A pilot plant for detoxification of aflatoxin B1-contaminated coconut oil by solar irradiation

Summary In coconut oil naturally contaminated with aflatoxin B1, more than 85% of the toxin is present in the soluble form, the remainder occuring in the sediment. This aflatoxin is detoxified when the oil, in a static layer less than 15 mm thick is exposed to solar radiation. A pilot plant, designed to take account of the viscosity and flow characteristics of the oil, was constructed for the exposure of thin layers of oil (2 mm or less) flowing under gravity. At aflatoxin concentrations between 166 and 1250 jug/kg, 75% of the toxin was degraded on exposure to solar radiation of 10 cal/cm²; total detoxification was achieved on repeated exposure. The naturally contaminated coconut oil after exposure to solar radiation did not contain any residual aflatoxins or fluorescent compounds which might have been derived from original aflatoxin B1.

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Resumen Planta pilota para el tratramiento de aceite de coco contaminado con aflatoxina B1 mediante radiation solar y centrifugado En aceite de coco contaminado de forma natural con aflatoxina B1, más del 85% de la toxina se encuentra en forma soluble, el resto permaneciendo en el sedimento. Esta aflatoxina pierde sus propiedades tóxicas cuando se expone el aceite en forma de capa estática de 15 mm de grosor a la radiación solar. Se construyó una planta piloto diseñada teniendo en cuenta la viscosidad y las caracteristicas de fluidez del aceite de forma que pudieran exponerse a la radiación solar capas de aceite muy finas (2 mm o menos) fluyendo gracias a la gravedad. Cuando aceite conteniendo aflatoxina en proporciones entre 166 y 1250 /kg se expuso a una radiación solar de 10 cal./cm² la toxina se degradó en un 75%. La detoxificación total se obtuvo mediante repetición del proceso. El aceite de coco contaminado de forma natural, una vez expuesto a la radiación solar no contenía aflatoxinas residuales ni compuestos fluorescentes potencialmente derivados de la aflatoxina B1 contenida originalmente.

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Résumé Usine-pilote pour la détoxification, par irradiation solaire et centrifugation, de l'huile de coprah contaminée par l'aflatoxine B1 Dans l'huile de coprah spontanément contaminée par l'aflatoxine B1, plus de 85% de la toxine est présente sous forme soluble, le reste se trouvant dans le sédiment. Cette aflatoxine est détoxifiée lorsque l'huile est exposée à la radiation solaire en couche statique de moins de 15 mm d'épaisseur. Une usine-pilote, conçue en tenant compte de la viscosité et de l'écoulement de l'huile, a été construite pour irradier une mince couche d'huile (2 mm ou moins) s'écoulant par gravité. Pour des concentrations en aflatoxine allant de 166 à 1250 g/kg, 75% de la toxine est dégradée par exposition à une irradiation solaire de 10 cal./cm² et, en répétant le traitement, on obtient une détoxification complète. