The effects of citrus oils and some spices on growth and aflatoxin production by Aspergillus parasiticus NRRL 2999

The inhibitory effect of various concentrations of mint, sage, bay leaves, thyme, aniseed, red pepper and citrus peel oils on the growth of an aflatoxin-producing strain of Aspergillus parasiticus and its production of aflatoxin was studied. Amongst the herbs used, thyme was found a highly effective antifungal agent. Growth and aflatoxin formation was depressed by orange and lemon oils at a concentration of 1.6% through 10 days incubation.     

Effect of Capsicum carotenoids on growth and aflatoxins production by Aspergillus flavus isolated from paprika and chilli

The aim of this study was to determine the effect of a carotenoid mixture (Capsantal FS-30-NT), containing capsanthin and capsorubin, on growth and aflatoxins (AF) production of AF-producing Aspergillus flavus isolates.     

Inhibitory effect of the essential oil of Curcuma longa L. and curcumin on aflatoxin production by Aspergillus flavus Link

Aflatoxins are highly toxic, mutagenic, teratogenic and carcinogenic mycotoxins. Consumption of aflatoxin-contaminated food and commodities poses serious hazards to the health of humans and animals. Turmeric, Curcuma longa L., is a native plant of Southeast Asia and has antimicrobial, antioxidant and antifungal properties. This paper reports the antiaflatoxigenic activities of the essential oil of C. longa and curcumin. The medium tests were prepared with the oil of C. longa, and the curcumin standard at concentrations varied from 0.01% to 5.0%. All doses of the essential oil of the plant and the curcumin standard interfered with mycotoxin production. Both the essential oil and curcumin significantly inhibited the production of aflatoxins; the 0.5% level had a greater than 96% inhibitory effect. The levels of aflatoxin B₁ (AFB₁) production were 1.0 and 42.7 μg/mL, respectively, for the samples treated with the essential oil of C. longa L. and curcumin at a concentration of 0.5%.     

Inhibition of microbial growth at different pH levels by benzoic and propionic acids and esters of p-hydroxybenzoic acid

The minimum growth inhibitory concentration of benzoic and propionic acids and esters of p-hydroxybenzoic acid (parabens) has been determined for Bacillus subtilis, Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. For benzoic and propionic acid, the inhibition was shown to be due to both undissociated and dissociated acid, and the effect of each has been calculated in accordance with a proposed mathematical model which has earlier been applied to sorbic acid. Parabens showed some pH dependence for the action on bacteria, but not on yeast. This pH dependence was unrelated to dissociation.     

Combined effect of temperature and propionic acid concentration on the growth of Aspergillus parasiticus

Effects of temperature (25, 30 and 36°C) and propionic acid concentration (129, 258 and 516 ppm) on the growth of Aspergillus parasiticus in solid media were analyzed. Growth rate, expressed as the increment of colony diameter per unit of time, was studied as a function of storage time employing the Arrhenius model. The inverse of the lag phase was fitted to an Arrhenius-type equation and the inhibition index was also calculated. A linear relationship between the lag phase and the reciprocal growth rate at different propionic acid concentrations was assessed by linear regression analysis. Fungi behavior was modeled considering the main controlling factors and a response surface methodology was established in terms of propionic acid and temperature. The proposed model could be used in food microbiology to predict the growth of toxicogenic fungi growth.     

