Aflatoxin content and number of fungi in poultry feedstuffs from Indonesia

The content of aflatoxin and associated fungi was determined in 56 samples, including 34 of corn, 10 of soybean meal, nine of rice bran and three of broken rice, collected from different poultry farms and poultry feedmills situated around Jakarta-Bogor, Indonesia.
Ninety-one per cent of the corn samples contained aflatoxins and the total concentration ranged from 22 to 6171 μ g/kg. With rice bran, 100% of the samples were positive for aflatoxin B₁, ranging from 36 to 71 μ g/kg. No aflatoxin was detected in samples of soybean meal or broken rice. All the samples were contaminated by several fungi (8 times 10³–5 times 10⁶cfu/g) and further identification was limited to Aspergillus flavus and A. parasiticus. The dominant species was A. flavus (2 times 10³–4 times 10⁶cfu/g in corn samples, 1·0 times 10³–1·0 times 10⁵cfu/g in soybean meal, 2 times 10⁴–4·4 times 10⁵cfu/g in rice bran and 2 times 10⁴–6 times 10⁴cfu/g in broken rice). Some of the corn samples also contained A. parasiticus (2 times 10³–9·5 times 10⁴cfu/g).     

Validation of an ELISA test kit for the detection of total aflatoxins in grain and grain products by comparison with HPLC

An ELISA Microtiter Plate, Total Aflatoxin Test called AgraQuant^® was validated to measure total aflatoxins in a range from 4 to 40 ppb in corn, corn meal, corn gluten feed, corn gluten meal, corn germ meal, corn/soy blend, popcorn, sorghum, wheat, milled rice, soybeans, peanuts and cottonseed. The test is performed as a solid phase direct competitive ELISA using a horseradish peroxidase conjugate as the competing, measurable entity. For the test method, aflatoxins are extracted from ground samples with 70 % methanol and sample extracts plus conjugate are mixed and then added to the antibody-coated microwells. After 15 min incubation at room temperature, the plate is washed and enzyme substrate is added and allowed to incubate for an additional 5 min. Stop solution is then added and the intensity of the resulting yellow color is measured optically with a microplate reader at 450 nm. Results obtained from internal validation studies assessing accelerated stability indicate a minimum of 1 year shelf life for the kits; accuracy and precision are comparable to HPLC in the range of 0–320 ppb and limit of detection in corn is 2.5 ppb. Comparison of the method to HPLC, ability to detect individual aflatoxins and ruggedness of the test kits at 18–30°C determined this test to be rugged, sensitive, accurate, precise and effective comparable to HPLC for measuring total aflatoxins ranging from 4 to 40 ppb in the commodities evaluated.     

Digestibility of phosphorous in cereals and co-products for animal feed

Feed ingredients used in swine diets contain various levels and availabilities of nutrients. Nutritional precision evaluation of each ingredient is necessary for formulating diets of pigs. Especially, phosphorous (P) is one of important nutrients for metabolism. However, current data of P digestibility were most apparent digestibility. Therefore, this study was aimed to estimate the coefficient of total tract standardized digestibility (CTTSD) of P in cereals and various co-products used in pig diet. Twelve barrows (initial BW?±?SD, 46.70?±?3.21?kg) were used in this experiment. The experimental design was a 12?×?8 incomplete Latin square with 12 diets and 8 periods. Experimental diets were consisted of barley, wheat, lupine kernel (LK), soybean meal (SBM), almond meal (AM), corn gluten meal (CGM), corn gluten feed from China (CGF-C), corn gluten feed from Korea (CGF-C), wheat bran (WB), rice bran (RB), lupine hull (LH) and P-free diet. The CTTAD of Ca was higher in AM than RB and CGF-K. The LK and CGM showed greater CTTSD of P than RB and LH. In conclusion, our results indicated that the cereals and co-products as P sources were the ideally used as an ingredient in mixed diets of the growing-finishing pigs.     

