Antagonistic activity of lactic acid bacteria isolated from Minas artisanal cheeses against Brucella abortus

This study aimed to evaluate methods for studying the in vitro antimicrobial activity of lactic acid bacteria (LAB) against Brucella abortus and to evaluate the antagonistic effect of LAB on the viability of this pathogen. A total of 18 LAB strains (Lactobacillus plantarum, n = 11; Pediococcus acidilactici, n = 1; Lactobacillus rhamnosus, n = 4; and Lactobacillus brevis, n = 2), isolated from Minas artisanal cheeses produced in three regions (Canastra, Campos das Vertentes, and Araxá) of Minas Gerais State, Brazil, were tested for their antimicrobial activity against B. abortus using three methods: spot-on-lawn, agar well diffusion assay, and antagonistic activity of the culture supernatants. None of the tested LAB strains could inhibit B. abortus in the spot-on-lawn and agar-well diffusion assays. The supernatants produced by LAB had an acidic pH, with intensity depending on bacterial growth and strain, and could inhibit the growth of B. abortus. In contrast, pH-neutralized (pH 7.0) LAB supernatants did not suppress the growth of B. abortus. The results showed that the best technique to study the in vitro antagonism of LAB against B. abortus was the antagonistic activity of culture supernatants. The growth of B. abortus may have been inhibited by acid production.

[1]  A. Terpou,et al.  Assessment of the probiotic potential of lactic acid bacteria isolated from kefir grains: evaluation of adhesion and antiproliferative properties in in vitro experimental systems , 2019, Annals of Microbiology.

[2]  J. Nicoli,et al.  In vitro assessment of the probiotic potential of lactobacilli isolated from Minas artisanal cheese produced in the Araxá region, Minas Gerais state, Brazil , 2019, Arquivo Brasileiro de Medicina Veterinária e Zootecnia.

[3]  Maryam Dadar,et al.  Human brucellosis caused by raw dairy products: A review on the occurrence, major risk factors and prevention. , 2019, International journal of food microbiology.

[4]  Jamili Maria Suhet Mussi Efeito do antagonismo in vitro de bactérias ácido-láticas e da maturação na sobrevivência de brucella abortus em queijos tipo Minas artesanal , 2018 .

[5]  J. Nicoli,et al.  Assessment of the probiotic potential of lactic acid bacteria isolated from Minas artisanal cheese produced in the Campo das Vertentes region, Brazil , 2017 .

[6]  E. Dorneles,et al.  Molecular epidemiology of Brucella abortus isolated from cattle in Brazil, 2009-2013. , 2017, Acta tropica.

[7]  M. Souza,et al.  Isolation and identification of lactic acid bacteria from Brazilian Minas artisanal cheese , 2016 .

[8]  M. Heinemann,et al.  Analysis of 15 years of the National Program for the Control and Eradication of Animal Brucellosis and Tuberculosis, Brazil , 2016 .

[9]  N. J. Pimentel-Filho,et al.  Determining the minimum ripening time of artisanal Minas cheese, a traditional Brazilian cheese , 2015, Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology].

[10]  Gilson de Assis Sales Caracterização microbiológia e físico-química de queijo Minas artesanal da microrregião de Araxá-MG durante a maturação em diferentes épocas do ano , 2015 .

[11]  F. P. Poester,et al.  Brucella abortus S19 and RB51 vaccine immunogenicity test: Evaluation of three mice (BALB/c, Swiss and CD-1) and two challenge strains (544 and 2308). , 2015, Vaccine.

[12]  M. Elschner,et al.  Detection of Brucella melitensis in bovine milk and milk products from apparently healthy animals in Egypt by real-time PCR. , 2014, Journal of infection in developing countries.

[13]  C.F.A.M. Penna,et al.  Propriedades probióticas in vitro de Lactobacillus spp. isolados de queijos minas artesanais da Serra da Canastra - MG , 2014 .

[14]  J. P. Mol,et al.  Genetic stability of Brucella abortus isolates from an outbreak by multiple-locus variable-number tandem repeat analysis (MLVA16) , 2014, BMC Microbiology.

[15]  M. Souza,et al.  Potencial probiótico in vitro de bactérias ácido-láticas isoladas de queijo-de-minas artesanal da Serra da Canastra, MG , 2013 .

[16]  Orla O'Sullivan,et al.  The complex microbiota of raw milk. , 2013, FEMS microbiology reviews.

[17]  E. D. De Martinis,et al.  Identification and evaluation of the probiotic potential of Lactobacillus paraplantarum FT259, a bacteriocinogenic strain isolated from Brazilian semi-hard artisanal cheese. , 2013, Anaerobe.

[18]  S. Esposito,et al.  Detection of Brucella spp. in Stretched Curd Cheese as Assessed by Molecular Assays , 2013 .

[19]  D. Pfeiffer,et al.  A "One Health" surveillance and control of brucellosis in developing countries: moving away from improvisation. , 2013, Comparative immunology, microbiology and infectious diseases.

[20]  T. Phister,et al.  Microbial interactions in food fermentations. , 2013, Annual review of food science and technology.

[21]  S. Fillmore,et al.  Antimicrobial activity of bacteriocin-producing lactic acid bacteria isolated from cheeses and yogurts , 2012, AMB Express.

