lait:2003019.
COTTER, P. D., HILL, C. AND ROSS, R. P., 2005. Bacteriocins: developing innate immunity for food. Nat Rev Microbiol. 3(10):777±88. https://doi.org/10.1038/nrmicro1273PMID: 1620571.1
DEC, M., URBAN-CHMIEL, R., GNAT, S., PUCHALSKI, A. AND WERNICKI, A., 2014. Identification of Lactobacillus strains of goose origin using MALDI-TOF mass spectrometry and 16S-23S rDNA intergenic spacer PCR analysis. Research in Microbiology, 165(3), 190-201. http://dx.doi.org/10.1016/j.resmic.2014.02.003..
DOWARAH, R.; VERMA, A. AND AGARWAL, N., 2017. The Use of Lactobacillus as an Alternative of Antibiotic Growth Promoters in Pigs: A Review. Anim. Nutr. 2017, 3, 1–6.
DUBERNET, S., DESMASURES, N. AND GUEGUEN, M., 2002. A PCR-based method for identifcation of lactobacilli at the genus level. FEMS Microbiology Letters 214 (2002) 271-275.
DUŠKOVÁ, M., ŠEDO, O., KŠICOVÁ, K., ZDRÁHAL, Z. AND KARPÍŠKOVÁ, R., 2012. Identification of lactobacilli isolated from food by genotypic methods and MALDI-TOF MS. International Journal of Food Microbiology, 159(2), 107-114. http://dx.doi.org/10.1016/j.ijfoodmicro.2012.07.029.
ERCOLINI, D., MOSCHETTI, G., BLAIOTTA, G. AND COPPOLA, S., 2001. The potential of a polyphasic PCR dGGE approach in evaluating microbial diversity of natural whey cultures for water buffalo Mozzarella cheese production: bias of culture-dependent and culture independent analyses. Systematic and Applied Microbiology, 24(4), 610-617. http:// dx.doi.org/10.1078/0723-2020-00076.
ESPECHE MC, PELLEGRINO M, FROLA I, LARRIESTRA A, BOGNI C. AND NADER-MACIAS ME., 2012. Lactic acid bacteria from raw milk as potentially beneficial strains to prevent bovine mastitis. Anaerobe.; 18: 103–109. doi10.1016/j.anaerobe.01.002 PMID: 22261519
EVEN S, BOUCHARD D. AND LE LOIR Y., 2014. Lactic acid bacteria to modulate virulence expression in pathogenic bacteria: An alternative to killing? Interactive probiotics. CRC Press. Boca Raton, USA: Pessione E. (Eds.); pp. 52–80.
FAO/WHO., 2001. Health and Nutritional Properties of Probiotics in Food including Powder Milk with Live Lactic Acid Bacteria. Report of a Joint FAO/WHO Expert Consultation on Evaluation of Health and Nutritional Properties of Probiotics in food Including Powder Milk with Live Lactic Acid Bacteria. American Córdoba Park Hotel Córdoba, Argentina.
FULLER, R., 1989. Probiotics in man and animals. The Journal of Applied Bacteriology 66 (5): 365–378.
GEORGIEVA, R., YOCHEVA, L., TSEROVSKA, L., ZHELEZOVA, G., STEFANOVA, N., ATANASOVA, A., DANGULEVA, A., IVANOVA, G., KARAPETKOV, N., RUMYAN, N. AND KARAIVANOVA, E., 2015. Antimicrobial activity and antibiotic susceptibility of Lactobacillus and Bifidobacterium spp. intended for use as starter and probiotic cultures. Biotechnol Biotechnol Equip 29(1):84–91.
HALDER D, MANDAL S. .2015. Curd Lactobacilli with Probiotic Potentiality. Transl. Biomed., 6: (2-8),1-6. I HALDER, D.; MANDAL, M.; SHIV SEKHAR CHATTERJEE, NISHITH KUMAR PAL AND SHYAMAPADA MANDAL., 2017. Indigenous probiotic lactobacillus isolates presenting antibiotic like activity against human pathogenic bacteria. biomedicines, 5, 31; doi:10.3390/biomedicines5020031.
JACOBSEN, C.N., 1999. Screening of probiotic activities of forty-seven strain Lactobacillus ssp.by invitro techniques and evaluation of the colonization ability of five selected strains in umans. Appl. Environ. Microbial., 65: 4949-4956.
