Influence of Carbohydrate and Nitrogen Source on Patulin Production by Penicillium patulum

A strain of Penicillium patulum, isolated from cheddar cheese, produced patulin when grown on liquid media containing lactose and milk nitrogen sources. Patulin production was affected by the temperature of incubation, the type and amount of carbohydrate, and the type of nitrogen source present. Patulin levels generally were depressed by incubation at 5 C and low carbohydrate levels. Patulin was produced at low levels in the absence of sugars at 5 C when the mold was grown on milk nitrogen sources. No patulin was detected in cultures grown on 25% casein slurries or cheddar cheese, even though growth of the mold was extensive.     

The importance of combining air sampling and surface analysis when studying problematic houses for mold biodiversity determination

Houses that underwent water damages are often responsible for heath problems of the occupants. Since there is no universally used protocol for the analysis, we wanted to verify the usefulness of surface sampling versus air sampling for the evaluation of mold diversity in problematic houses and the value of the number of visible mold growth zones to predict air quality. Seventeen houses were sampled for culturable molds in the air and on the surfaces showing contamination. We compared the mold taxa found in the air and on the surfaces and verified the correlation between the number of moldy surfaces and airbone mold concentration. This study demonstrated that, surprisingly, some of the so called wet spore molds (e.g. Stachybotrys) were found more often from air than surface samples whereas, some dry spore molds (e.g. Asp. fumigatus) was more easily isolated from surface samples. There was a good correlation between the number of visible mold growth zones and the concentration of airborne molds. We conclude that air and surface sampling are necessary to evaluate mold diversity in problematic houses and that the number of mold growth zones is a good predictor of airborne mold concentration.     

Effect of Environmental and Physiological Conditions on the Phase of Adjustment of Pseudomonas fragi

The duration of the phase of adjustment of Pseudomonas fragi was affected by the physiological age and growth temperature of the inoculum, as well as by the temperature at which the growth curve was determined. Cultures in the exponential phase of growth gave shorter lags than stationary-phase and resting-phase inocula. Inocula from the latter phase gave the longest lags. Inocula grown at the temperature at which the growth curve was determined usually gave the shortest lags: the greater the difference between the incubation temperature of the inoculum and the incubation temperature of the growth curve, the longer the lag. Inocula grown at temperatures below the incubation temperature of the culture tended to produce longer lags than inocula grown at temperatures above the incubation temperature. The combined effect of physiological age and incubation temperature of the inoculum was additive. The effect of the incubation temperature of the culture upon the duration of the lag depended upon the method used to determine this phase. Lags that were measured in physical time (i.e., Lockhart's lag) decreased as the incubation temperature of the culture was increased, within the temperatures used. But Monod's lag, which measures physiological time, did not decrease as the temperature of growth increased but rather appeared to vary around some constant value dependent upon the physiological condition of the culture.     

Unusual polyamines in slime molds Physarum polycephalum and Dictyostelium discoideum

We analyzed the cellular contents of not only major polyamines but also minor polyamines in slime molds Physarum polycephalum and Dictyostelium discoideum. The presence of putrescine and spermidine in either plasmodia or myxamoebae of these molds as major polyamines was confirmed. In addition to these polyamines, appreciable amounts of 1,3-diaminopropane were detected in P. polycephalum and D. discoideum. Cadaverine and sym-homospermidine were detected in P. polycephalum even when the slime mold was cultured in a chemically defined growth medium. Spermine was not detected when these molds were grown in synthetic media. Other "unusual" polyamines such as norspermidine, norspermine, thermospermine, aminopropylcadaverine, and canavalmine were not detected in either mold.     