Effect of application of nontoxigenic strains of Aspergillus flavus and A. parasiticus on subsequent aflatoxin contamination of peanuts in storage

Experiments were conducted to determine the potential for biological control of aflatoxin contamination of peanuts during storage. Florunner peanuts were treated in field plots by applying competitive, nontoxigenic strains of Aspergillus flavus and A. parasiticus, at 76 and 67 days after planting in 1998 and 1999, respectively. After harvest, half the peanuts from both treated and control plots were sprayed with an aqueous conidial suspension containing the nontoxigenic strains; the other half of the peanuts from each group were not sprayed. The peanuts were then placed in separate compartments of a miniature warehouse. Therefore, storage treatments consisted of peanuts that were (1) not treated at all; (2) treated prior to storage only; (3) field-treated only; (4) treated both in the field and prior to storage. Peanuts were stored for 3-5 months under high temperature and relative humidity conditions designed to promote aflatoxin contamination. In 1998, peanuts were not contaminated with aflatoxins prior to storage. After storage, peanuts that were never treated with the competitive fungi contained an average of 78.0 ppb of aflatoxins. Peanuts not treated in the field but receiving the spray treatment before storage contained 48.8 ppb. Peanuts treated in the field only averaged 1.4 ppb, and peanuts treated both in the field and prior to storage contained 0.8 ppb. In 1999, peanuts suffered from late-season drought and were contaminated with aflatoxins at harvest, with controls averaging 516.8 ppb compared with 54.1 ppb in treated peanuts. After storage, non-field-treated peanuts averaged 9145.1 ppb compared with 374.2 ppb for peanuts that had been field-treated, a 95.9% reduction. Spraying of pods with the nontoxigenic strains postharvest but prior to storage provided no additional protection against aflatoxin contamination. Results demonstrated that field application of the nontoxigenic strains had a carry-over effect and reduced aflatoxin contamination that occurred in storage.     

Activity of essential oil and its major compound, 1,8-cineole, from Eucalyptus globulus Labill., against the storage fungi Aspergillus flavus Link and Aspergillus parasiticus Speare

The essential oil from leaves of Eucalyptus globulus obtained by hydrodistillation, as well as its major compound 1,8-cineole, identified by gas chromatography coupled with a mass selective detector, were evaluated for their effectiveness against the storage fungi Aspergillus flavus and Aspergillus parasiticus. The evaluation was performed by compound dissolution in yeast extract sucrose (YES) medium and exposure to headspace volatiles. Complete fungal growth inhibition of both species was achieved with the essential oil by contact and volatile assays. Volatile exposure showed total inhibition at the lower level tested of 500 μL. The 1,8-cineole tested alone showed partial inhibition only at the highest level of 1.3492 μL. Aflatoxin B₁ production was reduced in headspace volatile assays and partial inhibition was observed at the 200 μL dose of the essential oil.     

Effect of gas barrier characteristics of films on aflatoxin production by Aspergillus flavus in peanuts packaged under modified atmosphere packaging (MAP) conditions

The effects of the gas barrier characteristics of three films (ASI, ASII and ASIII) and storage temperature on the growth of, and aflatoxin production by, Aspergillus flavus in peanuts packaged in air and under a modified gas atmosphere of CO₂:N₂ (65:35) were investigated. Mold growth was barely visible in air packaged peanuts using high-medium barrier films (ASI and ASII) and stored at 20°C with more extensive growth occurring in air packaged peanuts stored at 25 and 30°C. Extensive mycelial growth and sporulation occurred in all air packaged peanuts in a low barrier film (ASIII), especially at 30°C. Gas packaging inhibited mold growth in peanuts packaged in a high gas barrier film at 20°C. However, mold growth occurred in gas packaged peanuts packaged in film ASII at higher storage temperatures while extensive mycelial growth was observed in all peanuts packaged in film ASIII irrespective of storage temperature. Levels of aflatoxin greater than the regulatory limit of 20 ng/g were detected in all air packaged peanuts with the highest level of aflatoxin (76 ng/g) being detected in peanuts packaged in a high gas barrier film ASI. Aflatoxin production was inhibited in gas packaged peanuts using a high barrier film. However, higher levels of aflatoxin were detected in all gas packaged peanuts in medium-low gas barrier films (ASII and ASIII), particularly at higher storage temperatures. This study has shown that MAP using a CO₂:N₂ (65:35) gas mixture was effective in controlling aflatoxin production by A. flavus in peanuts to levels less than the regulatory limit of 20 ng/g. However, the antimycotic effect of low O₂-high CO₂ atmospheres is dependent on the gas barrier characteristics of the packaging films, especially at higher storage temperatures.     

