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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.

Aflatoxins↗

Step of dichlorvos inhibition in the pathway of aflatoxin biosynthesis.

Dichlorvos (dimethyl 2,2-dichlorovinyl phosphate) inhibits the biosynthesis of aflatoxin by Aspergillus parasiticus. Cultures treated with dichlorvos excrete an orange pigment which can be converted into aflatoxin B(1) by the untreated mycelia. The orange pigment was partially identified as an acetyl derivative of versiconol-type compound. In the presence of dichlorvos, sterigmatocystin is converted into aflatoxin B(1) without being interfered, but averufin is converted into the orange pigment instead of aflatoxin B(1). Therefore, dichlorvos appears to block an enzymatic step in the aflatoxin biosynthetic pathway, which lies beyond averufin but before sterigmatocystin, at the formation of the orange pigment.

Acetates↗

Mechanisms of aflatoxin carcinogenesis.

Much progress has been made in elucidating the biochemical and molecular mechanisms that underlie aflatoxin carcinogenesis. In humans, biotransformation of AFB1 to the putative carcinogenic intermediate. AFB-8,9-exo-epoxide, occurs predominantly by cytochromes P450 1A2 and 3A4, with the relative importance of each dependent upon the relative magnitude of expression of the respective enzymes in liver. Genetic variability in the expression of these and other cytochromes P450 may result in substantial interindividual differences in susceptibility to the carcinogenic effects of aflatoxins. Detoxification of AFB-8,9-epoxide by a specific alpha class glutathione S-transferase is an important protective mechanism in mice, and it accounts for the resistance of this species to the carcinogenic effects of AFB. This particular form of GST is expressed constitutively only at low levels in rats, but it is inducible by antioxidants such as ethoxyquin, and it accounts for much of the chemoprotective effects of a variety of substances, including natural dietary components that putatively act via an "antioxidant response element" (ARE). In humans, the constitutively expressed GSTs have very little activity toward AFB1-8,9-exo-epoxide, suggesting that--on a biochemical basis--humans should be quite sensitive to the genotoxic effects of aflatoxins. If a gene encoding a high aflatoxin-active form of GST is present in the human genome, but is not constitutively expressed, and is inducible by dietary antioxidants (as occurs in rats), then chemo- and/or dietary intervention measures aimed at inducing this enzyme could be highly effective. However, as it is possible that human CYP 1A2 may also be inducible by these same chemicals (because of the possible presence of an ARE in this gene), the ultimate consequence of dietary treatment with chemicals that induce biotransformation enzymes via an ARE is uncertain. The balance of the rate of activation (exo-epoxide production) to inactivation (GST conjugation plus other P450-mediated non-epoxide oxidations) may be a strong indicator of individual and species susceptibility to aflatoxin carcinogenesis, if the experimental conditions are reflective of true dietary exposures. There is strong evidence that AFB-8,9-exo-epoxide binds to G:C rich regions of DNA, forming an adduct at the N7-position of guanine. Substantial evidence demonstrates that AFB1-8,9-epoxide can induce activating mutations in the ras oncogene in experimental animals, primarily at codon 12.(ABSTRACT TRUNCATED AT 400 WORDS)

Aflatoxins↗

Potent protection against aflatoxin-induced tumorigenesis through induction of Nrf2-regulated pathways by the triterpenoid 1-[2-cyano-3-,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole.

Synthetic triterpenoid analogues of oleanolic acid are potent inducers of the phase 2 response as well as inhibitors of inflammation. We show that the triterpenoid, 1-[2-cyano-3-,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole (CDDO-Im), is a highly potent chemopreventive agent that inhibits aflatoxin-induced tumorigenesis in rat liver. The chemopreventive potency of CDDO-Im was evaluated by measuring inhibition of formation of putative preneoplastic lesions (glutathione S-transferase P positive foci) in the liver of rats exposed to aflatoxin B1. CDDO-Im produces an 85% reduction in the hepatic focal burden of preneoplastic lesions at 1 micromol/kg body weight and a >99% reduction at 100 micromol/kg body weight. CDDO-Im treatment reduces levels of aflatoxin-DNA adducts by approximately 40% to 90% over the range of 1 to 100 micromol/kg body weight. Additionally, changes in mRNA levels of genes involved in aflatoxin metabolism were measured in rat liver following a single dose of CDDO-Im. GSTA2, GSTA5, AFAR, and EPHX1 transcripts are elevated 6 hours following a 1 micromol/kg body weight dose of CDDO-Im. Microarray analysis using wild-type and Nrf2 knockout mice confirms that many phase 2 and antioxidant genes are induced in an Nrf2-dependent manner in mouse liver following treatment with CDDO-Im. Thus, low-micromole doses of CDDO-Im induce cytoprotective genes, inhibit DNA adduct formation, and dramatically block hepatic tumorigenesis. As a point of reference, oltipraz, an established modulator of aflatoxin metabolism in humans, is 100-fold weaker than CDDO-Im in this rat antitumorigenesis model. The unparalleled potency of CDDO-Im in vivo highlights the chemopreventive promise of targeting Nrf2 pathways with triterpenoids.

