There is good evidence that the complex microbial flora present in the gastrointestinal tract of all warm-blooded animals is effective in providing resistance to disease. However, the composition of this protective flora can be altered by dietary and environmental influences, making the host animal susceptible to disease and/or reducing its efficiency of food utilization. What we are doing with the probiotic treatments is re-establishing the natural condition which exists in the wild animal but which has been disrupted by modern trends in conditions used for rearing young animals, including human babies, and in modern approaches to nutrition and disease therapy. These are all areas where the gut flora can be altered for the worse and where, by the administration of probiotics, the natural balance of the gut microflora can be restored and the animal returned to its normal nutrition, growth and health status.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2672.1989.tb05105.x

Probiotics in man and animals

Incorporation of 1.9% beta-disodium glycerophosphate (GP) into a complex medium resulted in improved growth by lactic streptococci at 30 C. The medium, called M17, contained: Phytone peptone, 5.0 g; polypeptone, 5.0 g; yeast extract, 2.5 g; beef extract, 5.0 g; lactose, 5.0 g; ascorbic acid, 0.5 g; GP, 19.0 g; 1.0 M MgSO(4).7H(2)O, 1.0 ml; and glass-distilled water, 1,000 ml. Based on absorbance readings and total counts, all strains of Streptococcus cremoris, S. diacetilactis, and S. lactis grew better in M17 medium than in a similar medium lacking GP or in lactic broth. Enhanced growth was probably due to the increased buffering capacity of the medium, since pH values below 5.70 were not reached after 24 h of growth at 30 C by S. lactis or S. cremoris strains. The medium also proved useful for isolation of bacterial mutants lacking the ability to ferment lactose; such mutants formed minute colonies on M17 agar plates, whereas wild-type cells formed colonies 3 to 4 mm in diameter. Incorporation of sterile GP into skim milk at 1.9% final concentration resulted in enhanced acid-producing activity by lactic streptococci when cells were inoculated from GP milk into skim milk not containing GP. M17 medium also proved superior to other media in demonstrating and distinguishing between lactic streptococcal bacteriophages. Plaques larger than 6 mm in diameter developed with some phage-host combinations, and turbid plaques, indicative of lysogeny, were also easily demonstrated for some systems.

Improved Medium for Lactic Streptococci and Their Bacteriophages

Phosphoenolpyruvate:carbohydrate phosphotransferase systems of bacteria.

The microbial profile of cheese is a primary determinant of cheese quality. Microorganisms can contribute to aroma and taste defects, form biogenic amines, cause gas and secondary fermentation defects, and can contribute to cheese pinking and mineral deposition issues. These defects may be as a result of seasonality and the variability in the composition of the milk supplied, variations in cheese processing parameters, as well as the nature and number of the non-starter microorganisms which come from the milk or other environmental sources. Such defects can be responsible for production and product recall costs and thus represent a significant economic burden for the dairy industry worldwide. Traditional non-molecular approaches are often considered biased and have inherently slow turnaround times. Molecular techniques can provide early and rapid detection of defects that result from the presence of specific spoilage microbes and, ultimately, assist in enhancing cheese quality and reducing costs. Here we review the DNA-based methods that are available to detect/quantify spoilage bacteria, and relevant metabolic pathways in cheeses and, in the process, highlight how these strategies can be employed to improve cheese quality and reduce the associated economic burden on cheese processors.

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Nucleic acid-based approaches to investigate microbial-related cheese quality defects

Effect of Feeding Lactobacilli on the Coliform and Lactobacillus Flora of Intestinal Tissue and Feces from Piglets 1.

The apparent instability of beta-galactosidase in toluene-treated cells or cell-free extracts of lactic streptococci is explained by the fact that these organisms do not contain the expected enzyme. Instead, various strains of Streptococcus lactis, S. cremoris, and S. diacetilactis were shown to hydrolyze o-nitrophenyl-beta-d-galactoside-6-phosphate (ONPG-6-P), indicating the presence of a different enzyme. In addition, lactose metabolism in S. lactis C(2)F was found to involve enzyme I (EI), enzyme II (EII), factor III (FIII), and a heat-stable protein (HPr) of a phosphoenolpyruvate (PEP)-dependent phosphotransferase system analogous to that of Staphylococcus aureus. Mutants of S. lactis C(2)F, defective in lactose metabolism, possessed the phenotype lac(-) gal(-). These strains were unable to accumulate (14)C-thiomethyl-beta-d-galactoside, to hydrolyze ONPG, or to utilize lactose when grown in lactose or galactose broth. In addition, these mutants contained EI and HPr, but lacked EII, FIII, and the ability to hydrolyze ONPG-6-P. This suggested that the defect was in the phosphorylation step. Lactose-negative mutants of S. lactis 7962, a strain containing beta-galactosidase, could be separated into several classes, which indicated that this organism is not dependent upon the PEP-phosphotransferase system for lactose metabolism.

