Role of the competitive microbial flora in the radiation-induced enhancement of ochratoxin production by Aspergillus alutaceus var. alutaceus NRRL 3174.

The radiation sensitivity and the toxigenic potential of conidiospores of the fungus Aspergillus alutaceus var. alutaceus were determined after irradiation with 60Co gamma rays and high-energy electrons. Over the pH range of 3.6 to 8.8, the doses required for a 1 log10 reduction in viability based on the exponential portion of the survival curve ranged from 0.21 to 0.22 kGy, with extrapolation numbers (extrapolation of the exponential portion of the survival curve to zero dose) of 1.01 to 1.33, for electron irradiation, and from 0.24 to 0.27 kGy, with extrapolation numbers of 2.26 to 5.13, for gamma irradiation. Nonsterile barley that was inoculated with conidia of the fungus and then irradiated with either electrons or gamma rays and incubated for prolonged periods at 28 degrees C and at a moisture content of 25% produced less ochratoxin A with increasing doses of radiation. Inoculation of barley following irradiation resulted in enhanced ochratoxin levels compared with unirradiated controls. In these experiments, inoculation with 10(2) spores per g produced greater radiation-induced enhancement than inoculation with 10(5) spores per g. There was no radiation-induced enhancement when the barley was surface sterilized by chemical means prior to irradiation. These results are consistent with the hypothesis that a reduction in the competing microbial flora by irradiation is responsible for the enhanced mycotoxin production observed when nonsterile barley is inoculated with the toxigenic fungus A. alutaceus var. alutaceus after irradiation.     

Variants of Aspergillus alutaceus var. alutaceus (formerly Aspergillus ochraceus) with altered ochratoxin A production.

The present studies, using Aspergillus alutaceus var. alutaceus Berkeley et Curtis (formerly A. ochraceus Wilhelm) NRRL 3174 along with three other wild-type strains, were undertaken in an attempt to understand the effects of irradiation and other treatments on mycotoxin production in grain. Bedford barley was inoculated with spores of NRRL 3174, gamma irradiated, and incubated at 28 degrees C and 25% moisture. After 10 days of incubation, two colony types, ochre (parental) and yellow (variant), were isolated from the grain. Further culturing of the yellow variant resulted in the spontaneous appearance of a white variant that exhibited greatly enhanced fluorescence under UV light. In subsequent work, we have also isolated variants producing a soluble red pigment. In addition, in model experiments involving irradiation (1 kGy) of pure cultures, induction frequencies ranging between 2 and 4% (survival basis) were observed for the yellow and red variants. Inoculation of these variants into wheat and incubation for 14 days at 28 degrees C and 32% moisture resulted in ochratoxin A production in the relative amounts of 0.09:1:4.6:9.3 for the red, ochre (parental), yellow, and white variants, respectively. Additional characteristics of these isolates are described. Confirmation that the white high-ochratoxin-A-producing variants were derived from the parental strain was demonstrated by obtaining revertant sectors in monoclonal cultures of the variants.     

Variants of Aspergillus alutaceus var. alutaceus (formerly Aspergillus ochraceus) with altered ochratoxin A production.

The present studies, using Aspergillus alutaceus var. alutaceus Berkeley et Curtis (formerly A. ochraceus Wilhelm) NRRL 3174 along with three other wild-type strains, were undertaken in an attempt to understand the effects of irradiation and other treatments on mycotoxin production in grain. Bedford barley was inoculated with spores of NRRL 3174, gamma irradiated, and incubated at 28 degrees C and 25% moisture. After 10 days of incubation, two colony types, ochre (parental) and yellow (variant), were isolated from the grain. Further culturing of the yellow variant resulted in the spontaneous appearance of a white variant that exhibited greatly enhanced fluorescence under UV light. In subsequent work, we have also isolated variants producing a soluble red pigment. In addition, in model experiments involving irradiation (1 kGy) of pure cultures, induction frequencies ranging between 2 and 4% (survival basis) were observed for the yellow and red variants. Inoculation of these variants into wheat and incubation for 14 days at 28 degrees C and 32% moisture resulted in ochratoxin A production in the relative amounts of 0.09:1:4.6:9.3 for the red, ochre (parental), yellow, and white variants, respectively. Additional characteristics of these isolates are described. Confirmation that the white high-ochratoxin-A-producing variants were derived from the parental strain was demonstrated by obtaining revertant sectors in monoclonal cultures of the variants.     

Production of ochratoxin A by Aspergillus ochraceus NRRL-3174 before and after exposures to 60Co irradiation.