Aflatoxin B1↗

Aflatoxins in sera from patients with lung cancer.

Sera from patients with lung cancer and from healthy donors were screened for the presence of aflatoxins. Significant differences in levels of aflatoxins between the two groups were found. Only 1 of the neoplastic patients with aflatoxins in the serum was a smoker. However the percentage of sera from lung cancer containing aflatoxins is not significant enough to provide evidence for a casual relationship between aflatoxins exposure and development of lung cancer in humans.

Aflatoxins↗

Crohn's disease & aflatoxins.

An investigation to examine the relationship between Crohn's disease and aflatoxins, a group of structurally related toxic and carcinogenic metabolites, was carried out on 24 patients. Extracts of serum and urine from the patients were assayed qualitatively by thin layer chromatography and the Aflatest method, and quantitatively by fluorimetry. There was evidence that some patients suffering from Crohn's Disease, together with some having coeliac disease and ulcerative colitis, did have varying amounts of aflatoxins in their serum and urine. The presence of aflatoxins may have been due to exposure to food containing these toxins or inability of the patient to excrete aflatoxins on account of some gastro-intestinal derangement. Only long-term investigation would establish the link between dietary history and the presence of aflatoxins in these patients.

Aflatoxins↗

Morbidity in neonates of mothers who have ingested aflatoxins.

This study was undertaken to assess whether aflatoxin M(1) concentrations in newborn infants correlated with those of their mothers and to determine whether the presence of aflatoxin M(1) in cord blood was associated with an increase in morbidity in the newborn. There was a strong correlation (r =0.797, p <0.0001) between mothers' and cord blood levels of aflatoxin. There was also a strong negative correlation between aflatoxin levels and birthweight (r =-0.565, p <0.001) but there was no association between aflatoxin M(1) concentration in maternal or cord blood and rates of jaundice or infection.

Aflatoxin M1↗

Workgroup report: public health strategies for reducing aflatoxin exposure in developing countries.

Consecutive outbreaks of acute aflatoxicosis in Kenya in 2004 and 2005 caused > 150 deaths. In response, the Centers for Disease Control and Prevention and the World Health Organization convened a workgroup of international experts and health officials in Geneva, Switzerland, in July 2005. After discussions concerning what is known about aflatoxins, the workgroup identified gaps in current knowledge about acute and chronic human health effects of aflatoxins, surveillance and food monitoring, analytic methods, and the efficacy of intervention strategies. The workgroup also identified public health strategies that could be integrated with current agricultural approaches to resolve gaps in current knowledge and ultimately reduce morbidity and mortality associated with the consumption of aflatoxin-contaminated food in the developing world. Four issues that warrant immediate attention were identified: a) quantify the human health impacts and the burden of disease due to aflatoxin exposure; b) compile an inventory, evaluate the efficacy, and disseminate results of ongoing intervention strategies; c) develop and augment the disease surveillance, food monitoring, laboratory, and public health response capacity of affected regions; and d) develop a response protocol that can be used in the event of an outbreak of acute aflatoxicosis. This report expands on the workgroup's discussions concerning aflatoxin in developing countries and summarizes the findings.

Developing Countries↗

Effectiveness of different types of clay for reducing the detrimental effects of aflatoxin-contaminated diets on performance and serum profiles of weanling pigs.