Effect of dietary fiber and butyric acid on lysosomal enzyme activities in streptozotocin-induced diabetic rats

The effect of dietary fiber and butyric acid on lysosomal enzyme activities in different tissues of streptozotocin-induced diabetic rats was studied by analyzing the specific activities of N-acetyl-β-d-glucosaminidase, β-d-glucuronidase, β-d-galactosidase, and acid phosphatase in various tissues. An alteration in the activities of these lysosomal enzymes in different tissues was observed in streptozotocin-induced diabetic rats. Dietary fiber and butyric acid supplementation at 500 mg/kg body weight/day was found effective in modulating the altered glycosidases activity in liver, spleen, heart, lungs, testis, and brain during diabetes. Our results also indicated that butyric acid along with dietary fiber is involved in minimizing the oxidative stress-induced tissue damage thereby regulating the altered enzyme activities in different tissues during diabetes.     

Degradation of cereal bran polysaccharide-phenolic acid complexes by Aspergillus niger CFR 1105

Cereal brans such as wheat and rice are abundant sources for obtaining bioactive phenolic compounds such as ferulic and coumaric acids, in turn these can be bio-transformed into high value flavour compounds such as vanillin. Aspergillus niger CFR 1105, found to induce greater amounts of cell wall degrading enzymes, was inoculated into wheat and rice brans, and grown for 24, 48, 72 and 96 h to understand the degradation pattern of non-starch polysaccharides and phenolic acid complexes. Native wheat bran polysaccharides mainly consisted of arabinose, xylose, galactose, and glucose, in % of 27:39:2:30, with traces of mannose, whereas rice bran consisted of arabinose, xylose, galactose, and glucose in % of 9:27:31:32, respectively. Both the cereal bran arabinoxylans were degraded extensively by A. niger at 96 h, whereas the degradation of 1,3/1,4-β-d-glucans and cellulose was negligible. Bound phenolic acids of rice and wheat brans identified by HPLC were found to be mainly ferulic, coumaric, syringic acids in % of 93.6:6.3:0 (wheat), 34.7:55.8:9.5 (rice) and were drastically degraded/utilized at 96 h. The above results have indicated preferential degradation of arabinoxylans of wheat bran by A. niger, which can be exploited to obtain bioactive compounds such as ferulic acid.     

Effect of cell immobilization on heat-induced sublethal injury of Escherichia coli, Salmonella Typhimurium and Listeria innocua

The occurrence of sublethally injured cells in foods poses major public health concerns and is an essential aspect when assessing the microbial response to food preservation strategies, yet there is limited research dealing with its specific implications for mild heating. All available studies so far have been performed in broths colonized by planktonic cells, although their susceptibility to lethal agents has often been reported to be markedly different to the stress tolerance of cell colonies developed in solid foods. In this work, the effect of planktonic and colony growth, as well as the influence of colony density on sublethal injury induced by mild heating of Escherichia coli, Salmonella Typhimurium and Listeria innocua were assessed in food model systems. Detection of injured survivors relied on their inability to form visible colonies on salt-based selective media, which do not affect the growth of healthy cells. Sublethal injury (SI) increased rapidly with shorter exposure times and afterwards, decreased progressively, suggesting a mechanism of cumulative damage triggering lethal instead of SI. Cell arrangement affected the degree of SI, higher values being generally found for gelified systems, although the effect of colony density depended on the target microorganism. This information is essential for optimizing the design of food safety assurance systems.     