Combining heat treatment and subsequent incubation temperature to prevent growth from spores of non-proteolytic Clostridium botulinum

Refrigerated processed foods of extended durability rely on a mild heat treatment combined with refrigerated storage to ensure microbiological safety and quality. The principal microbiological safety risk in foods of this type is non-proteolytic Clostridium botulinum. In this article the combined effect of mild heat treatment and refrigerated storage on the time to growth and probability of growth from spores of non-proteolytic Cl. botulinum is described. Spores of non-proteolytic Cl. botulinum (two strains each of type B, E and F) were heated at 90°C for between 0 and 60 min and subsequently incubated at 5°, 10° or 30°C in PYGS broth in the presence or absence of lysozyme. The number of spores that resulted in turbidity depended on the combination of heat treatment, incubation time and incubation temperature they received. Heating at 90°C for 1 or more min ensured a 10⁶ reduction when spores were subsequently incubated at 5°C for up to 23 weeks. Heating at 90°C for 60 min ensured a 10⁶ reduction over 23 weeks when subsequent incubation was at 10°C in the presence of added lysozyme. The same treatment did not reduce the spore population by 10⁶ when subsequent incubation was at 30°C.     

Vigorous Mold Growth in Soils After Addition of Water-Insoluble Fatty Substances

Various water-insoluble fatty compounds, when added to soil in finely divided form, will support as high-caloric nutrients a visible, vigorous growth of the molds, Fusarium solani Mart., F. diversisporum Sherb., and F. equiseti. n-Paraffins and olefins are most effective, because the effect of additives is reduced to the extent that oxygen atoms are introduced into the molecule. n-Fatty alcohols support growth in soil almost as well as the paraffins; however, growth is reduced when branched-chain compounds are added as nutrients. Compounds that will support mold growth when added to air-dried soil as finely powdered solids will not do so when incorporated at temperatures above their melting point, but will produce dense growth when applied to wet soil in this form. Mold growth is correlated with degradation of fatty matter. The rate of degradation is controlled by the availability of water, oxygen, and the basic inorganic nutrients.     

Diversity and significance of mold species in Norwegian drinking water

In order to determine the occurrence, distribution, and significance of mold species in groundwater- and surface water-derived drinking water in Norway, molds isolated from 273 water samples were identified. Samples of raw water, treated water, and water from private homes and hospital installations were analyzed by incubation of 100-ml membrane-filtered samples on dichloran-18% glycerol agar. The total count (number of CFU per 100 ml) of fungal species and the species diversity within each sample were determined. The identification of mold species was based on morphological and molecular methods. In total, 94 mold species belonging to 30 genera were identified. The mycobiota was dominated by species of Penicillium, Trichoderma, and Aspergillus, with some of them occurring throughout the drinking water system. Several of the same species as isolated from water may have the potential to cause allergic reactions or disease in humans. Other species are common contaminants of food and beverages, and some may cause unwanted changes in the taste or smell of water. The present results indicate that the mycobiota of water should be considered when the microbiological safety and quality of drinking water are assessed. In fact, molds in drinking water should possibly be included in the Norwegian water supply and drinking water regulations.     

Effect of Processing on the Microflora of Norwegian Seaweed Meal, with Observations on Sporendonema minutum (Høye) Frank and Hess

Meal from the brown seaweed Ascophyllum nodosum (L.) Le Jol. is mainly used as an animal feed supplement. Since moist weed often develops a marked mold growth and since little was known about the microflora of seaweed meal, a cultural procedure was developed to enumerate the populations of bacteria, yeasts, and molds of seaweed meals manufactured by different drying processes. The microflora could be supported by a variety of media varying in levels of nutrition and in the source and concentration of salts. Fresh weed contained less than 10(3) bacteria and less than 10(2) yeasts and molds per g (dry weight). The type and extent of microbial populations in seaweed meal appeared to be dependent upon the method of seaweed drying. Rotary drum-drying at temperatures decreasing from 800 to 80 C maintained or reduced the microbial populations to 10(3) organisms per g (dry weight). Although meals with high nutritional quality can be obtained with warm air- or rock-dried weed, these conditions can also permit bacterial and mold development. Extended rock-drying in variable weather conditions and prolonged storage of moist weed, both of which decrease the nutritional quality, also lead to high bacterial numbers and to a marked development of the halophilic brown mold Sporendonema minutum which attained populations of 10(8) viable spores per g of dried weed. A poultry diet containing 5% badly molded weed had no apparent toxic or growth-depressing effect when fed to chicks.     