Three trials were conducted with recently weaned pigs (n = 198) to determine the effects of feeding different types of clay in conjunction with aflatoxin-contaminated diets. In Trial 1, pigs (n = 54; trial length 4 wk) were assigned to either an uncontaminated treatment (NC), 800 ppb of aflatoxin from contaminated corn (AC), or AC with one of four clays. In Trial 2 (n = 81; trial length 5 wk), pigs were assigned to NC, AC (500 ppb of aflatoxin from rice starch), or AC with one of seven types of clay. In both trials, pigs fed AC had decreased ADG and gain:feed ratios (P < .05) compared with controls. The clays differed in their ability to produce gains similar to those of controls. The clays did reduce changes in the serum measurements normally affected by aflatoxin, including albumin, total protein, gamma glutamyltransferase (GGT), and alkaline phosphatase (ALP) levels, in a manner similar to their effect on ADG. In Trial 3, pigs (n = 63) were assigned to one of seven diets for 4 wk: NC, AC (800 ppb of aflatoxin) with no clay, AC with one of four levels of a treated Ca bentonite (.25, .5, 1, and 2%), or AC and .5% hydrated sodium calcium aluminosilicate. The addition of treated Ca bentonite to AC improved ADG (P < .05) and ADFI (P < .01) linearly. Gain:feed ratios were not affected by treatments. The inclusion of treated Ca bentonite to the AC diet linearly decreased aspartate aminotransferase (AST) levels and quadratically decreased ALP and GGT levels (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Aflatoxins↗

Potential aflatoxin hazards to human health from direct mold growth on Teleme cheese.

Ninety-four commercial Teleme cheese samples were examined for aflatoxins produced by direct mold growth. The mycoflora on the cheese and in the atmosphere of Teleme cheese plants also were monitored. Penicillium and Aspergillus genera were tested for aflatoxin production after growth on Teleme cheese at 25 degrees C. In all cases, over 78% of the molds were Penicillium species. Aspergillus made up 3.8 to 3.9% and 0 to 7.3% of the mold on cheese and in the plant atmosphere, respectively. None of the commercial samples contained aflatoxins and none of the 448 Penicillium isolates was an aflatoxin producer. Of 22 Aspergillus species, one was capable of producing aflatoxins after direct growth on cheese. Because the physicochemical characteristics of Teleme cheese (high moisture, low pH, and medium salt concentration) favor mold growth, care should be taken to avoid contamination of the cheese by aspergilli.

Aflatoxins↗

The anemia caused by aflatoxin.

The effects of graded doses of dietary aflatoxin (0, 0.625, 1.25, 2.5, 5.0, and 10.0 mug./g.) on hemoglobin, packed blood cell volume, erythrocyte count, leucocyte counts, bone marrow lipid, and bone marrow nucleic acids of chickens were measured. The hemoglobin, packed cell volume, and erythrocyte count were reduced significantly (P less than 0.05) to about the same extent by any given dose. Microscopic examination of stained smears of the bone marrow revealed a hyperplastic response including the granulocytic elements. Chemical analyses of the marrow revealed a decreased lipid content and an increased content of ribonucleic acid and deoxyribonucleic acid. Total leucocytes were increased about three fold by aflatoxin (10 mug./g). Differential leucocyte counts revealed that the heterophils were increased while the eosinophils were unaffected and the basophils, lymphocytes, and monocytes were decreased. The increase in nucleic acids of the marrow and in circulating leucocytes occurred with doses which inhibit growth rate. These data suggest that aflatoxin causes a hemolytic anemia in chickens, that aflatoxin, by itself, is not involved in the hemorrhagic anemia syndrome of chickens, and that aflatoxin does not cause in chickens a general inhibition of ribonucleic acid and protein synthesis as assumed from studies on rats.

Aflatoxins↗

The response of Bobwhite quail chicks to dietary ammonium and an antibiotic-vitamin supplement when fed B1 aflatoxin.

Dietary ammonium phosphate and a water soluble antibiotic-vitamin supplement were tested as possible means of preventing or minimizing the effects of aflatoxin in the diet of Bobwhite quail chicks. Aflatoxin (B1) was added to the diet at levels of 0, 0.4 and .8 ppm in Experiment 1 and 0, .8 and 1.2 ppm in Experiment 2. In addition, ammonium phosphate was included in the diet at levels of 0 or 1.46% and an antibiotic-vitamin supplement was added to the drinking water at 0 or 600 mg/liter. All treatments were given from 1 to 32 days of age in Experiment 1 and 1 to 28 days of age in Experiment 2. The addition of aflatoxin to the diet resulted in decreased body weight, feed consumption and percent tibia ash, increased mortality and increased feed required per unit of body weight. The addition of ammonia to the diet in the form of monoammonium phosphate did not alleviate the effects of aflatoxin. Water supplementation with an antibiotic-vitamin mixture improved weight and feed efficiency and decreased mortality of chicks receiving aflatoxin.

