Production of patulin and citrinin by Penicillium expansum from British Columbia (Canada) apples.

Twenty-four isolates of Penicillium expansum Link from British Columbia (Canada) apples were cultured in yeast-extract sucrose (YES) at 25°C for 28 days to investigate production of patulin and citrinin. These isolates proved to be potent producers of citrinin, patulin, or in most cases, both mycotoxins. In every isolate, citrinin, patulin, or both compounds were produced at levels as high as 565 µg/mL (mean 269 µg/mL) and 100 µg/mL (mean 31 µg/mL), respectively. Of the 24 isolates, 4 produced citrinin only, and 2 produced patulin only. Overall, 83% of the isolates formed patulin and 91% formed citrinin. YES broth proved to be an effective medium for patulin and citrinin production. Other workers have noted that production of these mycotoxins in culture often presages production in fruits, so these results might help Canadian fruit processors evaluate and minimize mycotoxin levels in their products.

Comparison of Canadian Fusarium graminearum isolates for aggressiveness, vegetative compatibility, and production of ergosterol and mycotoxins.

Fusarium graminearum is the predominant pathogen causing fusarium head blight of cereals in North America. Fifteen Canadian isolates of Fusarium graminearum were highly diverse in terms of vegetative compatibility grouping (VCG) and varied for production of ergosterol and mycotoxin production in rice culture. Aggressiveness was assessed by scoring the disease severity incited in wheat spikes by each isolate. Two inoculation methods, single-floret injection and spray of entire spikes, were used to screen 4 wheat varieties for reaction to the F. graminearum isolates. All isolates were of broadly similar aggressiveness, with disease severity ranging from 17.2 to 39.1 for single floret injection, and 39.1 to 69.0 for spray inoculation. Disease severity, ergosterol production, and mycotoxin development were not correlated. Using nitrate non-utilizing mutants the 15 isolates were grouped into 14 VCGs. Deoxynivalenol (DON) was produced by all isolates in rice culture, at levels between 0.2 and 249 ppm. 15-acetyldeoxynivalenol was produced by 14 of the 15 isolates at levels between 0.4 and 44.6 ppm. These results reveal a high level of diversity for several characteristics among F. graminearum isolates from Canada.

Trichothecene and moniliformin production by Fusarium species from western Canadian wheat.

Fusarium graminearum, Fusarium culmorum, and Fusarium avenaceum, isolated from Fusarium-damaged wheat harvested in western Canada, were cultured and evaluated for mycotoxin production. Extracts of the culture media were assayed for trichothecenes by gas chromatography-mass spectrometry and for moniliformin by liquid chromatography. Deoxynivalenol (DON) was found in 28 of 42 isolates of F. graminearum and 42 of 42 isolates of F. culmorum at levels ranging from 0.5 to 25.0 microg/g. 15-AcetylDON was found in 28 of 42 isolates of F. graminearum at levels ranging from 1.0 to 7.1 microg/g. 3-AcetylDON was found in 41 of 42 isolates of F. culmorum at levels ranging from 0.8 to 13.0 microg/g. Several other trichothecenes were assayed but not detected in the culture medium. Moniliformin was present in 40 of 42 isolates of F. avenaceum at levels ranging from 1.3 to 138.1 microg/g, but was not present in any of the isolates of F. graminearum or F. culmorum.

Relationships among deoxynivalenol, ergosterol and Fusarium exoantigens in Canadian hard and soft wheat.

Soluble extracellular components (exoantigens) from cultures of Fusarium graminearum and F. sporotrichioides were used to produce antisera from chickens for an indirect enzyme immunoassay. This immunoassay was used to estimate Fusarium exoantigen levels in 40 samples of fusarium head blight-infected hard red spring wheat from Manitoba, and in 50 samples of infected soft white winter wheat from Ontario. These wheat samples were also assayed for deoxynivalenol (DON), the predominant Fusarium mycotoxin, and for ergosterol, a metabolite reflecting fungal biomass. Using F. sporotrichioides antisera, the linear correlations between exoantigen level and DON content for the hard and soft wheats had coefficients of 0.80 and 0.76, respectively. With the same antisera, linear correlations between exoantigen level and total ergosterol concentration for the hard and soft wheats had coefficients of 0.66 and 0.81, respectively.

The characterization of chicken antibodies raised against Fusarium spp. by enzyme-linked immunosorbent assay and immunoblotting.

