The synergistic effect of nisin and lactoferrin on the inhibition of Listeria monocytogenes and Escherichia coli O157:H7.

AIMS: The goal of this study was to determine whether nisin and lactoferrin would act synergistically to inhibit the growth of Listeria monocytogenes and Escherichia coli O157:H7. METHODS AND RESULTS: Lactoferrin and nisin separately or in combination were suspended in peptone yeast glucose broth and following inoculation with L. monocytogenes or E. coli O157:H7 growth inhibition of each pathogen was determined. At 1000 microg ml(-1) lactoferrin L. monocytogenes was effectively inhibited. However, E. coli O157:H7 initially was inhibited and then grew to cell density similar to the control. A combination of 500 microg ml(-1) of lactoferrin and 250 IU ml(-1) of nisin effectively inhibited the growth of E. coli O157:H7, whereas, 250 microg ml(-1) of lactoferrin and 10 IU ml(-1) of nisin were inhibitory to L. monocytogenes. CONCLUSIONS: The results suggest that lactoferrin and nisin act synergistically to inhibit the growth of L. monocytogenes and E. coli O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: Natural preservatives that are active against gram-positive and gram-negative pathogens are desirable to the food industry and consumers. This study demonstrates that lactoferrin and nisin work synergistically reducing the levels required independently inhibiting growth of two major foodborne pathogens. Previous reported results indicated a low level of antimicrobial activity; however, this work was not performed in low divalent cation concentration media. It has been suggested that nondivalent cation-limiting medium such as trypticase soy broth (TSB), can reduce or completely eliminate the inhibitory activity. Further knowledge of these interactions can increase the understanding of the antimicrobial activity of lactoferrin. This should make the use of these compounds by industry more attractive.

Antibacterial activity of pepsin-digested lactoferrin on foodborne pathogens in buffered broth systems and ultra-high temperature milk with EDTA.

AIMS: To evaluate the antimicrobial activity in peptone yeast extract glucose (PYG) broth and ultra-high temperature (UHT) milk of bovine lactoferrin hydrolysate (LFH) with pepsin against the foodborne pathogens Salmonella Stanley, Escherichia coli, Listeria monocytogenes and Staphylococcus aureus. METHODS AND RESULTS: The LFH was suspended in PYG and the minimum inhibitory concentration for each pathogen determined. The LFH was also suspended in UHT milk adjusted to pH 4 or 7, samples incubated at 4 or 35 degrees C and the change in bacterial cell population determined. Experiments in UHT milk were conducted using L. monocytogenes and E. coli O157:H7. At pH 4 LFH reduced the population of E. coli O157:H7 and L. monocytogenes by approx. 2 log; however, only E. coli O157:H7 was inhibited in samples adjusted to pH 7. The addition of EDTA (10 mg ml(-1)) to UHT milk supplemented with LFH did not markedly influence the growth of E. coli O157:H7 or L. monocytogenes. CONCLUSIONS: The results suggest that, under low pH and refrigeration conditions, LFH can limit the growth or reduce the population of pathogenic bacteria in a dairy product. SIGNIFICANCE AND IMPACT OF THE STUDY: Natural preservatives that are active against Gram-negative and Gram-positive bacteria are desirable to the food industry. This study demonstrates that LFH is effective in a complex food system. Moreover, the LFH used was not purified, making its use by industry more attractive.

Effects of the nitro-group on the mutagenicity and toxicity of some benzamines.

The Ames Salmonella/microsomal assay was employed to test the mutagenicity of some benzamines (aniline, and o- and p-phenylenediamine) and their nitro-derivatives (p-nitroaniline, 2-nitro-p-phenylenediamine, 3- and 4-nitro-o-phenylenediamine), using strains TA98 and TA100 and their nitroreductase-deficient mutants, TA98NR and TA100NR, in the presence and absence of rat S9 mix. The addition of the nitro-group to benzamine molecules converted them into direct mutagens. Furthermore, the position of the nitro-group affected their mutagenic activities. Cytotoxicity testing with Chinese hamster ovary cells (CHO-K1) showed that the presence of the nitro-group in these compounds had no specific effect on toxicity. The test compounds all showed a dose-related increase in inducing chromosomal aberrations in CHO cells. However, the presence of the nitro-group did not affect potency in inducing chromosomal aberrations. Compounds containing the nitro-group had higher initial oxidation potentials and dipole moments (mu) than their nonnitro-containing counterparts. The mutagenicity and toxicity of these compounds were not related to physico-chemical properties, including oxidation potential, energy difference (deltaE) between the lowest unoccupied molecular orbital (LUMO) and the highest occupied molecular orbital (HOMO), ionization potential (I.P.), and mu.

Mutagenicity and toxicity studies of p-phenylenediamine and its derivatives.

The mutagenicity of p-phenylenediamine and its derivatives was tested using Ames Salmonella strains TA98 and TA100. p-Phenylenediamine was weakly mutagenic to TA98 with metabolic activation. 2-Nitro-p-phenylenediamine was directly mutagenic to both strains, while 2-methyl-p-phenylenediamine required S9 mix. All the test compounds induced a dose-related increase in chromosomal aberrations in Chinese hamster ovary (CHO) cells in the absence of the S9 mix. The mutagenicity and toxicity of these compounds did not correlate with their oxidation potentials, or any other tested physicochemical properties including the energy difference between the lowest unoccupied and the highest occupied molecular orbital, ionization potential, and dipole moment.

Simultaneous uptake of galactose and glucose by Azotobacter vinelandii.

Azotobacter vinelandii growing on galactosides induced two distinct permeases for glucose and galactose. The apparent Vmax and Km of the galactose permease were 16 nmol galactose/min per 10(10) cells and 0.5 mM, respectively. The apparent Vmax and Km of the glucose permease were 7.8 nmol glucose/min per 10(10) cells and 0.04 mM, respectively. Excess glucose had no effect on the galactose uptake. However, excess galactose inhibited glucose transport. The galactosides-induced glucose permease also exhibited different uptake kinetics from that induced by glucose.

Exogenous fibronectin requirement for adhesion by neoplastic human cells.

We have studied the effect of serum, plasma and plasma fibronectin on the spreading behavior of several normal and neoplastic human cells using a quantitative spreading assay. Normal human cells are rich in surface fibronectin and are able to spread in the absence of serum or exogenous fibronectin. Virus-transformed cells and cells of neoplastic origin have reduced or no recognizable surface fibronectin and are dependent on serum or plasma for spreading. In agreement with previous studies on rodent cells, fibronectin appears to be the major factor responsible for this effect. Neoplastic cells can spread on fibronectin-coated substrata or can recruit fibronectin directly from the medium. The capacity of transformed cells to recruit fibronectin from the medium correlates with the presence of intercellular fibronectin as seen by the immunofluorescence technique on monolayer cultures. Our results are in keeping with the concept that neoplastic cells may be dependent on extracellular fibronectin for adherence in vivo and that this dependency may vary between different cell types.