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Biomedical subjects

O L Shotwell

Publications and source records attributed to O L Shotwell.

At least 19 recordsLinked to original sources
PubMed1988

Citreoviridin levels in Eupenicillium ochrosalmoneum-infested maize kernels at harvest.

Citreoviridin contents were measured in eight bulk samples of maize kernels collected from eight fields immediately following harvest in southern Georgia. Citreoviridin contamination in six of the bulk samples ranged from 19 to 2,790 micrograms/kg. In hand-picked samples the toxin was concentrated in a few kernels (pick-outs), the contents of which were stained a bright lemon yellow (range, 53,800 to 759,900 micrograms/kg). The citreoviridin-producing fungus Eupenicillium ochrosalmoneum Scott & Stolk was isolated from each of these pick-out kernels. Citreoviridin was not detected in bulk samples from two of the fields. Aflatoxins were also present in all of the bulk samples (total aflatoxin B1 and B2; range, 7 to 360 micrograms/kg), including those not containing citreoviridin. In Biotron-grown maize ears that were inoculated with E. ochrosalmoneum through a wound made with a toothpick, citreoviridin was concentrated primarily in the wounded and fungus-rotted kernels (range, 142,000 to 2,780,000 micrograms/kg). Samples of uninjured kernels immediately adjacent to the wounded kernel (first circle) had less than 4,000 micrograms of citreoviridin per kg, while the mean concentration of toxin in kernel samples representing the next row removed (second circle) and all remaining kernels from the ear was less than 45 micrograms/kg. Animal toxicosis has not been linked to citreoviridin-contaminated maize.

Animals↗
PubMed1984

Viable fungi in corn dust.

Numbers of viable fungal propagules in corn dusts in southern Georgia were estimated during various farm and grain elevator operations in 1979, 1980, and 1982. A six-stage Andersen sampler for viable microbial particles was used to sample the dusts with various agar media. The most abundant fungi in corn dusts were species of yeasts: Aspergillus, Penicillium, Cladosporium, Alternaria. Helminthosporium, and Fusarium. However, the relative abundance of these fungi differed between years. There was a greater incidence of the Aspergillus flavus group in the hot, dry year of 1980 compared with the cooler, wetter years of 1979 and 1982. Fungi in the corn dusts sampled numbered between 10(4) and 10(9) viable propagules per m3 of air. By contrast, outdoor air often contained fewer than 10(4) viable fungal propagules per m3. Most A. flavus propagules were deposited at stages three and four of the Andersen sampler, with correspond to the trachea, primary bronchi, and secondary bronchi in the human respiratory system. In an assessment of the air spores by exposing sterile petri dishes, more large-spored fungi, like Alternaria tenuis, and fewer small-spored fungi, such as A. flavus, were detected when compared with colony counts from petri dishes exposed to air in the Anderson sampler.

Air Microbiology↗
PubMed1983

Intrafungal distribution of aflatoxins among conidia and sclerotia of Aspergillus flavus and Aspergillus parasiticus.

This research examines the distribution of aflatoxins among conidia and sclerotia of toxigenic strains of Aspergillus flavus Link and Aspergillus parasiticus Speare cultured on Czapek agar (21 days, 28 degrees C). Total aflatoxin levels in conidia and sclerotia varied considerably both within (intrafungal) and among strains. Aspergillus flavus NRRL 6554 accumulated the highest levels of aflatoxin (conidia: B1, 84000 ppb; G1, 566000 ppb; sclerotia: B1, 135000 ppb; G1, 968000 ppb). Substantial aflatoxin levels in conidia could place at risk those agricultural workers exposed to dust containing large numbers of A. flavus conidia. Cellular ratios of aflatoxin B1 to aflatoxin G1 were nearly identical in conidia and sclerotia even though levels of total aflatoxins in these propagule types may have differed greatly. Aflatoxin G1 was detected in sclerotia of all A. flavus strains but in the conidia of only one strain. Each of the A. parasiticus strains examined accumulated aflatoxin G1 in both sclerotia and conidia. These results are examined in the context of current evolutionary theory predicting an increase in the chemical defense systems of fungal sclerotia, propagules critical to the survival of these organisms.