[22]  M. Kihal,et al.  Antimicrobial activity of lactic acid bacteria and the spectrum of their biopeptides against spoiling germs in foods , 2012 .

[23]  S. Menezes QUEIJO ARTESANAL: IDENTIDADE, PRÁTICA CULTURAL E ESTRATÉGIA DE REPRODUÇÃO SOCIAL EM PAÍSES DA AMÉRICA LATINA , 2012 .

[24]  B. Dal Bello,et al.  Technological characterization of bacteriocin producing Lactococcus lactis strains employed to control Listeria monocytogenes in cottage cheese. , 2012, International journal of food microbiology.

[25]  C.F.A.M. Penna,et al.  Queijo de minas artesanal da Serra da Canastra: influência da altitude das queijarias nas populações de bactérias acidolácticas , 2011 .

[26]  A. López-Merino,et al.  Brucella melitensis survival during manufacture of ripened goat cheese at two temperatures. , 2011, Foodborne pathogens and disease.

[27]  L. Perin,et al.  Protocols for the isolation and detection of lactic acid bacteria with bacteriocinogenic potential , 2010 .

[28]  P. Monsan,et al.  In vitro screening of probiotic lactic acid bacteria and prebiotic glucooligosaccharides to select effective synbiotics. , 2010, Anaerobe.

[29]  N. Benomar,et al.  Microbial antagonists to food-borne pathogens and biocontrol. , 2010, Current opinion in biotechnology.

[30]  R. Dias,et al.  Detection of Brucella abortus DNA in illegal cheese from São Paulo and Minas Gerais and differentiation of B19 vaccinal strain by means of the polymerase chain reaction (PCR) , 2007 .

[31]  S. Olsen,et al.  Efficacy of strain RB51 vaccine in heifers against experimental brucellosis. , 2006, Vaccine.

[32]  P. Hütt,et al.  Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero‐ and uropathogens , 2006, Journal of applied microbiology.

[33]  L. Dicks,et al.  Characterization of a 3944 Da bacteriocin, produced by Enterococcus mundtii ST15, with activity against Gram-positive and Gram-negative bacteria. , 2005, International journal of food microbiology.

[34]  A. Halasz,et al.  Influence of Growth Medium on Hydrogen Peroxide and Bacteriocin Production of Lactobacillus Strains , 2005 .

[35]  M. Mataragas,et al.  Influence of nutrients on growth and bacteriocin production by Leuconostoc mesenteroides L124 and Lactobacillus curvatus L442 , 2004, Antonie van Leeuwenhoek.

[36]  A. Lage,et al.  Brucellosis in Brazil. , 2002, Veterinary microbiology.

[37]  R. P. Ross,et al.  Preservation and fermentation: past, present and future. , 2002, International journal of food microbiology.

[38]  J. Swings,et al.  Influence of the culture medium on antibiotic susceptibility testing of food‐associated lactic acid bacteria with the agar overlay disc diffusion method , 2002, Letters in applied microbiology.

[39]  M. Hamdy,et al.  Detection of Brucella species in the milk of infected cattle, sheep, goats and camels by PCR. , 2002, Veterinary journal.

[40]  P. McSweeney,et al.  Advances in the study of proteolysis during cheese ripening , 2001 .

[41]  J. Rodríguez-Otero,et al.  Microbiological, chemical and biochemical characteristics of ‘Tetilla’ raw cows-milk cheese , 2001 .

[42]  I. Rogelj,et al.  Bacteriocin complex of Lactobacillus acidophilus LF221 — production studies in MRS media at different pH values and effect against Lactobacillus helveticus ATCC 15009 , 1998 .

[43]  T. Nawas,et al.  The effect of the pH of various dairy products on the survival and growth of Brucella melitensis. , 1990, Annals of tropical medicine and parasitology.

[44]  M. Daeschel,et al.  Antimicrobial Activity of Lactic Acid Bacteria Against Listeria monocytogene s. , 1989, Journal of food protection.

[45]  H L Houtzager,et al.  Antonie van Leeuwenhoek. , 1983, European journal of obstetrics, gynecology, and reproductive biology.

[46]  J. Tagg,et al.  Copyright � 1995, American Society for Microbiology Bacteriocins of Gram-Positive Bacteria , 1995 .

[47]  G. Davies,et al.  The survival of Brucella abortus in milk and milk products. , 1973, The British veterinary journal.

[48]  C. Kuzdas,et al.  The survival of Brucella abortus, U.S.D.A. strain 2308, under controlled conditions in nature. , 1954, The Cornell veterinarian.

[49]  J. O. Irwin,et al.  The estimation of the bactericidal power of the blood , 1938, Epidemiology and Infection.

[50]  C. L. D. Paula,et al.  Detecção de Brucella spp. em leite bovino não pasteurizado através da Reação de Cadeia pela Polimerase (PCR) , 2015 .

[51]  R. Kara,et al.  Investigation of Brucella abortus and Brucella melitensis at Cheeses in Afyonkarahisar, Turkey , 2013 .

[52]  M. Corbel,et al.  Brucellosis in humans and animals. , 2006 .

[53]  Eva María Santos López,et al.  Survival of Brucella abortus in milk fermented with a yoghurt starter culture. , 2005, Revista latinoamericana de microbiologia.

[54]  T. Beresford,et al.  The Microbiology of Cheese Ripening , 2004 .