Spores from the toxigenic organism Aspergillus ochraceus NRRL-3174 were exposed to specific levels of gamma irradiation and then allowed to germinate on selected media. Increases in ochratoxin A production by irradiated, compared to non-irradiated, spores were observed after inoculation of spores onto a cracked red wheat or into a synthetic liquid medium. Variations in daily ochratoxin production were also observed for control and irradiated spore-derived cultures developing on both media, with maximum toxin production varying from 7 to 11 days of incubation. The most notable increases in ochratoxin A production occurred from cultures developing from spores having been irradiated with 10, 25, or 50 krad. Exposures to 400 or 600 krad resulted in complete inhibition of spore germination and, consequently, no ochratoxin production. Of the two substrates used, wheat and synthetic, the quantities of ochratoxin A produced were significantly lower in the synthetic media than on the natural substrate. Higher and more rapid toxin production occurred from spores having been irradiated with 10, 25, 50, and 100 krad than occurred from the non-irradiated control spores when grown on synthetic media. Cultures derived from spores having been exposed to 10, 25, 50, and 100 krad produced significantly higher levels of ochratoxin A after 8 days of incubation on natural substrate than did the controls. Analysis of variance revealed that substrate, length of incubation, as well as irradiation levels all affected the time required to produce maximum levels of ochratoxin A.     

Production of ochratoxin A in barley by Aspergillus ochraceus and Penicillium viridicatum: effect of fungal growth, time, temperature, and inoculum size.

Moistened barley was inoculated with 1.4 x 10(3) and 1.4 x 10(5) spores, respectively, from ochratoxin A-producing strains of Aspergillus ochraceus and Penicillium varidicatum. To estimate fungal tissue in the barley, the amount of glucosamine was followed for 28 days at 10 and 25 degrees C. Ochratoxin A was also followed during the same period and under the same conditions. The data show that ochratoxin A could be detected 4 to 6 days after inoculation at 25 degrees C, and the maximal accumulation of ochratoxin A was observed 28 days after inoculation. After 28 days at 25 degrees C, the quantities of ochratoxin A were between 7 and 46 micrograms/g of grain. At 10 degrees C only P. viridicatum produced ochratoxin A. The results indicated that production of ochratoxin A is not associated with rapid increase of glucosamine in the barley.     

Electron-beam sterilization of laboratory animal diets--sterilizing effect of 10-MeV electrons from a linear accelerator.

Electron beam sterilization for laboratory animal diets was examined as an alternative to 60Co gamma rays. Solid, powder diets for "mice and rats" and solid diets for "rabbits and guinea pigs" which are the main products sterilized by 60Co gamma rays were irradiated with 10-MeV electrons from a linear accelerator at the Research Institute for Advanced Science and Technology, Osaka Prefecture University. At least 20 kGy was required to sterilize the samples irrespective of solid or powder diets, which was in good accordance with the results for 60Co gamma rays. Using a set dose of 30 kGy, a thickness of 45 mm for solid diets and 30 mm for powder diets could be sterilized by "one-sided" irradiation. "Dual-sided" irradiation could sterilize all the solid diets and the powder diets contained in the thicknesses of 90 mm and 75 mm, respectively. Irradiation effects of 10-MeV electrons on the nutrient quality of each diet were almost equivalent to those of 60Co gamma rays. These results suggest that commercially adopted sterilization doses for 60Co gamma rays are applicable to electron sterilization without modification if the depth-dose profile and the minimum dose of irradiated samples are precisely assessed.     

Gamma radiation effects on growth and yield attributes of <Emphasis Type="Italic">Psoralea corylifolia</Emphasis> L. with reference to enhanced production of psoralen

The present work describes radiation-induced effects on vegetative, reproductive traits and psoralen content in Psoralea corylifolia L. The effects of gamma radiation on Psoralea seeds were investigated by exposing seeds with doses of 2.5, 5, 10, 15 and 20 kGy at dose rate of 1.65 kGyh⁻¹ and studying the plant growth at three developmental stages: preflowering, flowering and post flowering (seed to seed) after irradiation. Irradiation with lower doses of gamma rays significantly improved vegetative traits while higher doses proved depressing for same parameters. Similar trend was followed in reproductive traits. Psoralen, showed highest concentration in seeds (7.56%) at 20 kGy and lowest in control roots (0.23%). Increment in psoralen was striking for higher gamma doses applied. These long-term changes in plant development may be attributed to alteration in plant genome induced by irradiation. The results show in depth development stimulation and enhancement of secondary metabolite in Psoralea corylifolia L. following low and high dose treatment respectively depicting the potential of gamma rays in plant biotechnology and metabolomics.     