Aflatoxin B1↗
PubMed1983

Extraction and thin layer chromatography of aflatoxin B1 in mixed feeds.

A method was developed for the determination of aflatoxin B1 in commercially prepared feeds. The method incorporates methylene chloride and citric acid solution extraction, cleanup on a small silica gel column, and thin layer chromatography for quantitation. Commercial turkey starter, catfish chow, medicated pig starter, broiler finisher, rabbit chow, horse feed, rat chow, and dog chow were investigated. The feeds were spiked with naturally contaminated corn at 4 different levels of aflatoxin B1 (16-130 microgram/kg). Three assays were run on each of the 32 combinations of feed and levels of aflatoxin. Mean recoveries were 85.9-92.8% at levels of 16.5, 32.9, 65.8, and 131.6 micrograms/kg. The relative standard deviation per assay was 18.6%. This method is more rapid and less involved than most previously published methods for mixed feeds.

Aflatoxin B1↗
PubMed1983

Effect of feeding corn naturally contaminated with aflatoxin on feed efficiency, on physiologic, immunologic, and pathologic changes, and on tissue residues in steers.

Two of 3 groups of Holstein-Friesian steers (groups II and III; n = 5 each) were fed a ration containing corn naturally contaminated with 800 ng of aflatoxin/g. The other group of steers (group I; n = 5) was fed a ration containing noncontaminated corn. The respective rations were fed for 17.5 weeks, except the ration given to group III; the latter's first diet (contaminated with aflatoxin) was changed to a noncontaminated diet after 15 weeks, continuing for the remaining 2.5 weeks. All steers were killed and tissues and fluids were obtained for aflatoxin analysis. Although aflatoxin B1 and M1 could be detected in blood and urine at several sampling times during the experimental period in groups II and III steers (given the diets containing aflatoxin), there appeared to be no effects on body weight gains and immune phenomena, such as lymphoblastogenesis and antibody production, but there was a waning of the delayed cutaneous hypersensitivity in steers given aflatoxin-contaminated diets. In group III animals (diet was changed to noncontaminated ration at 15 weeks), aflatoxin B1 and M1 disappeared from urine before they were slaughtered. All tissues and fluids, except the rumen contents from these group III steers, were void of detectable aflatoxins B1 and M1 at necropsy. The concentrations of aflatoxin B1 in the rumen content of the latter steers were low. All tissues collected at necropsy from the group II steers fed the aflatoxin diet throughout the 17.5 weeks had detectable aflatoxins B1 or M1 present.

Aflatoxin B1↗
PubMed1983

Fate of aflatoxins in tissues, fluids, and excrements from cows dosed orally with aflatoxin B1.

A study was conducted to determine aflatoxins in tissues and non-tissues of 2 Holstein cows given oral doses of 0.35 mg of purified aflatoxin B1/kg of body weight/day for 3 consecutive days. Cow 1 was slaughtered 24 hours after the 3rd dose, and cow 2, after day 3, was fed aflatoxin-free rations for 7 additional days before slaughter. Tissue samples of brain, gallbladder and bile, heart, intestine, kidney, liver, lung, mammary gland, skeletal muscle, spleen, supramammary lymph nodes, thymus, and tongue, and nontissue samples of blood, feces, milk, rumen content, and urine were examined. Aflatoxins B1 and M1 were found in all samples of cow 1, except the thymus. Kidney, liver, and mammary gland had the highest concentrations of total aflatoxins (57.9, 13.2, and 25.1 ng/g, respectively), with the aflatoxin M1 concentration 40 times more than the aflatoxin B1 level in kidney. Aflatoxin residues were present (0.02 to 0.11 ng/g) only in kidney, liver, and intestine of the tissues from cow 2 (fed aflatoxin-free feed for 7 additional days). Aflatoxin B1 was not present in nontissue samples, but aflatoxin M1 (0.10 and 1.5 ng/ml) was found in the last milk and urine samples from the same cow. Urine assays are a possible way to monitor the presence of aflatoxin residues in meat tissues.