Immunoassays for the detection of three Fusarium species have been developed. Chickens were immunized with the soluble antigens from within the mycelia (mycelia-soluble antigens) and the exoantigens of Fusarium sporotrichioides, Fusarium poae and Fusarium graminearum and the antigens were characterized using enzyme-linked immunosorbent assays, immunoblotting and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The cross-reactivities of antisera with eleven species of Fusarium, eleven species of fungi from other genera and the buffer-extracts of grain were determined. Antisera to the exoantigens and mycelia-soluble antigens did not cross-react with buffer-extracts of grain. The antisera against the mycelia-soluble antigens tended to cross-react with the antigens from other genera whereas the antisera against the exoantigens were genus specific. The antiserum raised against exoantigens from F. poae was species specific. The molecular weights of the immunodominant antigens from the Fusaria were above 28 kilodaltons. Antigens from Fusaria showed specific bands in SDS-PAGE gel that can be used to produce more specific antibodies. The results suggest that exoantigen immunoassay can be developed to identify Fusarium genus, whereas mycelia-soluble antigens are not suitable for immunoidentification among genus.

Natural contamination of Manitoba barley by 3,15-diacetyldeoxynivalenol and its detection by immunochromatography.

Contamination of Canadian barley samples by 3,15-diacetyldeoxynivalenol was detected by enzyme immunoassays combined with liquid chromatography-mass spectrometry. This is the first reported natural occurrence of this mycotoxin. The barley was infected mainly with Fusarium graminearum. Deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone were also found.

A Convenient Method for Assessing Mycotoxin Production in Cultures of Aspergilli and Penicillia †.

Production of aflatoxins, sterigmatocystin, citrinin, and ochratoxin A by Aspergillus and Penicillium species on agar in 50-ml flasks after 21 days was detected and confirmed by thin-layer chromatography. Toxins were extracted with 2.5 ml of acidified methylene chloride for 20 to 60 min. Mycotoxin concentrations were estimated by liquid chromatography. Addition of formic acid to the methylene chloride extraction solvent effected an increase in recovery of ochratoxin A from Czapek-yeast (CY) agar from 20% to 91%. Recoveries of added mycotoxins were generally higher from CY agar than from yeast extract-sucrose (YES) agar and ranged from 11% (citrinin from YES agar) to 91% (ochratoxin A from CY agar). The method described is convenient, rapid, and reproducible, since it samples the entire culture surface. Human exposure to volatile solvents is also minimized.

Effects of tebufelone (NE-11740), a new anti-inflammatory drug, on arachidonic acid metabolism.

Tebufelone is a novel nonsteroidal anti-inflammatory drug (NSAID), of the di-tert-butylphenol (DTBP) class, which displays potent anti-inflammatory, analgesic and anti-pyretic properties in a variety of animal models. In this report, the effects of Tebufelone on arachidonic acid (AA) metabolism are reviewed. Tebufelone potently inhibits the formation of prostaglandins (PGE2) a key mediator of pain and inflammation, in isolated enzyme preparations (IC50 = 1.5 microM, KI = 0.35 microM), two in vitro cellular systems: rat peritoneal macrophages (IC50 = 0.02 microM) and human whole blood (IC50 = 0.08 microM), and ex vivo in man. In addition to PGE2 inhibition, which is common to all NSAIDs, higher concentrations of Tebufelone block the in vitro formation of products of the lipoxygenase pathway [leukotrienes (LTB4)] in rat macrophages (IC50 = 20 microM) and human whole blood (IC50 = 22 microM). Substrate incorporation studies (14C-AA) indicate that Tebufelone reversibly inhibits cyclooxygenase (CO) and 5-lipoxygenase (5-LO) enzymes rather than regulating the release of AA. Tebufelone was shown to be a more potent CO inhibitor than indomethacin and a less potent 5-LO inhibitor than RG-5901. Comparisons to structurally related compounds under development (E-5110, Esai; KME-4, Kanagafuchi), found Tebufelone to be the most potent CO inhibitor in vitro. All three DTBP compounds were equipotent 5-LO inhibitors. It is likely that Tebufelone's inhibitory effects on AA metabolism are, in part, responsible for its in vivo efficacy and enhanced safety profile.

Human electroretinogram responses to video displays, fluorescent lighting, and other high frequency sources.

Time-averaged human electroretinogram (ERG) responses were determined for several workplace visual stimuli which are temporally modulated at rates exceeding the perceptual critical fusion frequency (CFF). A clearly identifiable synchronous response was in evidence for a video display terminal (VDT) stimulus operating with a refresh rate as high as 76 Hz. A directly viewed fluorescent luminaire with controllable driving frequency elicited a synchronous response at rates as high as 145 Hz. In addition, an intense stimulus created by modulating the light from a slide projector produced responses at least as high as 162 Hz. The implications of these high-frequency responses are representing a potential basis for visual symptoms are discussed.

Computerized tomography of combined carcinomas arising in pleomorphic adenoma.

A case of carcinoma ex pleomorphic adenoma of the palate and maxillary sinus is reported. Unlike most such cases, there were two distinct malignant components: adenoid cystic carcinoma and adenocarcinoma. The computerized tomography of this lesion is described.