JOBBY, R., YASH FLORA, Y., BORA, A., JHA, P., KAWALKAR, H. AND DESAI, N., 2020. Exploring probiotic activity of lactobacillus sp. isolated from indigenous breeds of cattle milk and fecal samples in bhatan village, mh., in. current microbiology. 77:1184–1190c. doi.org/10.1007/s00284-020-01910-x.
KANAK, E. K. AND YILMAZ, S. O., 2018. Maldi-tof mass spectrometry for the identification and detection of antimicrobial activity of lactic acid bacteria isolated from local cheeses. Food Sci. Technol, Campinas, 39(Suppl. 2): 462-469. DOI: Dhttps://doi.org/10.1590/fst.19418.
KLAENHAMMER, T.R., KULLEN, M.J., 1999. Selection and design of probiotics, International journal of food microbiology, 50: 45-57.
LE MARECHAL, C, THIERY, R, VAUTOR, E. AND LE LOIR, Y., 2011. Mastitis impact on technological properties of milk and quality of milk products-a review. Dairy Sci Technol.; 91: 247–282.
LIASI, S. A., AZMI, T. I., HASSAN, M. D., SHUHAIMI, M., ROSFARIZAN, M. AND ARIFF, A. B., 2009. Antimicrobial activity and antibiotic sensitivity of three isolates of lactic acid bacteria from fermented fish product, Budu. Malaysian Journal of Microbiology, Vol 5(1), pp. 33-37.
LOURENS-HATTINGH, ANALIE, VILJOEN AND BENNIE, C., 2001. Yogurt as probiotic carrier food. International Dairy Journal. January 2001, Volume (11) 1-2: Pages 1–17.
MAMI A, HENNI JE. AND KIHAL M., 2008. Antimicrobial Activity of Lactobacillus species Isolated from Algerian Raw goat’s Milk against Staphylococcus aureus. World J. Dairy & Food Sci., 3 (2): 39-49.
MASSOL-DEYA, A.A., ODELSON, D.A., HICKEY, R.F. AND TIEDJE, J.M., 1995. Bacterial community fingerprinting of amplified 16S and 16–23S ribosomal DNA gene sequences and restriction endonuclease analysis (ARDRA). In: "MolecularMicrobial. Ecology Manual",3.3.2, pp. 289-296.
MOSTAFA, N. G., GEWAILY, E.M., EL-ZAMIK, F. I. AND ALI, A.S., 2019. Purification and specification of bacteriocin produced by some lactobacillus spp. isolated from food. Zagazig J. Agric. Res., 46: (2), 489-500.
NACEF, M., M. CHEVALIER, S. CHOLLET, D. DRIDER AND C. FLAHAUT, 2016. MALDI-TOF mass spectrometry for the identification of lactic acid bacteria isolated from a French cheese: The Maroilles. Int. J. Food Microbiol., 1-7.
NEMCOVÁ, R.,1997. Criteria for selection of lactobacilli for probiotic use. Vet Med (Praha). 1997 Jan;42(1):19-27. Slovak. PMID: 9123779
NOWROOZI, J. AND MIRZAII, M., 2004. Study of Lactobacillus as Probiotic bacteria. Irainian J. Publ. Hlth., 33(2): 1-7.
OLIVER, S., LEWIS, M., GILLESPIE, B., DOWLEN, H.H., JAENICKE, E. AND ROBERTS, R., 2004. Microbiological Procedures for the Diagnosis of Bovine Udder Infection and Determination of Milk Quality.https://www.semanticscholar.org/paper/Microbiological-Procedures-for-the-Diagnosis-of-and-Oliver-Lewis/f7d5c3ba0f4ac879698b51cf5288e51b1eccef94
ÖZCAN, N., EZIN, Ö., AKPOLAT, N., BOZDAĞ, H., METE, M. AND GÜL, K., 2016. Klinik örneklerde saptanan Candida türlerinin MALDI-TOF MS ile tiplendirilmesi. Dicle Medikal Journal, 43(3), 390-394.
REID, G., KIM, S. O. AND KOHLER, G. A., 2006. Selecting, testing and understanding probiotic microorganisms. FEMs Immunological Medicinal Microbiology, 46: 149-157.
