Use of probiotics to reduce faecal shedding of Shiga toxin-producing Escherichia coli in sheep.

Shiga toxin-producing Escherichia coli (STEC) are zoonotic, foodborne pathogens of humans. Ruminants, including sheep, are the primary reservoirs of STEC and there is a need to develop intervention strategies to reduce the entry of STEC into the food chain. The initiation of the majority of bacterial, enteric infections involves colonisation of the gut mucosal surface by the pathogen. However, probiotic bacteria can serve to decrease the severity of infection via a number of mechanisms including competition for receptors and nutrients, and/or the synthesis of organic acids and bacteriocins that create an environment unfavourable for pathogen development. The aim of the current study was to determine whether the administration of a probiotic mixture to sheep experimentally infected with a non-O157 STEC strain, carrying stx1, stx2 and eae genes, was able to decrease faecal shedding of the pathogen. The probiotic mixture contained Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus bulgaricus, Lactobacillus lactis, Streptococcus thermophilus and Enterococcus faecium. The numbers of non-O157 STEC in faecal samples collected from sheep receiving daily doses of the probiotic mixture were significantly lower at the 3rd, 5th and 6th week post-inoculation when compared to the levels recorded in untreated animals. It was concluded that administration of the probiotic mixture reduced faecal shedding of non-O157 STEC in sheep, and holds potential as a pre-harvest intervention method to reduce transmission to humans.

Antibiotic susceptibility of different lactic acid bacteria strains.

Five lactic acid bacteria (LAB) strains belonging to species Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis and Streptococcus thermophilus were tested for their susceptibility to 27 antibiotics. The minimum inhibitory concentrations of each antimicrobial were determined using a microdilution test. Among the strains a high susceptibility was detected for most of the cell-wall synthesis inhibitors (penicillins, cefoxitin and vancomycin) and resistance toward inhibitors of DNA synthesis (trimethoprim/sulfonamides and fluoroquinolones). Generally, the Lactobacillus strains were inhibited by antibiotics such as chloramphenicol, erythromycin and tetracycline at breakpoint levels lower or equal to the levels defined by the European Food Safety Authority. Despite the very similar profile of S. thermophilus LC201 to lactobacilli, the detection of resistance toward erythromycin necessitates the performance of additional tests in order to prove the absence of transferable resistance genes.

Protective effect of milk constituents and sublethal injuries limiting process effectiveness during PEF inactivation of Lb. rhamnosus.

The inactivation of Lb. rhamnosus by pulsed electric field treatment (PEF) was studied in different fractions of raw milk and Ringer solution in order to evaluate the protective effect of nutrient rich media in comparison to aqueous buffer solutions. Apart from monitoring of culturability, analysis of the physiological fitness of Lb. rhamnosus was conducted aiming to identify sublethally damaged cells. Therefore, flow cytometry and a selective medium plating technique were used and compared to each other. The goal of the study was to apply three different parameters describing the physiological fitness of the model organism Lb. rhamnosus after PEF treatment such as culturability, membrane permeability and metabolic activity depending on treatment media and parameters. A concentration dependent protective effect of the milk protein fraction could be shown and allocated to micellar casein as the major milk protein. Increasing the concentration of whey proteins up to 2% showed a similar impact on limiting the PEF inactivation of Lb. rhamnosus. The evaluation of physiological fitness of cells was based on a determination of structural and functional characteristics by rapid cellular staining using carboxyfluorescein diacetate and propidium iodide. This approach showed good accordance to the conventional selective medium plating technique for the enumeration of sublethally-injured bacteria but flow cytometry provided additional information for the characterisation of this fraction. The extent of occurrence of dead, sublethal and vital fractions of cells was found dependent on the PEF treatment parameters such as electrical field strength and energy input as well as the different milk fractions used as treatment media.