Effect of phytate on aflatoxin formation by Aspergillus parasiticus grown on different grains

Aflatoxin production by Aspergillus parasiticus on corn, soybean, and cottonseed in the absence or presence of added sodium phytate was examined. No variation in aflatoxin concentrations was found in raw, chemically sterilized, or autoclaved soybeans whereas a five-fold reduction in total aflatoxins was found in cottonseed after addition of 330 g sodium phytate to 10 g of autoclaved material. However, phytate did not affect aflatoxin production on non-sterile cottonseeds, although in corn a slight inhibition was found. Extraction of raw soybeans with hexane allowed production of 20-fold more aflatoxins, but levels were still lower than those found on rice or corn. Part of this relative inhibition in soybeans may arise from a heat-unstable, polar solvent-soluble, dialyzable factor present in soybeans. Our results support the conclusion that phytate is not the factor in soy responsible for its relative resistance to aflatoxin formation.     

Germination and post-germination development of Colletotrichum dematium f. circinans on Allium cepa

p6e production of aflatoxins by Aspergillus flavus SRRC-1000 growing on soybean (Forrest) and glandless cottonseed (Deltapine 16) meals was examined with respect to effects of zinc and phytate. Aflatoxins were not produced on unautoclaved soybean meal. Addition of zinc (as zinc sulfate) to autoclaved meal inhibited aflatoxin production and supplementation with sodium phytate relieved this inhibition. Addition of sodium phytate alone promoted production. When cottonseed meal was treated to release native phytate into the meal from phytate-sequestering globoids, aflatoxin production increased. However, the largest production on cottonseed meal occurred upon dialysis of the meal without releasing phytate, implying removal of a small molecular weight inhibitor.     

Survey of Vietnamese Peanuts,Corn and Soil for the Presence of <Emphasis Type="Italic">Aspergillus flavus</Emphasis> and <Emphasis Type="Italic">Aspergillus parasiticus</Emphasis>

Aspergillus flavus and Aspergillus parasiticus cause perennial infection of agriculturally important crops in tropical and subtropical areas. Invasion of crops by these fungi may result in contamination of food and feed by potent carcinogenic aflatoxins. Consumption of aflatoxin contaminated foods is a recognised risk factor for human hepatocellular carcinoma (HCC) and may contribute to the high incidence of HCC in Southeast Asia. This study conducted a survey of Vietnamese crops (peanuts and corn) and soil for the presence of aflatoxigenic fungi and used microsatellite markers to investigate the genetic diversity of Vietnamese Aspergillus strains. From a total of 85 samples comprising peanut (25), corn (45) and soil (15), 106 strains were isolated. Identification of strains by colony morphology and aflatoxin production found all Vietnamese strains to be A. flavus with no A. parasiticus isolated. A. flavus was present in 36.0% of peanut samples, 31.1% of corn samples, 27.3% of farmed soil samples and was not found in virgin soil samples. Twenty-five per cent of the strains produced aflatoxins. Microsatellite analysis revealed a high level of genetic diversity in the Vietnamese A. flavus population. Clustering, based on microsatellite genotype, was unrelated to aflatoxin production, geographic origin or substrate origin.     

Mycoflora,and mycotoxins production in Nigerian corn and corn-based snacks

Thirty-one fungal species, mostly toxigenic and belonging to 11 genera were isolated from corn, corn cake and corn roll snack samples.Aspergillus, Penicillum andFusarium accounted for 10, 6 and 3 of the species and altogether, they constituted 90, 94 and 88 percent of the total fungi in corn, corn cake and corn roll snack respectively. Mycotoxins (aflatoxins and ochratoxin A) were detected in 45, 80 and 12 percent while the means and ranges of the total aflatoxins recorded were: 200(25–770 ppb); 233(15–1070 ppb) and 55(10–160 ppb) for corn, corn cake and corn roll snack samples respectively. Ochratoxin A was detected at toxicologically significant levels in only 15 percent of the corn cake samples analyzed. All the strains ofAspergillus flavus andA. ochraceus tested produced aflatoxin B and ochratoxin A, respectively, when they were cultured on each of the three substrates. In each case, substantial quantities of the toxins were produced from 25 to 35°C with the peak level recorded at 30°C. Toxin production was detected only in substrates with 15 percent moisture content and above; reaching the maximum at 25 or 30 percent moisture level. No substantial differences in the amount of toxins were elaborated with further increase in substrates' moisture content. Of the three substrates, corn cake was the most suitable for aflatoxin B production while they were all equally suitable for the elaboration of ochratoxin A.     

