• Enterococci in Food Fermentations: Functional and Safety Aspects

      Cogan, Tim; Rea, Mary; Drinan, Finbarr; Gelsomino, R. (Teagasc, 2001-04-01)
      Enterococci are natural residents of the human and animal gastrointestinal tracts; many species are also found in soil, plants and food. These organisms also form an important part of the microflora of many cheeses, especially those made in Southern Europe, where they can reach levels of 107 - 108 cfu/g. There is contradictory information on their role in flavour development in cheese with some studies showing that they have a positive effect and others a negative one. Enterococcus faecalis, Ec. faecium and Ec. durans are the important species found in cheese, though recent results from our laboratory show that Ec. casseliflavus may also be important (see below). Many of these species withstand pasteurisation. Their presence in food has been questioned because they are responsible for many nosocomial infections in hospitals. They are also promiscuous and easily transfer antibiotic resistance to other organisms and acquire resistance to vancomycin themselves. Cheddar cheese has a complex microflora and is conducive to growth of many bacteria, especially lactic acid bacteria. Enterococci are facultative anaerobes, which ferment lactose and can grow in high salt concentrations. Therefore, they should grow in cheese if they are present in the raw milk. Phenotypically they can be confused with starter lactococci. Traditionally, they are separated from lactococci by their ability to grow at 45°C and in 6.5% salt. However, these tests have serious drawbacks since some species of enterococci cannot grow at 45°C and some lactococci can grow at 45°C and in 6.5% salt. The effect of enterococci on flavour development in Cheddar cheese has not been studied to any great extent. The overall objectives of this collaborative project were to investigate the taxonomic relationships between food, veterinary and clinical isolates of enterococci, their virulence, their ability to produce toxins, their antibiotic resistance and their technological performance in cheesemaking. The specific objectives of the Moorepark team were to study the co-metabolism of citrate and sugar by enterococci, develop a DNA probe to distinguish between Enterococcus and Lactococcus and evaluate the contribution of enterococci to flavour development in Cheddar cheese.
    • Role of Lactobacilli in Flavour Development of Cheddar Cheese.

      Beresford, Tom; Cogan, Tim; Rea, Mary; Drinan, Finbarr; Fitzsimons, Nora; Brennan, N.; Kenny, Owen; Fox, P.F. (Teagasc, 2001-05-01)
      Cheddar cheese is a complex microbial ecosystem. The internal cheese environment, in particular of hard and semi-hard cheeses, is not conducive to the growth of many microorganisms. At the beginning of ripening the dominant microorganisms are the starter bacteria which are present at high levels (~109/g). However, during ripening, non-starter lactic acid bacteria (NSLAB) grow from relatively low levels (<103/g) at the beginning of ripening, to 108/g within 6 - 8 weeks. Other bacteria, e.g. enterococci and staphylococci, may also be present but in much lower numbers. In a previous study of mature and extra mature Cheddar cheeses from different manufacturers (see End of Project Report No. 1), it was found that the NSLAB population was dominated by strains of Lb. paracasei. However, their contribution to cheese flavour and their source(s) are still unclear, nor is it known if the NSLAB flora is unique to each plant. Hence, understanding the growth of this group of organisms in cheese is a key to defining their role in flavour development. The biochemistry of flavour development in cheese is poorly understood. For most cheese varieties, including Cheddar, proteolysis, which results in the accumulation of free amino acids, is of vital importance for flavour development. Increasing evidence suggests that the main contribution of amino acids is as substrates for the development of more complex flavour and aroma compounds. The manner by which such compounds are generated in cheese is currently the focus of much research. Starter bacteria have been shown to contain a range of enzymes capable of facilitating the conversion of amino acids to potential flavour compounds. However, the potential of lactobacilli (NSLAB) to produce similar enzymes has only recently been investigated. Hence, although, it is generally accepted that the cheese starter flora is the primary defining influence on flavour development, the contribution of NSLAB is also considered significant. The objectives of these studies were: - to develop a greater understanding of the behaviour of NSLAB in cheese, and - to identify suitable strains, and other cheese bacteria, to be used as starter adjuncts for flavour improvement.
    • Use of Bacteriocins to Improve Cheese Quality and Safety

      Ross, R Paul; Hill, Colin; Ryan, Maire; Cunniffe, Alan; McAuliffe, Olivia; Murray, Deirdre; O'Keefe, Triona; Rea, Mary (Teagasc, 1998-09-01)
      The objectives of this project were to generate, characterise and exploit a range of novel bacteriocin producing starter cultures to improve both the safety and the quality of fermented dairy foods. The main conclusions were as follows: Lacticin 3147 is a broad spectrum bacteriocin which inhibits a wide range of Gram-positive bacteria including lactobacilli, clostridia and Listeria. The bacteriocin has been purified by chromatographic procedures and has been shown to be composed of two peptides, both of which are required for biological activity. The mechanism of action of lacticin 3147 has been elucidated. The entire plasmid encoding lacticin 3147 has been sequenced and the bacteriocin in distinct from any previously characterised lactococcal bacteriocin. The Food Grade introduction of the bacteriocin genes into cheese starters was carried out. Lacticin 3147 producing starters have been used to control the pathogen Listeria monocytogenes on the surface of mould ripened cheese. Lacticin 3147 producing starters have been used to control the non-starter lactic acid bacteria complement in Cheddar cheese during the ripening process. A novel starter system using a bacteriocin (lactococcin)- producing adjunct has been designed which gives increased cell lysis during Cheddar cheese manufacture while ensuring that efficient acid production is not compromised. In summary these studies have found that naturally occurring antimicrobials such as bacteriocins have a wide range of applications in the food industry for improving both the quality and safety of fermented dairy products.