Aflatoxins↗

Individual and combined effects of aflatoxin and deoxynivalenol (DON, vomitoxin) in broiler chickens.

The individual and combined effects of aflatoxin and deoxynivalenol (DON) were evaluated in young broiler chickens (Hubbard X Hubbard). The experimental design was a 2 X 2 factorial with treatments of 0 and 2.5 micrograms of aflatoxin/g of feed (ppm) and 0 and 16 micrograms of DON/g of feed. The broilers were maintained on these dietary treatments from hatching to 3 weeks of age in electrically heated batteries with feed and water available ad libitum. The aflatoxin treatment significantly (P less than .05) decreased body weight; weight gain; increased the relative weight of the spleen, liver, and kidney; induced hepatic hyperlipemia; decreased activity of lactic dehydrogenase; and decreased serum levels of protein, albumin, and phosphorus. The toxicity of DON was expressed through reduced growth rate, increased feed conversion; increased relative weight of the gizzard, anemia, decreased activity of lactic dehydrogenase, and decreased serum triglycerides. The interaction between aflatoxin and DON was characterized by reduced growth rates; increased feed conversion, increased relative weight of the proventriculus, gizzard, spleen, liver, and kidney, anemia, hepatic hyperlipemia, decreased activity of alkaline phosphatase, glutamic oxalacetic transaminase, and lactic dehydrogenase, and decreased serum levels of protein, albumin, uric acid, cholesterol, triglycerides, and calcium. These data demonstrate that both aflatoxin and DON can limit broiler performance and adversely effect broiler health. The effects of the combination of aflatoxin and DON on broiler performance and health was more severe than the individual effects of these mycotoxins; however, the interaction was not severe enough to represent toxic synergy and can best be characterized as additive toxicity.

Aflatoxins↗

Effects of feeding fumonisin B1 present in Fusarium moniliforme culture material and aflatoxin singly and in combination to turkey poults.

Diets containing 200 mg fumonisin B1/kg of feed and .75 mg aflatoxin/kg of feed singly or in combination were fed to female turkey poults (Nicholas Large White) from day of hatch to 21 d of age. When compared with controls, 21-d body weight gains were reduced 10% by fumonisin B1, 39% by aflatoxins, and 47% by the combination. Relative weights (grams/100 g body weight) of the kidney and pancreas increased in poults fed the diet containing aflatoxins alone, whereas the relative weight of the liver decreased. Relative weights of the liver and pancreas increased in the poults fed the fumonisin diet. Relative weights of the kidney, pancreas, and gizzard increased in the poults fed the combination diet, whereas the relative weight of the liver decreased. Most serum constituents, hematology values, and activities of enzymes measured were altered in poults receiving the diets containing aflatoxins with or without fumonisin B1. No major histological lesions were observed in tissues from control poults or poults fed the diet containing fumonisin alone. Lesions associated with aflatoxins were only observed in the liver and occasionally in the kidney of poults fed the diets containing aflatoxins with or without fumonisin B1. The primary hepatic change was bile duct hyperplasia with some hepatocellular degeneration and necrosis and megalocytosis. Occasional necrotic and degenerating tubular epithelial cells were observed in the kidneys. The increased toxicity in poults fed the combination diet for most variables can best be described as additive, although some variables showed less than additive toxicity.(ABSTRACT TRUNCATED AT 250 WORDS)

Aflatoxins↗

Aflatoxin levels in maize and maize products during the 2004 food poisoning outbreak in Eastern Province of Kenya.