Survey of fungal counts and natural occurrence of aflatoxins in Malaysian starch-based foods

In a survey of starch-based foods sampled from retail outlets in Malaysia, fungal colonies were mostly detected in wheat flour (100%), followed by rice flour (74%), glutinous rice grains (72%), ordinary rice grains (60%), glutinous rice flour (48%) and corn flour (26%). All positive samples of ordinary rice and glutinous rice grains had total fungal counts below 103 cfu/g sample, while among the positive rice flour, glutinous rice flour and corn flour samples, the highest total fungal count was more than 103 but less than 104 cfu/g sample respectively. However, in wheat flour samples total fungal count ranged from 102 cfu/g sample to slightly more than 104 cfu/g sample. Aflatoxigenic colonies were mostly detected in wheat flour (20%), followed by ordinary rice grains (4%), glutinous rice grains (4%) and glutinous rice flour (2%). No aflatoxigenic colonies were isolated from rice flour and corn flour samples. Screening of aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2 using reversed-phase HPLC were carried out on 84 samples of ordinary rice grains and 83 samples of wheat flour. Two point four percent (2.4%) of ordinary rice grains were positive for aflatoxin G1 and 3.6% were positive for aflatoxin G2. All the positive samples were collected from private homes at concentrations ranging from 3.69–77.50 μg/kg. One point two percent (1.2%) of wheat flour samples were positive for aflatoxin B1 at a concentration of 25.62 μ};g/kg, 4.8% were positive for aflatoxin B2 at concentrations ranging from 11.25–252.50 μg/kg, 3.6% were positive for aflatoxin G1 at concentrations ranging from 25.00–289.38 μg/kg and 13.25% were positive for aflatoxin G2 at concentrations ranging from 16.25–436.25 μg/kg. Similarly, positive wheat flour samples were mostly collected from private homes. This revised version was published online in June 2006 with corrections to the Cover Date.     

Improved methodology for detecting aflatoxin production quantitatively in natural media

This report describes a simple, rapid, and quantitative method for screening the aflatoxin production by moulds of theAspergillus flavus group, using a natural media (moist wheat or rice), and a single chloroform extraction for aflatoxins. The aflatoxins were detected and quantified by thin layer chromatography. The methodology proposed was useful in detecting aflatoxin production on the 3rd day of incubation in more than 85 % of the strains studied (both culture collection strains and field isolates).     

Aflatoxin Production of Species and Strains of the Aspergillus flavus Group Isolated from Field Crops

Peanuts, cottonseed, rice, and sorghum from Texas were sampled over a 3-year period. To insure adequate isolation of alfatoxin-producing species of fungi, low-quality lots were sampled at a rate greater than their respective proportional representation. Aflatoxins were found each year in peanut and cottonseed and were found in 2 of 3 years in rice and sorghum. Aflatoxins were detected in all four crops. The Aspergillus flavus group was much more prevalent in peanut and rice than in cottonseed and sorghum. Of the isolates of the A. flavus group, 96% from peanuts, 79% from cottonseed, 49% from sorghum, and 35% from rice produced aflatoxins. The average toxin production of isolates from rice was much less than that from peanuts, cottonseed, or sorghum. More than 90% of all isolates of the A. flavus group were identified as the species A. flavus. A. parasiticus was isolated from all four crops. Only A. parasiticus produced aflatoxin G.     