Aflatoxins↗
PubMed1983

Five-year study of mycotoxins in Virginia wheat and dent corn.

Every year during the 5-year period 1976-1980, approximately 100 samples each of corn and wheat from trucks delivering the grains at elevators in Virginia were collected by personnel of the Federal Grain Inspection Service and shipped to NRRC. Samples were analyzed as soon as possible for aflatoxin, zearalenone, and ochratoxin A. The 3 mycotoxins were not detected in any wheat sample. Zearalenone and ochratoxin A were not found in any corn sample; however, aflatoxin was detected in at least 25% of the corn samples from every crop year. In 1976-1980, the incidence of aflatoxin at levels of 20 ng/g or more (the Food and Drug Administration guideline) ranged from 18 to 61%; aflatoxin incidence above 100 ng/g was 5-29%. The average aflatoxin levels in corn samples collected in the 5 years varied from 21 to 137 ng/g. Moisture content of the samples was not determined, so aflatoxin levels given may be higher than they were at harvest. However, there are obviously differences from year to year. In freshly harvested corn samples collected by fieldmen of the Statistical Reporting Service in yield surveys in 1978 and 1979, aflatoxin incidence above the FDA guideline was 10 and 13%, and above 100 ng/g was 4 and 7%. The average aflatoxin level in all samples collected in 1978 was 13 ng/g and in 1979, 36 ng/g. Some aflatoxin can be expected yearly in Virginia corn, but the incidence and levels vary from year to year.

Food Contamination↗
PubMed1983

Gas chromatographic determination of deoxynivalenol in wheat.

Modifications to a published method are described for the determination of deoxynivalenol (DON) in wheat by gas chromatography with electron capture quantitation of the heptafluorobutyrate derivative. In the modified method, DON is extracted by shaking the sample with methanol-water on a wrist-action shaker, followed by filtration through rapid flow paper. One concentration step is eliminated, and a hexane wash is incorporated to remove toluene from the silica gel column. Recoveries of DON from wheat samples spiked at 0.1, 0.5, and 1.0 ppm ranged from 77.3 to 86.3% and averaged 81.5%.

Chromatography, Gas↗
PubMed1983

Treatment of freshly harvested 1980 Georgia dent corn samples collected for aflatoxin analysis.

In 1980, corn was harvested from six 15-ft rows in each of 67 fields in Georgia for aflatoxin analysis. Every sixth ear from each field was placed in a sample bag to be dried the day of collection. The rest of the corn was husked and shipped to Peoria in cardboard boxes. When undried ear samples arrived in Peoria, each sample was randomly separated into 5 equivalent subsamples. One set of 67 subsamples was shelled and dried as soon as possible to avoid further aflatoxin formation. Two other sets of 67 subsamples were stored 3 and 6 weeks before shelling and drying. The remaining 2 sets of ear samples were placed in plastic bags with 5% Monoprop (1 part propionic acid plus 1 part versite) and stored 3 and 6 weeks before shelling and drying. The samples dried in Georgia before shipping had an average total aflatoxin level of 217 ng/g. Samples shelled and dried immediately after arrival had an average level of 202 ng/g. Samples shelled and dried after 3 and 6 weeks of storage had average total aflatoxin levels of 417 and 387 ng/g, respectively. Samples stored 3 and 6 weeks in the presence of 5% Monoprop (2.5% propionic acid) had average total aflatoxin levels of 120 and 157 ng/g, respectively.

Aflatoxins↗
PubMed1981

Measurements of airborne aflatoxins during the handling of contaminated corn.