Effect of inoculum level on aflatoxin production by Aspergillus flavus under modified atmosphere packaging (MAP) conditions

The combined effect of inoculum level and other environmental factors (water activity, storage temperature and level of headspace oxygen) on the growth and aflatoxin production by Aspergillus flavus under MAP conditions on a synthetic medium was studied using a full factorial experimental design. Regression analysis of the data showed that inoculum level, water activity (a_w), storage temperature and headspace O₂ all had a significant effect on both growth of, and aflatoxin production by, A. flavus. Maximum growth (4·17 mg g^-1) occurred at an inoculum level of 10⁴ spores per plate, an a_w of 0·95, storage temperature of 20°C and headspace O₂ of 15% (balance CO₂:N₂ (60%:40%)). However, maximum aflatoxin level of 3·45 μg g^-1 (3450 ng g^-1) occurred at an inoculum level of 10³ spores per plate, an a_w of 0·96, storage temperature of 25°C and a reduced level of headspace O₂ (10%). Packing under almost completely anaerobic conditions using Ageless oxygen absorbents inhibited both growth and aflatoxin production by A. flavus, irrespective of the inoculum levels and storage temperature used.     

Effect of anilinopyrimidine resistance on aflatoxin production and fitness parameters in Aspergillus parasiticus Speare

Mutants of Aspergillus parasiticus resistant to the anilinopyrimidine fungicides were isolated at a high mutation frequency after UV-mutagenesis and selection on media containing cyprodinil. In vitro fungitoxicity tests resulted in the identification of two predominant resistant phenotypes that were highly (R₁-phenotype) and moderately (R₂-phenotype) resistant to the anilinopyrimidines cyprodinil, pyrimethanil and mepanipyrim. Cross-resistance studies with fungicides from other chemical groups showed that the highly resistance mutation(s) did not affect the sensitivity of R₁-mutant strains to fungicides affecting other cellular pathways. Contrary to that, a reduction in the sensitivity to the triazoles epoxiconazole and flusilazole, the benzimidazole carbendazim, the phenylpyrrole fludioxonil, the dicarboximide iprodione and to the strobilurin-type fungicide pyraclostrobin was observed in R₂-mutant strains. Study of fitness parameters of anilinopyrimidine-resistant strains of both phenotypic classes showed that all R₁ mutant strains had mycelial growth rate, sporulation and conidial germination similar to or even higher than the wild-type parent strain, while these fitness parameters were negatively affected in R₂ mutant strains. Analysis of the aflatoxin production showed that most R₁ mutant strains produced aflatoxins at concentrations markedly higher than the wild-type parent strain. A considerable reduction in the aflatoxin production was observed on cultured medium and on wheat grains by all R₂ mutant strains, indicating a possible correlation between fitness penalties and aflatoxigenic ability of A. parasiticus. The potential risk of increased aflatoxin contamination of agricultural products and their byproducts by the appearance and predominance of highly aflatoxigenic mutant strains of A. parasiticus resistant to the anilinopyrimidines is discussed.     

Effects of copper on germination,growth and sporulation of Clostridium tyrobutyricum

The effects of copper (Cu²⁺) on spore germination, vegetative growth and sporulation of Clostridium tyrobutyricum, which is capable to causing texture and flavour defects in Emmental cheese, were studied. Spore suspensions of three different strains were used as starting material for two experimental set-ups. The first studied the effects of supplemented (0–30 ppm) copper in RCM medium during spore germination and vegetative growth of C. tyrobutyricum measured by plating. The second set-up studied the effects of copper (0–30 ppm) in RCM medium during growth and sporulation of C. tyrobutyricum as measured by optical density at 550 nm and by platings after heat treatment of the samples respectively. Inhibition of germination, vegetative growth and sporulation processes by copper was strain-dependent. Both sporulation and germination were more sensitive than vegetative growth of C. tyrobutyricum to the inhibitory effects of copper. Copper, at the concentrations investigated in this study, inhibits spore germination of C. tyrobutyricum strains. Consequently copper may reduce the risk of late blowing spoilage from in the germination of C. tyrobutyricum spores during the ripening period of Emmental cheese.     