Gamma radiation resistance of Bacillus anthracis spores

The aim of the presented study was determined the effectiveness of action the gamma radiation on water suspension B. anthracis spores. The irradiation was performed using a Cobalt 60 (Co 60) source, by using single and fractionary irradiation doses. In the investigations was used B. anthracis stain "Sterne" 34F2. The obtained results show, that gamma radiation effectively inactivates B. anthracis spores. On the efficiency of sterilization process influence the irradiation's method and the number of spores in 1 ml suspension. In the suspension 1.5 x 10(9) spore in 1 ml, sporicidal doses gamma radiation amount to 25.0 kGy (single dose) or 41.5 kGy (fractionary dose). The volume suspension about definite inoculum of spores, subjected working the gamma rays has not influence on sporicidal effectiveness of radiation sterilization.     

Contribution of radiation-induced, nitric oxide-mediated bystander effect to radiation-induced adaptive response.

There has been a recent upsurge of interest in radiation-induced adaptive response and bystander effect, which are specific modes in stress response to low-dose/low-dose rate radiation. Recently, we found that the accumulation of iNOS in wtp53 celIs was induced by chronic irradiation with gamma rays followed by acute irradiation with X-rays, but not by each one, resulting in an increase in nitrite concentrations of medium. It is suggested that the accumulation of iNOS may be due to the depression of acute irradiation-induced p53 functions by pre-chronic irradiation. In addition, we found that the radiosensitivity of wtp53 cells against acute irradiation with X-rays was reduced after chronic irradiation with gamma rays. This reduction of radiosensitivity of wtp53 cells was nearly completely suppressed by the addition of NO scavenger, carboxy-PTIO to the medium. This reduction of radiosensitivity of wtp53 cells is just radiation-induced adaptive response, suggesting that NO-mediated bystander effect may considerably contribute to adaptive response induced by radiation.     

钴—60对高兔卵黄液中大肠杆菌的灭菌研究

用钴－60γ射线对接种于卵黄液中的大肠杆菌进行辐照灭菌,并测定了卵黄液的抗体效价、活性氧和卵黄液保存期的变化。结果表明：1.大肠杆菌在卵黄液中的D10值为0.31-0.37kGy,杀灭卵黄液中大肠杆菌的照射剂量为3kGy;2.辐照量在15kGy以下时,对卵黄液的抗体效价没有影响或仅有轻微影响;3.经8kGy照射后的卵黄液在常温、4℃、-10℃条件保存时,保存期明显长于未照射的卵黄液。     

The complexity of radiation-induced DNA damage as revealed by exposure to cell extracts.

The rejoining of single-strand breaks (SSBs) induced in plasmid DNA in the presence of 10 mmol dm(-3) Tris scavenger by aluminum K (Al(K)) ultrasoft X rays has been compared with that for SSBs induced by gamma radiation. The Al(K) ultrasoft X rays interact to produce low-energy secondary electrons, which are thought to be the main contributors to the formation of complex damage by low-LET radiations. The rejoining of radiation-induced SSBs was investigated using human whole cell extracts. The efficiency of rejoining of SSBs induced by Al(K) ultrasoft X rays is less than that observed for gamma-ray-induced SSBs. From the similarity of the extent of rejoining of SSBs induced by gamma rays under aerobic and anaerobic conditions, the chemical nature of the stand break termini does not significantly influence SSB rejoining. A simple nick induced in plasmid DNA by gpII protein is rejoined rapidly compared with the slower rejoining processes for radiation-induced SSBs. Therefore, ligation is not rate-determining in processing radiation-induced SSBs. This study provides further evidence that nonrejoining of radiation-induced SSBs reflects the complexity of DNA damage. From comparison of the extent of rejoining of SSBs induced by different radiations, it is inferred that double-strand breaks represent only a minor component of the overall yield of complex damage.     

The decay of potentially lethal oxygen-dependent damage in fully hydrated Bacillus megaterium spores exposed to pulsed electron irradiation.