Diversity and Significance of Mold Species in Norwegian Drinking Water

In order to determine the occurrence, distribution, and significance of mold species in groundwater- and surface water-derived drinking water in Norway, molds isolated from 273 water samples were identified. Samples of raw water, treated water, and water from private homes and hospital installations were analyzed by incubation of 100-ml membrane-filtered samples on dichloran-18% glycerol agar. The total count (number of CFU per 100 ml) of fungal species and the species diversity within each sample were determined. The identification of mold species was based on morphological and molecular methods. In total, 94 mold species belonging to 30 genera were identified. The mycobiota was dominated by species of Penicillium, Trichoderma, and Aspergillus, with some of them occurring throughout the drinking water system. Several of the same species as isolated from water may have the potential to cause allergic reactions or disease in humans. Other species are common contaminants of food and beverages, and some may cause unwanted changes in the taste or smell of water. The present results indicate that the mycobiota of water should be considered when the microbiological safety and quality of drinking water are assessed. In fact, molds in drinking water should possibly be included in the Norwegian water supply and drinking water regulations.     

Effect of Processing on the Microflora of Norwegian Seaweed Meal,with Observations on Sporendonema minutum (H?ye) Frank and Hess

Meal from the brown seaweed Ascophyllum nodosum (L.) Le Jol. is mainly used as an animal feed supplement. Since moist weed often develops a marked mold growth and since little was known about the microflora of seaweed meal, a cultural procedure was developed to enumerate the populations of bacteria, yeasts, and molds of seaweed meals manufactured by different drying processes. The microflora could be supported by a variety of media varying in levels of nutrition and in the source and concentration of salts. Fresh weed contained less than 10³ bacteria and less than 10² yeasts and molds per g (dry weight). The type and extent of microbial populations in seaweed meal appeared to be dependent upon the method of seaweed drying. Rotary drum-drying at temperatures decreasing from 800 to 80 C maintained or reduced the microbial populations to 10³ organisms per g (dry weight). Although meals with high nutritional quality can be obtained with warm air- or rock-dried weed, these conditions can also permit bacterial and mold development. Extended rock-drying in variable weather conditions and prolonged storage of moist weed, both of which decrease the nutritional quality, also lead to high bacterial numbers and to a marked development of the halophilic brown mold Sporendonema minutum which attained populations of 10⁸ viable spores per g of dried weed. A poultry diet containing 5% badly molded weed had no apparent toxic or growth-depressing effect when fed to chicks.     

Cold shock during liquid production increases storage shelf-life of Cryptococcus nodaensis OH 182.9 after air-drying

Fermentation, formulation and drying studies are necessary and important in order to simplify production, transportation, storage and application of biocontrol agents. Air-drying is a convenient and economical drying method for developing microbial biocontrol products. Experiments were designed to determine the effect of temperature shock during liquid cultivation on cell survival of a Fusarium head blight biocontrol agent Cryptococcus nodaensis OH 182.9 after air-drying. OH 182.9 cultures were grown at various temperatures in semi-defined complete liquid media, with cultures grown at 25°C for 48 h serving as the standard control culture condition. Harvested cultures were mixed with 10% diatomaceous earth (DE), vacuum filtered, air dried for 20 h at 60-70% RH, and stored at 4°C. In general, cells grown at 25°C for 20 h followed by cultivation at 15°C for 28 h survived air-drying better than control cells. The survival of cells subjected to heat shock at 31°C generally did not differ from control cells regardless of whether heat shock was applied at the late exponential or early stationary stage of growth. In another experiment designed to optimize the effect of cold temperatures during cultivation on subsequent survival of air-dried cells in DE at 4°C and room temperature (25°C), prolonged (28 h) cold shock at 10 and 15°C after incubation at 25°C for 20 h enhanced the storage stability (shelf-life) of a DE-formulated OH 182.9 product. In greenhouse tests, air-dried cells of OH 182.9 stored for 6 weeks at 4°C maintained a higher biocontrol efficacy than cells stored for 6 weeks at 25°C.     

The effect of fungal species on the fluorescent lectin test.