Effect of γ‐irradiation on aflatoxin B1 production by Aspergillus flavus and chemical composition of three crop seeds

The effect of γ‐irradiation on aflatoxin B₁ production by Aspergillus flavus, and the chemical composition of some different crop seeds were investigated. A. flavus infected seeds behaved differently according to their principal constituents. A. flavus caused an increase in protein and decrease in lipids and carbohydrate contents of wheat, soyabean and fababean seeds. Growth of A. flavus and production of aflatoxin B₁ was inhibited at a dose level of 5 kGy. A. flavus utilizes carbohydrates of seeds for its growth and aflatoxin production. Crops were arranged, in descending order, according to aflatoxin produced in seeds as wheat > soyabean > fababean. There were no changes in chemical constituents of irradiated seeds, such as protein, lipids, and carbohydrates.     

氮浓度对异养小球藻生长及蛋白质含量的影响

为了提高异养小球藻蛋白质含量低的问题,研究了异养小球藻生长及蛋白质含量和氮浓度的关系,探究了分阶段调控氮浓度对异养小球藻生长及蛋白质含量的影响。结果表明,氮浓度在3～15mmol/L范围内,小球藻生物量及蛋白质含量随氮浓度增加而增加,生物量从0.91g/L提高到了3.02g/L,蛋白质含量从26.1%提高到了37.4%。分阶段培养小球藻,首先在低氮条件下培养至指数期前期,然后转移至高氮浓度下培养,生物量达3.04g/L,且蛋白质含量提高至53.8%,与自养培养条件下蛋白质含量相当。     

Short communication: Effect of grazing on the concentrations of total sialic acid and hexose in bovine milk

Sialic acid, which is located at the terminal end of glycoconjugates, is believed to have important biological functions. Its concentration in bovine milk varies depending on lactation stage and season. However, it remains unclear whether dietary factors, especially fresh forage, affect the total sialic acid concentration in milk. The purpose of the present study was to investigate the effect of grazing on the concentrations of total sialic acid and hexose in bovine milk. Six healthy dairy cows were used in a crossover design (3 cows fed fresh forage and 3 cows fed grass silage) for 2 wk. Individual milk samples were collected at 2 consecutive milkings (morning and evening) at 0, 1, 3, 5, 8, 11, and 14 d of the experimental period, and 2 consecutive samples in each cow were combined on each sampling day in proportion of the morning and evening milk yields. No differences in body weight, milk yield, or milk composition were observed between the 2 groups during the experimental period. The hexose concentration in milk did not differ between these groups during the experimental period. Conversely, the total sialic acid concentration in the milk of each grazing cow significantly increased at 11 and 14 d of the experimental period compared with that at 0 d. In the grass silage group, the total sialic acid concentration at the end of the experimental period tended to be lower than that at 0 d, but the decrease was not significant. These results indicate that grazing management could have increased the concentration of sialoglycoconjugates in milk. This suggests that grazing may increase the biological function of milk because it is thought that sialic acid is significant in many ways.     

Effects of food preservatives on Alternaria alternata growth and tenuazonic acid production

The effects of different organic acids on Alternaria alternata growth and tenuazonic acid production (TeA) were evaluated. Both TeA pure toxin solution and TeA production in solid medium were considered. Sodium benzoate, potassium sorbate and sodium propionate, all preservatives commonly used by food industry in Argentina, were tested. TeA was stable as pure toxin solution when was treated with the salts of organic acids used. A differential effect was observed when the preservatives were evaluated in relation to A. alternata growth and TeA production in solid medium. L evels above 10mg/kg of sodium benzoate and potassium sorbate produced a total inhibition of fungal development and toxin biosynthesis. Sodium propionate produced a 59% decrease in A. alternata growth and total inhibition of TeA production only at the highest concentration of preservatives used.