Using a stopped-flow mixing and pulsed irradiation apparatus, a study has been made of the decay, to a harmless form, of radiation-induced species which would otherwise be lethal to spores on contact with oxygen. Aqueous suspensions of Bacillus megaterium spores were irradiated with 600 krad of electrons given over approximately 1 s; at various times after irradiation oxygen in solution was added. As the interval between irradiation and introduction of oxygen increased, the fraction of spores surviving increased. For spores irradiated in a deoxygenated condition the decay of the potentially lethal species, reflected by this change in survival, proceeded as if two parallel first-order reactions with half-lives of 9 and 120 s operate. In contrast, for spores equilibrated with oxygen and then irradiated, the decay is described by a single first-order expression with an associated half-life similar to that of the faster of the two reactions operating in anoxia.     

Effect of gamma sterilization on microhardness of the cortical bone tissue of bovine femur in presence of N-Acetyl-L-Cysteine free radical scavenger

Gamma sterilization is usually used to minimize the risk of infection transmission through bone allografts. However, it is believed that gamma irradiation affects the mechanical properties of allografts and free radical scavengers can be used to alleviate the radiation-induced degradation of these properties. The aim of this study was to investigate the radioprotective effects of N-Acetyl-L-Cysteine (NAC) free radical scavenger on the material properties of sterilized bovine cortical bone at microstructure level. Forty-two cortical tissue specimens were excised from three bovine femurs and irradiated to 35 and 70 kGy gamma rays in the presence of 5, 50, and 100 mM concentrations of NAC. The localized variations in microhardness were evaluated via indentation in the radial and longitudinal directions to examine different regions of the microstructures of the specimens, including the osteonal and interstitial tissues. A significant increase was observed in the hardness of osteonal, interstitial, and longitudinal combined microstructures exposed to 35 and 70 kGy radiations (P < 0.05), whereas a relative reduction of the hardness was observed in the radial direction. Furthermore, it was found that the application of 50 and 100 mM NAC during gamma irradiation significantly subsided the hardening in longitudinal combined microstructure. Moreover, the reduction of hardness in radial direction was suppressed in the presence of 100 mM of NAC. In conclusion, the results indicated that NAC free radical scavenger can protect the cortical bone against deteriorative effects of ionizing radiation and can be used to improve the material properties of sterilized allografts.     

Potential for biological control of Heterodera schachtii by Pochonia chlamydosporia var. chlamydosporia on sugar beet

The biological control potential of an isolate of Pochonia chlamydosporia var. chlamydosporia against Heterodera schachtii was examined by assessing the percentage of females and cysts that became infected on water agar, the effect of culture filtrate on juvenile mobility, and the effects of the fungus on the final population of the nematode on sugar beet under greenhouse conditions. After 3 weeks at 20°C, 74 and 95% of the eggs within cysts and females, respectively, were colonised by the fungus on water agar. The full concentration of the fungal filtrate from cultures in malt extract broth killed only 12% of the juveniles after 24 h at 25°C. In the greenhouse experiment, adding 16,000 chlamydospores of the fungus per gram of soil as either colonised barley grains or spores reduced the final number of females on roots of sugar beet by 50 and 66%, respectively, after 3 months. The reproduction factor was reduced to ×2 in spore-treated soil compared with ×5 in the untreated control, and 18% of the eggs in spore-treated soil were colonised by fungal mycelium. Generally, P. chlamydosporia var. chlamydosporia was more efficient at reducing the nematode population when applied as spores without any substrate than when used as colonised barley grains.     

DNA fragmentation by gamma radiation and electron beams using atomic force microscopy

Double-stranded pBS plasmid DNA was irradiated with gamma rays at doses ranging from 1 to 12 kGy and electron beams from 1 to 10 kGy. Fragment-size distributions were determined by direct visualization, using atomic force microscopy with nanometer-resolution operating in non-tapping mode, combined with an improved methodology. The fragment distributions from irradiation with gamma rays revealed discrete-like patterns at all doses, suggesting that these patterns are modulated by the base pair composition of the plasmid. Irradiation with electron beams, at very high dose rates, generated continuous distributions of highly shattered DNA fragments, similar to results at much lower dose rates found in the literature. Altogether, these results indicate that AFM could supplement traditional methods for high-resolution measurements of radiation damage to DNA, while providing new and relevant information.     