Mechanisms of Lactose Utilization by Lactic Acid Streptococci: Enzymatic and Genetic Analyses

Recent literature concerning enteropathogenesis and drug resistance transfer factors in Escherichia coli are summarized as well as related papers concerning the use of antibiotics in animal feed. E. coli infection in swine (colibacillosis) also is considered, especially citations indicating the similarity between the disease in man and animals. The role of intestinal bacteria in human health is reviewed, emphasizing the importance of (a) a maintained balance of organisms in the adult, (b) breast feeding in infants to establish a large population of bifidobacteria and (c) the presence of Lactobacillus organisms to maintain healthful conditions in the human vagina. The use of Lactobacillus organisms in intestinal and vaginal disease therapy is reviewed as well as the important ecological role that lactic acid bacteria play in the natural scheme where man is concerned.

Lactic acid bacteria in food and health: a review with special reference to enteropathogenic Escherichia coli as well as certain enteric diseases and their treatment with antibiotics and lactobacilli

Citti, J. E. (Oregon State University, Corvallis), W. E. Sandine, and P. R. Elliker. β-Galactosidase of Streptococcus lactis. J. Bacteriol. 89:937–942. 1965.—Synthesis of β-galactosidase by several strains of Streptococcus lactis was induced by lactose. The rate of hydrolysis of o-nitrophenyl-β-d-galactopyranoside was used to measure enzyme activity. The enzyme of all but one strain was unstable when whole cells were sonic-treated or treated with toluene; the enzyme of one strain of S. lactis was stable to these treatments, which resulted in at least a fivefold increase in activity over that found in whole cells. The optimal assay conditions for toluene-treated cells of this strain involved incubation at 37 C in pH 7.0 sodium phosphate buffer. Lactose was the most effective inducer of enzyme synthesis. Methyl-β-d-thiogalactopyranoside, isopropyl-β-d-thiogalactopyranoside, and galactose were also inducers of the enzyme, but were not as effective as lactose. Melibiose, maltose, and calcium lactobionate were poor inducers of enzyme synthesis. Exogenously supplied glucose repressed enzyme synthesis. The means of control of induced β-galactosidase synthesis in S. lactis was similar to that in Escherichia coli.

β-Galactosidase of Streptococcus lactis

The effect of sodium fluoride on lactose metabolism and o-nitrophenyl-beta-d-galactopyranoside (ONPG) hydrolysis by Streptococcus lactis strains 7962 and C(2)F suggested that different mechanisms of lactose utilization existed in the two strains. Sodium fluoride prevented lactose utilization and ONPG hydrolysis by whole cells of S. lactis C(2)F but had no effect on S. lactis 7962. Although hydrolysis of ONPG by toluene-treated cells of S. lactis 7962 occurred without addition of phospho-enolpyruvate (PEP), toluene-treated cells of S. lactis C(2)F required the presence of this cofactor. Concentrated cell extracts of S. lactis C(2)F hydrolyzed ONPG; this hydrolysis was inhibited by NaF, but the addition of PEP, in the presence of NaF, restored maximal activity. Addition of acetyl-phosphate, carbamyl-phosphate, adenosine-5'-triphosphate, guanosine-5'-triphosphate, or uridine-5'-triphosphate did not stimulate activity. The presence of cofactors did not stimulate and NaF did not inhibit the hydrolysis in extracts of S. lactis 7962. To confirm the operation of two mechanisms, S. lactis 7962 was shown to hydrolyze lactose to glucose and galactose, whereas S. lactis C(2)F was unable to split the disaccharide. In addition, whole cells of S. lactis C(2)F rapidly accumulated a phosphorylated derivative of thiomethyl-beta-d-galactoside (TMG) which behaved chromatographically and electrophoretically like TMG-PO(4). Unexpectedly, S. lactis 7962 also accumulated a TMG derivative, although the rate was extremely low. These data indicate that different mechanisms of lactose utilization exist in the two strains, with a phosphorylation step dependent on PEP involved in S. lactis C(2)F.

P. R. Elliker

Papers

Effect of Feeding Lactobacilli on the Coliform and Lactobacillus Flora of Intestinal Tissue and Feces from Piglets 1.

Mechanisms of Lactose Utilization by Lactic Acid Streptococci: Enzymatic and Genetic Analyses

Lactic acid bacteria in food and health: a review with special reference to enteropathogenic Escherichia coli as well as certain enteric diseases and their treatment with antibiotics and lactobacilli

β-Galactosidase of Streptococcus lactis

Involvement of phosphoenolpyruvate in lactose utilization by group N streptococci.