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.
The Effect of Protein on the Thermal Death Rates of Bacteria
LANGE(1), BEHRENS(2) and HUCKEL(3) heated several kinds of bacteria to death and added them to suspensions of the living bacteria. Then they heated the mixtures and found that the admixture of the dead bacteria prolongs the thermal death time of the bacteria tested. A hypothesis was put forth by them to the effect that the added dead bacteria perhaps prevents the living ones from dying when heated. It was shown by HUCKEL that this protective activity does not depend on the kinds of the bacteria used. It would not be without interest, therefore, to ascertain whether or not the protection in question is due to protein of any kind of organism. One mg. of the bacteria cultivated for 48 hours at 28°C on agar stroke were put into l0c.c of the sterilized buffer solutions (pH=6.9) which contain 50mg., 100mg. or 200mg. of gelatin or the cells of 2 strokes killed by heating, and were shaken carefully, then put into ampullae (capacity, ca. 0.2c.c.; dia., ca. 3mm.), and were sealed. After heating at various temperatures they were cooled in water. The ampullae were disinfected with alcohol and the ends of them were cut off. Their contents were pipetted in the agar media with sterilized capillary, and the agar plate culture by the shaking method was made. The results obtained are shown in the table 1. From the table it is clear that the addition of 1-2% of gelatin prolongs about 2 times the thermal death time of the bacteria tested.
Soil organic matter and microbiological development of technosols in the South Wales Coalfield
Abstract Degraded land (formerly ‘reclaimed’) after surface-coal-mining is a serious problem in South Wales. This project uses forestation to foster soil development on opencast coal-mine spoil technosols. It compares records of humification and the soil microbiological system (microcoenosis) using paired data from the same plantings in 1998 and 2018. Statistical analysis finds no significant difference between samples collected under trees planted by the pit-planting versus trench planting method or trees planted using organic or mineral fertilizer. However, there are strong significant differences in both soil organic matter and microflora components and significant correlations with time since tree planting. Most components of the soil organic carbon (TOC) and the microflora increase, often significantly. However, compared to 2018, 1998 samples contain significantly greater maximum levels of Insoluble Carbon while the microflora had significantly more Ammonifying bacteria. Decreases in Insoluble Carbon explain the relatively small increase in TOC between 1998 and 2018. The % extracted with 0.1 N H2SO4 is significantly larger in 2018, suggesting active weathering. Similarly, both the maxima and minima of total Humic and Fulvic acids content, extracted with 0.1 M Na4P2O7 + 0.1 M NaOH, are significantly greater in 2018. Total microflora shows significant positive correlations with time-since-tree-planting as do the proportions of bacilli and actinomycetes but the correlation with ammonifying bacteria is negative. In 1998, the microflora was dominated by non-spore-forming bacteria while, in 2018, bacilli and actinomycetes prevail; both are involved in the transformation of complex organic compounds and humification. This indicates active transformation of more complex organic compounds and advancing humification. In sum, forestation of these degraded, formerly reclaimed, opencast coal-mine spoils has resulted in a massive increase in both the size and functioning of the soil organic system.