EFFECTS OF CARBOHYDRATES, PROTEINS, AND BACTERIAL CELLS IN THE HEATING MEDIA ON THE HEAT RESISTANCE OF CLOSTRIDIUM SPOROGENES
It is well known that the heat resistance of bacterial spores is influenced markedly by the nature of the medium in which the spores are heated. This fact is an important problem relating to the practical sterilization in bacteriology and in the food industry. Hence voluminous data on the heat resistance of Clostridium botulinum and Clostridium porogenes in various kinds of foods or food juices have been published (Townsend et al., 1938; Reed et al., 1951; Reynolds et al., 1952). From these data, it can be easily pointed out that the resistance value of a strain in a food is larger than that in phosphate buffer, whereas in another food the resistance value of the same strain is smaller than that in neutral phosphate buffer. Although the pH of the heating media has a marked influence on the thermal death rate of bacteria, it appears that the cause of such a difference in resistance values of a strain cannot be attributed wholly to the variation of pH values of the foods (Sognefest et al., 1948). In fact, there are evidences that some of the ingredients of food might exert a protective effect or a death accelerating effect for the organism during the heating. Weiss (1921) reported that the longer survival time was obtained when the spores of C. botulinum were suspended and heated in the foods with the higher sugar concentration. The similar protective effect of sugars on the bacterial spores has been demonstrated by Anderson et al. (1949) and Sugiyama (1951). Murray and Headlee (1931) found that the presence of peptone in the heating medium had shown sometimes a protective effect for the spores of C. tetani. Recently, Kawai and Tanaka (1952) have shown that several kinds of higher molecular substances, such as gelatin, starch, gum arabic, and alginic acid, have the protective effects for the spores of Bacillus mesentericus and Bacillus brevis. The test organisms used by these workers are relatively less resistant than the most resistant food spoilage bacteria, and the kinds of the substances tested on each strain are rather limited. Thus it seemed important to study the effects of various substances on the most resistant strain. Using the spores of Cameron's putrefactive anaerobe 3679 (Clostridium sporogenes, ATCC 7955), the effects of various carbohydrates, proteins, and bacterial cells were studied. In the previous papers Amaha (1952a,b, 1953) has clearly indicated that the thermal death time of spores of certain strains is influenced markedly by the nature of the recovery media. In general the more favorable was the nature of a medium for the development of heated spores, the longer was the survival time. Hence, it should be considered that the substances added in the basal heating medium have the two stages in their action to the spores; the first, during the heating time, and the second, after the substances were transferred in the subculture medium. In this paper an attempt was made to distinguish the two effects, i.e., the first effect, a true protection, and the second effect, a false protection, of each of substances tested.
Thermal inactivation of Bacillus cereus spores affected by the solutes used to control water activity of the heating medium.
The heat resistance of B. cereus spores (ATCC 7004, 4342 and 9818) over a wide temperature range (92-125 degrees C) in aqueous solutions of NaCl, LiCl, sucrose and glycerol at different water activities (1.00-0.71) was investigated. Sodium chloride in the heating medium tended to protect the spores of B. cereus against heat. The z-values increased significantly (P < 0.05) at and above a concentration of 4.0 M. The effects of LiCl were lower than those caused by the NaCl at the same a(w) values. An increase in z-values was observed. but the differences were only statistically significant (P<0.05) at the highest concentration tested (5.0 M). A concentration of sucrose 0.87 M caused in all cases a reduction in D-values, which was most pronounced for strains 4342 and 9818. With increasing concentration of sucrose ( > 0.87 M), the D-values showed an increase, although only those obtained for strain 4342 in sucrose solutions 2.22 M were higher than those found in pure water. The z-values were significantly higher (P < 0.05) when sucrose was added at concentrations above 1.42 M, except for strain 4342. When a(w) was lowered from 0.96 to 0.71 with glycerol, D-values obtained gradually increased, about 30, 50 and 60 fold for 4342, 7004 and 9818 strains, respectively. No significant effect on z-values were detected.
Inhibition of Root Growth and Polyamine Metabolism in Sunflower (Helianthus annuus) Seedlings Under Cadmium and Copper Stress
Although sunflower is usually regarded as a highly tolerant crop, impairment of root growth at initial stages of plant development may result in poor crop establishment and higher susceptibility to pathogen attack. In order to evaluate if Cd2+ and Cu2+ may impact on sunflower germination and initial root development, a pot experiment under controlled conditions was carried out. Possible involvement of polyamine metabolism in sunflower response to these stressors was also investigated. Although Cd2+ and Cu2+ treatments affect neither seed germination nor radicle emergence, sunflower seedlings grown in the presence of these heavy metals showed significant inhibition of root growth, being this inhibition greater for Cd2+. Both metals caused significant increases in proline contents at the highest concentrations tested (0.5 and 1 mM), and these increments were more pronounced for Cd2+ treatments, especially between days 3 and 10. Metals also increased putrescine (Put) contents at all concentrations assayed from the seventh day onward, causing no variations on this polyamine time-course pattern. Spermine and spermidine contents, however, were increased only by 1 mM Cd2+. Arginine decarboxylase seems to have been the enzyme responsible for Put increases under both metal treatments. This work demonstrates that initial root growth of sunflower seedlings may be significantly impaired in Cd2+ or Cu2+ contaminated soils. It also shows that polyamines are key biological compounds, which are probably involved in signaling pathways triggered under stress environmental conditions.