Sample of airborne dust generated the handling of aflatoxin-contaminated corn were collected and analyzed to assess potential exposures of farmers and other agricultural workers to these mycotoxins. Using high volume total dust samplers and a high volume Andersen sampler, downwind dust samples were collected on glass filter when contaminated corn was transferred by augers from a storage bin into a wagon and back into the storage bin. The aflatoxin B1 content of the 15 dust samples ranged from 12.5 to 204.3 ppb, with an average of 138 ppb; the aflatoxin B2 content ranged from 1.1 to 41.6 ppb, with an average of 24.6 ppb. The B1 and B2 levels of contamination in the bulk corn were 223.9 and 17.5 ppb, respectively. The gravimetric dust concentration in the air ranged from 7 mg/m3 to 417 mg/m3. The samples taken with an Andersen sampler indicate the dust is relatively coarse with only approximately 17% less than 7 micrometer. An analysis of the dust from each stage showed higher levels of aflatoxins in the larger first-stage particles than in the finer particles on the succeeding stages. The results of this study indicate that the dust generated when handling contaminated commodities also may be contaminated and represent a potential inhalation hazard. This fact, coupled with the extreme toxicity and carcinogenicity previously demonstrated in animal studies, suggests that appropriate measures be taken to prevent worker exposure during handling of contaminated materials.

Aflatoxins↗
PubMed1981

Use of aflatoxin-producing ability medium to distinguish aflatoxin-producing strains of Aspergillus flavus.

Aflatoxin-producing ability medium was tested for its ability to distinguish aflatoxin-positive from aflatoxin-negative strains of Aspergillus flavus in naturally occurring populations from corn at harvest. All of the aflatoxin-positive strains and some of the aflatoxin-negative strains produced aflatoxins when cultured on cracked corn. Although the data indicate that aflatoxin-producing ability medium is not entirely reliable in distinguishing potential aflatoxin-producing strains of A. flavus from nontoxigenic strains, it is significant that the medium did not yield false-positives.

Aflatoxin B1↗
PubMed1981

Thin layer chromatographic determination of aflatoxin in corn dust.

Methods adopted by the AOAC and the American Association of Cereal Chemists for determining aflatoxin in corn were modified, and techniques were developed for application to samples of less than 1 to 10 g instead of the specified 50 g samples. Analysis included chloroform extraction of dust samples or dust collected from glass fiber filters, purification of extracts on a silica gel column of appropriate size, and measurement of aflatoxin by either 1- or 2-dimensional thin layer chromatography (TLC). The solvent for 1-dimensional TLC was chloroform-acetone-water (91 + 9 + 1). Solvents for 2-dimensional TLC were, first direction, ether-methanol-water (95 + 4 + 1, lined tank) and second direction, chloroform-acetone-water (91 + 9 + 1, unlined tank), or first direction, chloroform-acetone-water (91 + 9 + 1, unlined tank) and second direction, toluene-ethyl acetate-formic acid (60 + 30 + 10, unlined tank). When samples weighed less than or equal to 0.1 g, the entire concentrated extract was applied to the TLC plate. About 0.5-1.0 ng aflatoxin B1 could be detected on the plate, making the limit of detection about 9 ng/g for 0.1 g samples.

Aflatoxin B1↗
PubMed1981

Minicolumn detection methods for aflatoxin in raw peanuts: collaborative study.

The Holaday-Velasco method and a modified Holaday method have been compared. The former method combines the speed and simplicity of the Holaday extraction and cleanup with the sensitivity of the minicolumn originally described by Velasco. The combination method has been approved by the AOAC and the AACC for determining aflatoxin in corn. The Holaday method was modified by substituting toluene for benzene in the solvent partition, and methylene chloride for chloroform in the minicolumn development to eliminate use of hazardous solvents. The neutral alumina in the Holaday minicolumn was changed from activity V to activity III to provide a more stable column. At aflatoxin levels in raw peanuts of 13-20 ng/g, the presence of aflatoxin was missed by the modified Holaday method in 4 analyses (3 laboratories) of 42 reported. There were no misses in this contamination range by the Holaday-Velasco method. There were no misses by either method with samples containing greater than 20 ng total aflatoxins/g. Analysis of uncontaminated raw peanuts by the modified Holaday method resulted in 2 false positives of 14 reports; the Holaday-Velasco method produced no false positive reports from 15 analyses of uncontaminated peanuts. The Holaday-Velasco method was adopted official first action for peanuts.