Fungal (mold) contamination is an important indicator of low-quality raw product used in food processing operations. Fluorescent-labeled lectins, specific for chitin, have been shown to be valuable for quantitative detection of mold in raw tomatoes.In this research, the response of individual fungal species to a rapid fluorescent lectin assay was investigated. Ten of the most common mold species were grown on two types of artificial broth media, and added to blended field tomatoes. The assay was conducted on each species, and linear regressions were developed, comparing the fluorescent lectin assay score with the fungal dry weight. The assay was able to detect all molds at sensitivities required for the tomato industry, and had high linearity (r2 ranging from 0.72 to 0.99) and low variability (standard error of calibration ranging from 20 to 116 microg of fungal biomass/ml of tomato juice) for individual species grown on V-8 juice broth.     

Distribution of Neuraminidase among Food-poisoning Strains of Clostridium perfringens

A survey was made to determine the distribution of the enzyme neuraminidase among 76 strains of Clostridium perfringens. Representative strains from each toxigenic type (A to F) and atypical C. perfringens type A food-poisoning strains of both American and English (Hobbs types) origin were tested. Both the American food-poisoning and nonfood-poisoning associated cultures consisted of both neuraminidase-positive and -negative strains. Furthermore, American strains which could not be differentiated from the original Hobbs cultures consisted of both neuraminidase-positive and -negative representatives. In contrast, the English (Hobbs) strains uniformly failed to produce an active intracellular or extracellular neuraminidase. No enzyme activity was detected in these strains when cultures were grown in different growth media, when grown in the presence of substrate (neuraminlactose), or upon extended incubation of enzyme preparations with substrate. With the exception of a type F strain, representative strains of the other toxigenic types (A to F) produced neuraminidase; 85% of the typical type A strains contained the enzyme.     

Effect of growth conditions and storage on the specific activity of beta-lactamases of Bacteroides spp

The influences of growth conditions and cold storage on the specific activity of beta-lactamases of four strains of Bacteroides spp. was studied. Interbatch variation was observed in extracts prepared in an identical way on separate occasions but less variation was observed in extracts prepared from bacteria grown on Brain Heart Infusion agar supplemented with yeast extract, haemin and menadione, than in similar extracts of bacteria grown in broth or on other solid media. The loss of enzyme activity seen during the stationary phase of growth of some strains in broth was minimal during incubation for 48 h on agar. Storage of enzyme extracts at 4 degrees C was associated with loss of enzyme activity, but activity was retained during storage at -70 degrees C for up to 32 days. Freezing and thawing had little effect on enzyme activity.     

Some Cultural Conditions That Control Biosynthesis of Lipid and Aflatoxin by Aspergillus parasiticus

Synthesis of total lipid and aflatoxin by Aspergillus parasiticus as affected by various concentrations of glucose and nitrogen in a defined medium and by different incubation temperatures was studied. Maximal yields of lipid and aflatoxin were obtained with 30% glucose, whereas mold growth, expressed as dry weight, was maximal when the medium contained 10% glucose. Maximal mold growth occurred when the medium contained 3% (NH(4))(2)SO(4); however, 1% (NH(4))(2)SO(4) favored maximum accumulation of lipid and aflatoxin. Growth of mold and synthesis of lipid and toxin also varied with the incubation temperature. Maximal mold growth occurred at 35 C, whereas most toxin appeared at 25 C. Maximal production of lipid occurred at 25 and 35 C but production was more rapid at 35 C. Essentially all glucose in the medium (5% initially) was utilized in 3 days at 25 and 35 C but not in 7 days at 15 and 45 C. Patterns for formation of lipid and aflatoxin were similar at 15 and 25 C when a complete growth medium was used and at 28 C when the substrate contained various concentrations of glucose or (NH(4))(2)SO(4). They were dissimilar when the mold grew at 35 or 45 C. At these temperatures lipid was produced preferentially and only small amounts of aflatoxin appeared.     