L-glutaminase production by <Emphasis Type="Italic">Trichoderma koningii</Emphasis> under solid-state fermentation

Solid state fermentation was conducted for the production of L-glutaminase by Trichoderma koningii Oud.aggr. using different agro-industrial byproducts inlcuding wheat bran, groundnut residues, rice hulls, soya bean meal, corn steep, sesamum oil cake, cotton seed residues and lentil industrial residues as solid substrates. Wheat bran was the best substrate for induction of L-glutaminase (12.1 U/mg protein) by T. koningii. The maximum productivity (23.2 U/mg protein) and yield (45.0 U/gds) of L-glutaminase by T. koningii occurred using wheat bran of 70% initial moisture content, initial pH 7.0, supplemented with D-glucose (1.0%) and L-glutamine (2.0% w/v), inoculated with 3 ml of 6 day old fungal culture and incubated at 30°C for 7 days. After optimization, the productivity of L-glutaminase by the solid cultures of T. koningii was increased by 2.2 fold regarding to the submerged culture.     

Apparent digestion coefficients for dry matter, protein and essential amino acids in terrestrial ingredients for Pacific shrimp Litopenaeus vannamei (Decapoda: Penaeidae)

Protein quality mainly depends on the essential amino acid (EAA) profile, but also on its bioavailability, because EAA digestibility is generally lower than the analyzed amounts. This information is needed in the aquaculture industry for aquafeed formulation. For this purpose, the apparent digestibility coefficients of dry matter, protein, and essential amino acids of eight feedstuffs of terrestrial origin were determined for the juvenile whiteleg shrimp Litopenaeus vannamei (15-19 g), using 1% chromic oxide as an inert marker. A reference diet was formulated and produced in the laboratory. Eight experimental diets were prepared each with 30% of one of the experimental ingredients added to the reference diet: casein, porcine byproduct meal poultry byproduct meal, corn meal, wheat gluten meal, soybean paste, sorghum meal, and wheat meal. The experiment consisted of a single-factor, completely randomized design with three replicates per treatment. Samples of ingredients, diets and feces were analyzed for nitrogen and amino acids. For amino acid assay, we used reverse-phase high performance liquid chromatography. To avoid partial loss of methionine and cystine, samples of ingredients, diets, and feces were oxidized with performic acid to methionine sulfone and cysteic acid prior to acid hydrolysis. The apparent dry matter and protein digestive utilization coefficients varied from 68% to 109% and from 70% to 103%, respectively. Apparent digestibility of protein for casein, soy paste, wheat meal and wheat gluten were very high (over 90%), corn gluten and poultry byproducts meal showed high protein digestibility (over 80%), but porcine byproducts meal and sorghum meal had low digestibility (76% and 70%, respectively). There was a reasonable, but not total, correspondence between apparent protein digestibility and average essential amino acid digestibility coefficients, except for arginine in corn gluten, phenylalanine and leucine in sorghum meal, phenylalanine in soy paste and lysine in wheat meal and poultry by-product meal. The most digestible feed ingredients for whiteleg shrimp were: wheat gluten, wheat meal and soy paste; poultry byproduct meal and corn gluten were less digestible and the lowest digestibility occurred in porcine byproduct meal and sorghum meal. Feedstuffs exhibited great variability in dry matter, protein and amino acid digestive utilization coefficients, which should be considered when formulating shrimp feeds.     

Comparative study on concentrations of deoxynivalenol and zearalenone in kernels of transgenic Bt maize hybrids and nontransgenic maize hybrids

A survey of aflatoxin contamination in selected Colombian foods was conducted over a 12-month period on a total of 248 samples. Samples were collected in supermarkets, retail stores and stock centres and were grouped into five categories: (1) corn and corn products, (2) cereal grains, (3) rice and rice products, (4) legume seeds; and (5) snacks and breakfast cereals. Aflatoxins were identified and quantitated using a liquid chromatographic technique with a limit of detection of 1 ng/g for each aflatoxin. Aflatoxins were detected in 14 of 109 samples of corn and corn products, 4 of 40 samples of rice and rice products, 2 of 30 samples of legume seeds, and 2 of 11 samples of snacks and breakfast cereals. None of the cereal grains samples analysed contained detectable levels of aflatoxins. Twelve of the total of 22 positive samples exceeded the maximum tolerable level of aflatoxin B₁ adopted in most countries (5 ng/g); 10 of these 12 samples corresponded to corn and corn products. The results of the present study indicate that aflatoxin B₁ contamination in certain foods in Colombia is a major public health concern. Continuous monitoring of aflatoxin B₁ levels in Colombian foods is advised.     