Aflatoxins↗
PubMed1981

Determination of aflatoxins in animal tissues.

A method for the determination of aflatoxins in animal tissues has been developed, and applied successfully to beef, swine, chicken, and human livers, and to beef kidney, heart, spleen, muscle, and blood. Blended tissue is denatured with citric acid and extracted with dichloromethane on a wrist-action shaker. After filtration, the extract is partially purified on a silica gel column, and aflatoxins B1 and M1 are determined by 2-dimensional thin layer chromatography and densitometry. Recoveries of B1 and M1 added to meat tissues and blood were approximately 90 and 80%, respectively. The method gave results for a contaminated freeze-dried liver comparable to analyses by 3 other published meat tissue methods. The method is rapid and has a determination limit less than or equal to 0.1 ng/g. In addition, the method uses less toxic and smaller quantities of solvents and chemicals.

Aflatoxins↗
PubMed1980

Laboratory screening for zearalenone formation in corn hybrids and inbreds.

Grains from 14 corn inbreds and 4 single cross hybrids were inoculated with 3 isolates of Gibberella zeae to determine their inhibition of zearalenone production. The corn hybrids: Pa762 x A632 (50 mg/kg zearalenone production), A619 x A632 (17 mg/kg zearalenone production), H95 x Mo17 (132 mg/kg zearalenone production), and B73 x MO17 (33 mg/kg zearalenone production) appear to have less resistance than the inbreds to toxin formation. Inbred H95 (64 mg/kg zearalenone production) supported the highest toxin production of all inbreds. The remaining 13 inbreds did not exceed 15 mg/kg zearalenone production. The inbreds A632 (4 mg/kg zearalenone production) and Pa762 (2 mg/kg zearalenone production) demonstrated some resistance; the resulting cross, hybrid Pa762 x A632 (50 mg/kg zearalenone production), does have greater resistance than hybrid H95 x Mo17 (132 mg/kg zearalenone production). Analysis of variance indicated highly significant variation between corn varieties and fungal isolates. The coefficient of variation for 29 fermentations run in duplicate on inoculated control corn to produce zearalenone (212 mg/kg) was 37%, which would include variation in both the fermentation and analysis. Isolate and variety interaction is not significant.

Fermentation↗
PubMed1980

Protocols for surveys, sampling, post-collection handling, and analysis of grain samples involved in mycotoxin problems.

This report examines and summarizes current knowledge regarding mycotoxin surveys, sampling techniques, conditions conducive to post-collection production of mycotoxins in grain samples, and analytical methods for mycotoxin analysis. Priority attention is given to samples of corn suspected of containing aflatoxin. The report includes recommendations where deems appropriate by the Ad Hoc Work Group.

Aflatoxins↗
PubMed1979

Minicolumn detection methods for aflatoxin in yellow corn: collaborative study.

The CPC modified method, the Holaday modified method, and the combination of the 2 procedures have been compared. The CPC modified method involves more cleanup steps but has a more sensitive column. The Holaday modified procedure has fewer cleanup steps, but the column is more difficult to interpret. The combination CPC-Holaday, which has proven to be the most satisfactory, combines the speed and simplicity of the Holaday extraction and the sensitivity of the Velasco minicolumn used in the CPC method. Levels of 10 ng/g were detected by 89% of the collaborating laboratories using the combination, Holaday-Velasco, method. The combination method has been adopted as official first action.

Aflatoxins↗