RYAN, M.P., JACK, R.W., JOSTEN, M. AND JUNG, G., 1999. Extensive posttranslational Modification, Including Serine to D-Alanine Conversion, in the two-component lantibiotic, lacticin 3147. J. biol. Chem., 274: 37544-37550.
SENGUPTA, R, ALTERMANN, E., ANDERSON, RC., MCNABB, WC., MOUGHAN, PJ. AND ROY, NC., 2013. The role of cell surface architecture of lactobacilli in host-microbe interactions in the gastrointestinal tract. Mediators Inflame.; 2013: 237921. doi: 10.1155/2013/237921 PMID: 23576850 Clin. Infect. Dis. 2010; 50: 1551–1558. doi: 10.1086/652763 PMID: 20455694
SERIKBAYEVA, A., KONUSPAYEVA, G., FAYE, B., LOISEAU, G. AND NARMURATOVA, M., 2005. Probiotic properties of a sour-milk product: shubat from the camel milk. Desertification combat and food safety. IOS Press, p. 187-191.
SHIRLEY, J. C. AND JEAN, F., 2010 . Quick Tips About Using Probiotics with Wildlife in Rehabilitation. www.ewildagain.org
SHOKRYAZDAN, P; SIEO, C.C; KALAVATHY, R; LIANG, J.B; ALITHEEN, N.B; JAHROMI, M.F. AND AHMED, M., 2014. Probiotic potential of Lactobacillus strains with antimicrobial activity against some human pathogenic strains. Biomed. Res. Int., 2, 1
SOLEIMANI, N.A., KERMANSHAHI, R. K., YAKHCHALI, B., AND SATTARI, T. N., 2010. Antagonistic activity of probiotic lactobacilli against Staphylococcus aureus isolated from bovine mastitis. African Journal of Microbiology Research, 4(20), 2169-2173.
SULIEMAN, A.E.; OSAWA, R. AND TSENKOVA, R., 2007. Isolation and identification of lactobacilli from Garris, a Sudanese fermented camel's milk product.research Journal of Microbiology 2(2): 125.
TAMBEKAR, D.H., BHUTADA, S.A., CHOUDHARY, S.D. AND KHOND, M.D., 2009. Assessment of potential probiotic bacteria isolated from milk of domestic animals. J. Appl. Biosci. 15, 815–819.
TILLE, P. M., 2014. Bailey & Scott’s Diagnostic Microbiology (Thirteenth edition.). St. Louis, Missouri: Elsevier.
TOURET, T., OLIVEIRA, M. AND SEMEDO-LEMSADDEK, T., 2018. Putative probiotic lactic acid bacteria isolated from sauerkraut fermentations. PLoS ONE13(9): e0203501.
VANNIYASINGAM, J.; KAPILAN, R. AND VASANTHARUBA, S., 2019. Isolation and characterization of potential probiotic lactic acid bacteria isolated from cow milk and milk products.
VELOO, A.C.M., WELLING, G.W. AND DEGENER, J.E., 2011. The identification of anaerobic bacteria using MALDI-TOF MS. Anaerobe17: 211-212. Vet. Med., 42: 19-27.
VIECO-SAIZ, N.; BELGUESMIA, Y.; RASPOET, R., AUCLAIR, E.; GANCEL, F.; KEMPF, I. AND DRIDER, D., 2019. Benefits and Inputs from Lactic Acid Bacteria and Their Bacteriocins as Alternatives to Antibiotic Growth Promoters During Food- Animal Production. Front. Microbiol. 10, 57.
VITHANAGE, N. R., YEAGER, T. R., JADHAV, S. R., PALOMBO, E. A., AND DATTA, N., 2014. Comparison of identification systems for psychrotrophic bacteria isolated from raw bovine milk. International Journal of Food Microbiology, 189, 26-38. doi.org/10.1016/j. ijfoodmicro.2014.07.023. PMid:25113043.
VLKOVA, E., RADA, V., POPELÁŘOVÁ, P., TROJANOVÁ, I. AND KILLER, J., 2006. Antimicrobial susceptibility of bifidobacteria isolated from gastrointestinal tract of calves. Livestock Science 105, 253.
WALLIS JK, KROMKER V. AND PADUCH JH., 2018. Biofilm formation and adhesion to bovine udder epithelium of potentially probiotic lactic acid bacteria. AIMS Microbiol. 4(2):209–224.