Isolation of bacteriocinogenic lactic acid bacteria and their application in the control of Listeria monocytogenes in fresh goat cheese

Author(s)

Danielle Nader Furtado

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ)
• Defense date: 2010-03-04

Examining board members

Bernadette Dora Gombossy de Melo Franco; Alcina Maria Liserre; Elaine Cristina Pereira de Martinis

• Advisor: Bernadette Dora Gombossy de Melo Franco

Abstract

Listeria monocytogenes causes listeriosis, a serious zoonotic disease that causes infections in the central nervous system (meningitis, encephalitis and meningoencephalitis), bacteremia and septicemia. The disease presents low morbidity and high mortality and affects mainly those in the risk group, such as pregnant women, neonates, immunocompromised individuals and the elderly. L. monocytogenes has been frequently detected in in natura and processed foods. Like cheeses and other dairy products. This study aimed to isolate lactic acid bacteria capable of producing antimicrobial compounds from goat milk, identify the isolates, characterize the bacteriocins and evaluate their potential application in controlling the growth of L. monocytogenes in goat cheese during storage at 8-10°C. Six bacteriocinogenic strains (DF2Mi, DF3Mi, DF4Mi, DF5Mi, DF6Mi e DF60Mi) were successfully isolated from raw goat milk. Using appropriate phenotypic tests and 16S rRNA sequencing, these strains were identified as Lactococcus lactis subsp. lactis (DF2Mi, DF3Mi, DF4Mi e DF5Mi), Leuconostoc lactis (DF6Mi) and Lactobacillus paracasei subsp. paracasei (DF60Mi). The physico-chemical and biological characterization of the bacteriocins produced by the strains DF4Mi, DF6Mi e DF60Mi indicated that they were resistant to heat and pH extremes, but presented different spectrum of activity, sensitivity to chemicals, adsorption to target cells and lysis of L. monocytogenes. The effect of pH, temperature and culture media composition in bacteriocin production was also strain-dependent. The strain DF4Mi presented the best antimicrobial activity and was selected for the studies on inhibition of L. monocytogenes in cheese. Frescal cheese was manufactured with pasteurized goat milk added of a culture of DF4Mi (106 CFU/mL), and experimentally contaminated with L. monocytogenes (103 CFU/g). Control cheeses were also prepared: those added of 12.5 mg/Kg nisin (positive control) and those added of a non-bacteriocinogenic L. lactis subsp. lactis strain. The strain presented a bacteriostatic effect, controlling the growth of the pathogen for 10 days at 8-10°C. However, a similar effect was observed in the cheeses prepared with the non-bacteriocinogenic strain, indicating that the inhibition cannot be credited to bacteriocins. In the cheeses containing nisin, a 2 log reduction in the counts of L. monocytogenes was achieved after 10 days at 8-10°C. In the cheeses with no added nisin or lactic acid bacteria, the counts of L. monocytogenes after 10 days at 8-10°C were high (106 CFU/g). (AU)