Satoshi Morozumi

Modeling Surface Growth of Escherichia coli on Agar Plates.

ABSTRACT Surface growth of Escherichia coli cells on a membrane filter placed on a nutrient agar plate under various conditions was studied with a mathematical model. The surface growth of bacterial cells showed a sigmoidal curve with time on a semilogarithmic plot. To describe it, a new logistic model that we presented earlier (H. Fujikawa et al., Food Microbiol. 21:501-509, 2004) was modified. Growth curves at various constant temperatures (10 to 34°C) were successfully described with the modified model (model III). Model III gave better predictions of the rate constant of growth and the lag period than a modified Gompertz model and the Baranyi model. Using the parameter values of model III at the constant temperatures, surface growth at various temperatures was successfully predicted. Surface growth curves at various initial cell numbers were also sigmoidal and converged to the same maximum cell numbers at the stationary phase. Surface growth curves at various nutrient levels were also sigmoidal. The maximum cell number and the rate of growth were lower as the nutrient level decreased. The surface growth curve was the same as that in a liquid, except for the large curvature at the deceleration period. These curves were also well described with model III. The pattern of increase in the ATP content of cells grown on a surface was sigmoidal, similar to that for cell growth. We discovered several characteristics of the surface growth of bacterial cells under various growth conditions and examined the applicability of our model to describe these growth curves.

Inhibitory effects of condiments and herbal drugs on the growth and toxin production of toxigenic fungi.

The effects of thirteen kinds of powdered herbal drugs and seven kinds of commercial dry condiments on the growth and toxin production ofAspergillus parasiticus, A. flavus,A. ochraceus, andA. versicolor were observed by introducing these substances into culture media for mycotoxin production.Of the twenty samples tested, cinnamon bark completely inhibited the fungal growth, while the others only inhibited the toxin production.The inhibitors were easily extracted from the samples with solvents such as hot water, chloroform, or ethanol.The extracts from coptis, philodendron bark, mustard, green tea leaves, and zanthoxylum completely inhibited the aflatoxin production ofA. parasiticus, however, they had little or no inhibitory effect againstA. flavus.

Fungal contamination and mycotoxin-producing potential of dried beans

A total of 604 samples of about 7 different types of beans was examined to determine their mycological profiles, and suitability for use as solid substrates for mycotoxin production.All of the samples were collected from bean jam makers in Tokyo by the official food examiners.Genera Penicillium and Aspergillus were predominant, and genus Wallemia was also found commonly in all types of beans.Mycotoxin-producing Aspergillus strains were isolated from 52 samples of beans, approximately 9% of the total. The highest incidence of toxigenic Aspergillus (14.1%) was found in kidney beans. Red beans and peas inoculated with Aspergillus ochraceus were found to produce about 7 to 8 times more toxin than was obtained in a liquid medium, and red beans inoculated with A. versicolor produced more toxin than was obtained in yeast extract sucrose broth. Green peas inoculated with Fusarium graminearum produced about 8 times more T-2 toxin than was obtained in 1% peptone containing Czapek solution under comparable culture conditions.

Comparison of capillary and test tube procedures for analysis of thermal inactivation kinetics of mold spores.

Characteristics of capillary and test tube procedures for thermal inactivation kinetic analysis of microbial cells were studied for mold spores. During heating, capillaries were submerged in a water bath and test tubes were held with their caps positioned above the level of the heating medium. Thermal inactivation curves of Aspergillus niger spores in capillaries at around 60 degrees C consisted of a shoulder and a fast linear decline, whereas curves in test tubes consisted of a shoulder, a fast linear decline, and a horizontal tail. There were no significant differences in values of the rate and the delay of fast declines in curves between the procedures. Some experiments were done to clarify the cause for tailing with test tubes. There were no tails with test tubes whose inner walls were not contaminated by A. niger spores, suggesting that tails arise from A. niger spores contaminating the inner walls of test tubes. Temperature of the inner wall at the level of a heating medium was lower than that of the medium. Further, there were no tails for test tubes submerged in the heating medium. These results showed that the reason for survival of contaminants on the upper wall of test tubes was that cells were not subjected to sufficient inactivation temperature. Finally, thermal inactivation curves of A. niger spores in capillaries at various constant temperatures were studied. Curves consisted of a shoulder and a fast linear decline at 57 degrees C and above, whereas curves at below 57 degrees C consisted of a shoulder, a fast linear decline, and a sloping tail.

Chemically defined medium for high yields of sterigmatocystin.

Isolate of Aspergillus versicolor strain produced 138 μg/ml of sterigmatocystin in a complete synthetic medium containing sucrose, salts, 1-phenylalanine, and Ca-pantothenate. The SSP (sucrose salts phenylalanine) medium apparently provided all necessary ingredients for the production of high levels of sterigmatocystin. For optimal sterigmatocystin formation, the amounts of sucrose and 1-phenylalanine were found to be 200 g and 5 g per liter, respectively. When Ca-pantothenate (0.01 g per liter) added, much higher amounts of sterigmatocystin were recovered, whereas CaCl2 addition (0.01%) drastically reduced the yield. The high levels of sterigmatocystin were recovered in the cultures which incubated stationarily at 26 to 29 °C for over 12 days. Seven strains or isolates tested yielded high levels of sterigmatocystin in the SSP medium, whereas in each other media such as YES medium and rice medium only one isolate yielded highest amount of sterigmatocystin was found.

Growth of moulds inoculated into commercial mineral water

The growth of mould spores of Penicillium sp. and Cladosporium sp. inoculated in a commercial mineral water product was studied. The strains had been isolated as fungal foreign bodies in commercial mineral waters. In product A, which was not originally sterilized and was contaminated with psychrophilic bacteria, the inoculated mould spores of the strains did not grow; no increases in viable colony counts or β‐glucans concentration in the samples were observed during storage. In a sterilized product A, inoculated spores of the strains grew into visible foreign bodies. The viable colony counts and the β‐glucans concentration in the samples increased during storage. These results showed that in a sterilized mineral water product, mould spores could grow into visible foreign bodies.

Microbial Flora and Organic Acid Contents in “Tea Fungus”

To evaluate the safety of biological and chemical contents in “Tea fungus” as a kind of soft drink, microbial flora and organic acid content in this drink were investigated.As the result of microbial investigation, Acetobacter xylinum was isolated from the drink as sole source of bacteria and other yeasts such as Saccharomyces cerevisiae, S. inconspicus, Candida tropicalis, Debaryomyces hansenii, were also found. Among those isolates tested, A. xylinum was found to be a zoogloeal mats forming organism, and the product was increased in cooperation with S. cerevisiae.As the result of organic acid analysis by gas liquid chromatography, acetic acid, formic acid and lactic acid were found, especially acetic acid content occupied more than 95% of the total of them in this drink.Several kinds of pathogenic bacteria such as Salmonella typhi, Shigella sonnei, pathogenic Escherichia coli, Staphylococcus aureus, were all died within 24 hours, when experimentally inoculated into the drink and incubation at both of 25°C and 37°C, though several kinds of filamentous fungi were found grow well in it.