青霉PT95菌固态发酵产生类胡萝卜素的研究

本文对青霉Ｐｅｎｉｃｉｌｌｉｕｍｓｐ．ＰＴ９５菌株在固态发酵条件下菌核内产生类胡萝卜素进行了初步研究。结果表明,在３种固态发酵培养基中,玉米粉培养基（ＳＭＡ）比麸皮２基和棉籽壳培养基更适合于ＰＴ９５菌株固态发酵产生类胡萝卜素。为了增加菌核干重和提高类胡萝卜素产率,ＳＭＡ中需要添加氮源、碳源和植物油。在所度的各种氮、碳源中,以硝酸钠和麦芽糖效果最佳。通过我试验确定了在培养基盐溶液中添加硝酸钠３ｇ／Ｌ     

Bright Greenish-Yellow Fluorescence and Aflatoxin in Agricultural Commodities

The corn milling industry has widely accepted the presence of bright greenish-yellow fluorescence under a black light as a presumptive indicator of aflatoxin (a poison produced by the mold Aspergillus flavus). This test was applied to wheat, oats, barley, rice, coconut, white corn, yellow corn, peanuts, sorghum, and soybeans, and evaluated in the laboratory. Our study supported the use of bright greenish-yellow fluorescence as a presumptive test for aflatoxin in wheat, oats, barley, corn, and sorghum.     

The effect of dietary aflatoxin on wild turkey poults

Aflatoxins, toxic metabolites of Aspergillus flavus or Aspergillus parasiticus, cause poor feed utilization, decreased weight gains, depressed immune function, liver dysfunction, coagulation abnormalities, and death in a wide variety of species including humans. Conservationists have become concerned that increasingly popular wildlife feeding or baiting practices could expose wildlife to toxic amounts of aflatoxin-contaminated grains. In particular, the effects of aflatoxins on the wild turkey (Meleagris gallopova silvestris) are of concern because the conspecific domestic turkey is highly susceptible to aflatoxins. To evaluate the effect of dietary aflatoxin on wild turkeys, four groups of 4-mo-old wild turkeys were fed diets containing either 0, 100, 200, or 400 micrograms aflatoxin/kg feed for 2 wk in September and October 1996. Aflatoxin-fed poults had decreased feed consumption and weight gains as compared with control poults. Decreased liver-to-body weight ratios, liver enzyme alterations, slightly altered blood coagulation patterns, and mild histologic changes indicated low-level liver damage. Compromise of cell-mediated immunity was indicated by decreased lymphoblast transformation. The effects were apparent in all treatment groups to variable levels, but significant differences most often were found at 400 micrograms aflatoxin/kg feed. This study shows that short-term aflatoxin ingestion by wild turkeys can induce undesirable physiologic changes; therefore, exposure of wild turkeys to feeds containing aflatoxin levels of 100 micrograms aflatoxin/kg feed or more should be avoided.     

Effects of Moisture Content and Temperature on Aflatoxin Production in Corn

Samples of freshly harvested and remoistened corn, of various moisture contents, were stored at different temperatures; analyses for aflatoxin content were made periodically. At moisture levels above 17.5% and at temperatures of 24 C or warmer, aflatoxins were formed by Aspergillus flavus present in the original epiphytic mycoflora. Remoistened dried corn was subject to more rapid fungal deterioration and aflatoxin formation than freshly harvested corn. Screening of the fungi present in the corn revealed aflatoxin production only by A. flavus. The toxigenic strains produced only aflatoxins B(1) and B(2).     