Effect of some nutrients on the production of cyclopiazonic acid by Penicillium verrucosum var. cyclopium

Summary The effect of some sugars in different concentrations and some nitrogeneous organic constituents in 2% yeast extract (basal medium) on production of cyclopiazonic acid (CA) by Penicillium verrucosum var. cyclopium was studied at room temperature. Maximum CA production was observed after 14 days in a medium containing 2% yeast extract +2.5% sucrose. Ammonium lactate had a negative effect on the production of CA by the test culture. Nitrogeneous organic constituents such as peptone and tryptone did not enhance the yield of CA in the medium. After an initial drop in the pH, a general increase in pH was observed as the incubation time increased. Curdled milk was also found to be a suitable medium for the production of CA by the mold culture.Recipient of Deutscher Akademischer Austauschdienst (DAAD) fellowship 1984–1985 (National Dairy Research Institute, Karnal-132 001, India)     

Effects of air temperature and water vapor pressure deficit on storage of the predatory mite Neoseiulus californicus (Acari: Phytoseiidae)

To determine the optimum air temperature and water vapor pressure deficit (VPD) for the storage of the predatory mite, Neoseiulus californicus, 3-day-old mated females were stored at air temperatures of 0, 5, 10, or 15?°C and VPDs of 0.1, 0.3, or 0.5?kPa for 10, 20, or 30?days. At 10?°C and 0.1?kPa, 83?% of females survived after 30?days of storage; this percentage was the highest among all conditions. VPDs of 0.3 and 0.5?kPa regardless of air temperature, and an air temperature of 0?°C regardless of VPD were detrimental to the survival of the females during storage. Since the highest survival was observed at 10?°C and 0.1?kPa, the effect of the storage duration on the post-storage quality of the stored females and their progeny was investigated at 25?°C to evaluate the effectiveness of the storage condition. The oviposition ability of the stored females, hatchability, and sex ratio of their progeny were not affected even when the storage duration was extended to 30?days. Although a slight decrease in the survival during the immature stages of progeny was observed when the storage duration was ≥20?days, the population growth of N. californicus may not be affected when individuals stored in these conditions are applied to greenhouses and agricultural fields. The results indicate that mated N. californicus females can be stored at 10?°C and 0.1?kPa VPD for at least 30?days.     

Effect of temperature on the longevity of human fibroblasts in culture

The longevity of parallel cultures of the human diploid fibroblast strain MRC-5 was measured at various incubation temperatures. At 37°C the mean life-span was 57.2 passages, at 34°C it was 58.7 passages and at 40°C it was 29.2 passages. There was greater variation in longevity among cultures grown at 40°C than in the control population and least among those grown at 34°C. The decreased life-span at 40°C was probably due to accelerated ageing, as the transfer of senescent cultures back to 37°C did not lead to their recovery. Cultures grown at 32°C also had reduced life-span compared to the control, but this was probably not the result of ageing, as the transfer of cultures which had almost ceased growth back to 37°C allowed them to reach the normal life-span for this temperature. The results imply that clonal ageing is at least in part due to random events—possibly errors in protein synthesis—which occur more frequently with increasing temperature.     

Identification of Distinct Internal and External Isozymes of Carbonic Anhydrase in Chlorella saccharophila

External carbonic anhydrase (CA) was detected in whole cells of alkaline-grown Chlorella saccharophila but was suppressed by growth at acid pH or growth on elevated levels of CO2. Internal CA activity was measured potentiometrically as an increase in activity in cell extracts over that of intact cells. Cells grown under all conditions had equal levels of internal CA activity. Two isozymes were identified after electrophoretic separation of soluble proteins on cellulose acetate plates. The fast isozyme was found in cells grown under all conditions, whereas the slow isozyme was found only in cells grown at alkaline pH. Western blot analysis following sodium dodecyl sulfate-polyacrylamide gel electrophoresis using antibodies produced against the periplasmic form of CA from Chlamydomonas reinhardtii revealed a single band at 39 kD, which did not change in intensity between growth conditions and was associated only with proteins eluted from the fast band. The slow isozyme was inactivated by incubation of cell extract at 30[deg]C and by incubation in 10 mM dithiothreitol, whereas the internal form was unaffected. These results indicate that external and internal forms of CA differ in structure and their activities respond differently to environmental conditions.