Lactic acid bacteria capable of inhibiting campylobacter coli in vitro

ROMERO SCHARPEN, A.; FUHR, E.; BERISVIL, A.P.; SIGNORINI, M.L.; ZIMMERMANN, J.A.; BLAJMAN J.E.; FRIZZO, L.S.; ROSMINI, M.R.; ROMERO SCHARPEN, A.; FUHR, E.; SIGNORINI, M.L.; BLAJMAN J.E.; ROSMINI, M.R.; BERISVIL, A.P.; ZIMMERMANN, J.A.; FRIZZO, L.S.

Tucumán

Simposio; SIBAL 2016 V Simposio Internacional de Bacterias Lácticas; 2016

Centro de Referencia para Lactobacilos CERELA CONICET

Thermophilic Campylobacter(TC) are commensal microorganisms of birds, including poultry, and thesubspecie C. coli is one of the mostcommon bacterial causes of human intestinal diseases. The reduction of TC inpoultry would decrease the risk of infection for consumers. Probiotics,including lactic acid bacteria (LAB), can help to improve the intestinalmicrobial balance and therefore, decrease the capability of Campylobacter to colonize and grow inthe chicken caeca during primary production. The aim of this study was evaluatethe anti-Campylobacter coli activityof cell-free culture supernatant (CSF) of eleven LAB of avian origin with probioticscapacities. Inhibition activity was evaluated by the well diffusion test. Lactobacillus strains (L. salivarius DSPV001P, DSPV002P,DSPV003P, DSPV006P, DSPV007P, DSPV008P, DSPV010P y DSPV011P, L. agilis DSPV004P, DSPV005P y DSPV009P)were cultured in deMan, Rogosa and Sharpe (MRS) broth at 37ºC for 24 h.Lactobacillus cultures were harvested and the supernatants were divided in CSFneutralized (CSFn, adjustment the cell-free supernatant to pH of 7.0with 3N NaOH) and CSF unneutralized (CSFun). Suspensions of four C. coli isolated of chickens withdifferent pulse types obtained by PFGE (pulsed-field gel electrophoresis) wereused. C. coli were incubated innutritive agar for 48 h in a microaerophilic environment. The microbialsuspension with a concentration of 107 CFU/ml of Campylobacter was added to 15 ml ofnutritive agar (0,8% agar) and plated. Five mm-wells were punched on the agar, and35 µl of CSFn and CSFun of each strain of LAB was addedinto the holes. Lactic acid diluted ¼ was used as the positive control. Plateswere incubated for 48 h at 37ºC under microaerophilic conditions and thediameter of the inhibition zone around each well was measured. The trial wasreplicated three times. In all the cases the antagonic effect was observed bothCSFn and CSFun against four C. coli, which indicated that activity, was probably not exertedonly by pH. Six BAL showed halos of inhibition equal or higher to 15 mm againstat least 2 C. coli. Of these six,only L. salivarius DSPV006P and L. salivarius DSPV011P showed this behavior against the 4 C. coli. In conclusion, L. salivarius DSPV006P y L. salivarius DSPV011P showed the bestinhibition capabilities against growth of C.coli in this study.  Therefore, testswill be conducted to assess the presence the other inhibitory substances as theproduction of hydrogen peroxide and / or bacteriocins produced by LAB.