Sterilization of allograft bone: effects of gamma irradiation on allograft biology and biomechanics

Gamma irradiation from Cobalt 60 sources has been used to terminally sterilize bone allografts for many years. Gamma radiation adversely affects the mechanical and biological properties of bone allografts by degrading the collagen in bone matrix. Specifically, gamma rays split polypeptide chains. In wet specimens irradiation causes release of free radicals via radiolysis of water molecules that induces cross-linking reactions in collagen molecules. These effects are dose dependent and give rise to a dose-dependent decrease in mechanical properties of allograft bone when gamma dose is increased above 25 kGy for cortical bone or 60 kGy for cancellous bone. But at doses between 0 and 25 kGy (standard dose), a clear relationship between gamma dose and mechanical properties has yet to be established. In addition, the effects of gamma radiation on graft remodelling have not been intensively investigated. There is evidence that the activity of osteoclasts is reduced when they are cultured onto irradiated bone slices, that peroxidation of marrow fat increases apoptosis of osteoblasts; and that bacterial products remain after irradiation and induce inflammatory bone resorption following macrophage activation. These effects need considerably more investigation to establish their relevance to clinical outcomes. International consensus on an optimum dose of radiation has not been achieved due to a wide range of confounding variables and individual decisions by tissue banks. This has resulted in the application of doses ranging from 15 to 35 kGy. Here, we provide a critical review on the effects of gamma irradiation on the mechanical and biological properties of allograft bone.     

In vivo development of spores of Absidia ramosa.

Approximately 10(6) spores of Absidia ramosa were inoculated intravenously into normal and cortisone pretreated mice. At subsequent time intervals the liver, lungs and kidneys were removed and examined for fungal localization and growth. In normal mice, spore germination and continued hyphal growth was restricted to the kidneys-evidence of germination not being visible until around 30h post inoculation. Cortisone therapy allowed germination of spores in the lung and kidney by 7h but subsequent hyphal growth in the lung was severely restricted compared with the kidney where extensive hyphal growth occurred. Germination of spores in the liver of cortisone treated animals was slow, not becoming apparent until about 40h after inoculation. These results suggest that host defence mechanisms in the form of phagocytosis as well as biochemical inhibitors and/or lack of suitable stimulators are important in preventing germination of introduced fungal spores. Once germination has occurred, it appears that additional as yet undetermined factors play a role in allowing continued growth of the fungus.     

Sterilization of plastic containers using electron beam irradiation directed through the opening

AIMS: Studies were performed to demonstrate the efficacy of a novel electron beam irradiation system for the on-line sterilization of polymeric containers using energetic electrons directed through the container opening. METHODS AND RESULTS: The distribution of dosage delivered during conveyance beneath the electron beam treatment system was determined for two sizes (8 or 16 ounces) of high-density polyethylene (HDPE) blow-moulded bottles. The biological effects of treatment were then determined using Bacillus pumilus ATCC 27142 spores inoculated as 10 microl droplets dried (i) onto the surface of flat coupons of the high-density polyethylene bottle material, and (ii) onto the region of lowest delivered dose (the bottom side wall) within each bottle. The inactivation obtained was determined by examining the level of survival after swab recovery of the inoculated spores, with reference to the level recovered from untreated control samples. CONCLUSIONS: The inactivation of B. pumilus spores during treatment was reproducible and proportional to the applied dose. In each instance, the logarithm of the surviving fraction of spores was well fitted by linear regression models and in good general agreement with values reported for inactivation of the same organism using gamma irradiation. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this work demonstrate the potential of a new physical method capable of high throughput for in-line sterilization of polymeric containers. The ability of the process to eliminate the washing step involved with traditional chemical sterilants, such as hydrogen peroxide, greatly reduces the size and environmental impact of these systems.     

Enhanced symbiotic seed germination of Cypripedium macranthos var. rebunense following inoculation after cold treatment

Cypripedium macranthos var. rebunense is a well-known wild orchid in Japan, and is considered to be a symbol for rare plant conservation. A fungus isolated from roots of C. macranthos var. rebunense induced symbiotic germination of the species in vitro. Cold treatment of the seeds at 4°C prior to fungal inoculation was required for the symbiotic germination. Changing the timing of inoculation of the fungus to the seeds greatly improved germination frequency. Maximum germination was attained after seeds were inoculated just after the cold treatment for 12 weeks, and approximately 20% of the seeds developed into protocorms more than 1 mm long. These results suggest that fungal inoculation takes place at the beginning of spring in nature, and the tough impervious seed coat may preserve the seed from the infection during autumn and winter seasons. The lengthy culture period of more than 16 weeks at 20°C on the same medium with the fungus caused gradual browning and rot of the protocorms. By elimination of the fungus with a fungicide and by transfer to a nutrient rich medium, approximately 20% of the protocorms developed into healthy plantlets. The methods obtained here appear to be applicable to symbiotic germination of many other threatened Cypripedium spp.