BACKGROUND: On 10th May, 2004 food samples suspected to have caused acute poisoning in Makueni district were received at the National Public Health Laboratory Services (NPHLS). On analysis, they were found to be highly contaminated with aflatoxin B1. More cases of poisoning were reported in the district and in neighbouring districts of Kitui, Machakos and Thika. As at 20th July, 2004 the Ministry of Health was aware of 317 cases of which 125 resulted in deaths. OBJECTIVE: To assess the magnitude of aflatoxin contamination of maize and maize products in the affected areas. DESIGN: Random environmental sampling of maize and maize products and case/control samples of the same in the affected regions and subsequent determination of aflatoxin levels using immunoaffinity coupled with solution fluorometry. SETTING: National Public Health Laboratory Services, Ministry of Health, Kenya, from May to August, 2004. SUBJECTS: A total of 480 samples comprising 362 random environmental samples, 26 cases and 92 controls were collected and analysed. The foods analysed included maize grains, maize flour and dehulled dry maize, traditionally known as muthokoi. RESULTS: Forty six point four per cent of the environmental samples, 15% of cases and 29.3% of controls were within the maximum permissible limit of 20 microg implying that over 50% (54.4) of the total did not comply and would be regarded as unfit for human consumption. 6.9% of the environmental samples, 57.7% of cases and 21.7 of controls had levels beyond 1000 microg/Kg. The amount of aflatoxin observed in the food samples had a range of 0-58,000 microg/Kg. CONCLUSION: The population in the affected region was exposed to high levels of aflatoxin. There is need to address the issue of pre and post harvest handling of grain and establishing monitoring and surveillance for early detection and intervention.

Acute Disease↗

Binding of aflatoxin B1 alters the adhesion properties of Lactobacillus rhamnosus strain GG in a Caco-2 model.

Lactic acid bacteria have been previously reported to possess antimycotoxigenic activities both in vitro and in vivo. The objective of this study was to investigate the effect of aflatoxin B1 on adhesion capability of Lactobacillus rhamnosus strain GG using a Caco-2 adhesion model. Removal of aflatoxin B1 by L. rhamnosus strain GG reduced the adhesion capability of this strain from 30% to 5%. It is therefore concluded that aflatoxins may influence the adhesion properties of probiotics able to sequester them, and subsequently these bacteria may reduce the accumulation of aflatoxins in the intestine via increased excretion of an aflatoxin-bacteria complex.

Aflatoxin B1↗

Comparison of Aspergillus ear rot and aflatoxin contamination in grain of high-oil and normal-oil corn hybrids.

High-oil corn (Zea mays L.) grain is a valuable component of feed for monogastric livestock. One method of increasing the concentration of oil in corn grain is the TopCross method. With TopCross, ears of a cytoplasmic male-sterile, normal-oil hybrid are pollinated by a male-fertile, high-oil synthetic hybrid. The concentration of oil in the resulting grain is increased because of xenia effects. Kernels of high-oil corn typically have a larger germ and a smaller endosperm than kernels of comparable normal hybrids. The growth of Aspergillus flavus Link:Fr within germ tissue has been reported to be more extensive than that on the whole corn kernel; therefore, the severity of Aspergillus ear rot could be more extensive and aflatoxin concentrations could be higher in high-oil grain produced by TopCross than in grain with a lower concentration of oil. The objective of this study was to compare Aspergillus ear rot severity levels and aflatoxin concentrations in the grains of hybrids crossed with high-oil or normal-oil pollinators. Fifteen hybrids were evaluated in 1998 and 1999 in Urbana, Ill. Primary ears were inoculated with A. flavus and evaluated for susceptibility to Aspergillus ear rot and aflatoxin production in grain. Concentrations of aflatoxin and oil in corn kernels were significantly higher for high-oil hybrids than for normal-oil hybrids; however, ear rot severity was unaffected by the type of pollinator. These results suggest that grain from high-oil hybrids is at greater risk for aflatoxin contamination during some growing seasons.

Aflatoxins↗

Head space sensor array for the detection of aflatoxin M1 in raw ewe's milk.

A novel screening method was developed for simple and rapid detection of aflatoxin M1 contamination in raw ewe's milk samples without the need for sample pretreatment. The method was based on the use of a commercial head space sensor array system constituted by 12 metal oxide semiconductor sensors, 10 metal oxide semiconductor field-effect transistor sensors, and a pattern recognition software. Twenty-four raw milk samples collected from two different groups of ewes fed with a formulated feed that contained increasing amounts of aflatoxin B1 and six noncontaminated ewe's milk samples were analyzed. The results obtained by using the head space sensor array, processed by statistical methods, made it possible to group the samples according to the presence or the absence of aflatoxin M1. Sample classification was in complete agreement with the aflatoxin M1 content measured by an enzyme-linked immunosorbent assay procedure. This is the first report, to our knowledge, of detection of aflatoxin M1 in ewe's milk by a multisensor array.

Aflatoxin M1↗