Formulating Atoxigenic Aspergillus flavus for Field Release

A procedure was developed to encapsulate mycelia of an atoxigenic strain of Aspergillus flavus in alginate pellets for seeding into agricultural fields in order to reduce aflatoxin contamination via competitive exclusion. Kaolin, a clay filler commonly employed in alginate formulations, was detrimental to pellet performance as measured by spore yield. Corn cob grits, a by-product of the corn industry, was found to be an excellent replacement for kaolin. Of nine nutritive adjuvants tested, wheat gluten improved pellet performance the most, although gluten concentrations above 5% were difficult to process. The best formulation tested consisted of 1% sodium alginate, 5% corn cob grits and 5% wheat gluten. On a 'per gram' basis, this alginate formulation yielded more spores than either A. flavus sclerotia or colonized wheat seed. Pesticides were also tested as adjuvants with potential use for protecting pellets under field conditions. Only one (chloramphenicol) of four tested pesticides (the others were dichloran, rose Bengal and cyfluthrin) reduced pellet sporulation. Formulations with or without pesticide adjuvants retained similar spore yield potential during a 2-year storage at 8 C. However, spore production in stored products lagged behind that of fresh products. At 75% relative humidity (RH), pellet storage stability decreased with increasing temperature from 27 to 42 C. Pellet spore yield at 32 C decreased as RH decreased from 100 to 90%. Sporulation occurred at 90% RH but not at 88% RH. Spore yield varied widely in four field tests, and the cumulative spore yield was inversely correlated (r2= -0.798, P 0.01) with rainfall. The results suggest that alginate pellets may be effective formulations for delivery of atoxigenic A. flavus strains to furrow-irrigated cotton in desert environments, where aflatoxin contamination of cottonseed is most severe.     

Mycobiota and mycotoxins in fermented feed,wheat grains and corn grains in Southeastern Buenos Aires Province,Argentina

Wheat (as bran) and corn (as dry grain or fermented feed) are main ingredients of feedstuffs used in local cattle and pig farms in the South of the Buenos Aires Province (Argentina). Therefore, determining mycobiota and mycotoxins in wheat and corn is of prime importance for developing feed management techniques to optimise animal production and to minimize toxicity. Then, a mycological survey was carried out in the Southeastern part of the Buenos Aires Province, in order to identify the mycobiota and the main mycotoxins present in fermented feed, wheat grain and corn grain samples. Samples were cultured for fungal quantification, isolation and identification, and analysed for deoxynivalenol (DON), zearalenone (ZEA), T-2 toxin and aflatoxins (AFLA). Penicillium (74%), Aspergillus (32%) and Scopulariopsis (21%) were the prevalent genera in fermented feed. Penicillium (70%), Fusarium (47%) and Aspergillus (34%) were the most frequent fungi isolated from corn. Penicillium (42%), Fusarium (27%) and Alternaria (25%) were the most frequently recovered genera from wheat. DON was detected in 59% of the corn samples, in 45% of the wheat samples and in 38% of the silage samples. ZEA was detected in 36% of the corn samples, in 49% of the wheat samples and in 16% of the silage samples. T-2 toxin and aflatoxin B1 were each detected in 4% of the corn samples. Eighteen percent of the fermented feed samples showed T-2 contamination. Fermented feed and wheat samples were negative for AFLA.     

Effect of culture substrates on virulence of Beauveria bassiana (Ascomycota: Cordycipitaceae) conidia against the browntail moth,Euproctis chrysorrhoea (Lepidoptera: Lymantriidae)

We studied the effect of different solid substrates on virulence of two Beauveria bassiana isolates against the browntail moth, Euproctis chrysorrhoea (L.) (Lep.: Lymantriidae). Conidia produced on wheat grains, wheat flour, wheat bran, rice flour, rice bran, rice paddy, corn flour, millet, and Sabouraud's dextrose agar with 1% yeast extract (SDAY) as control were compared. There were significant differences among these substrates for their effects on the virulence of produced conidia. Applying 10⁷ conidia/mL of B. bassiana EUT105, produced on rice bran caused the highest (84.9%) and on rice flour, the lowest (57.6%) mortalities. Bioassay on fifth-instar larvae using aerial conidia harvested from wheat grains, rice paddy, and SDAY indicated that conidia from wheat grains had the highest virulence while those from rice paddy, the lowest.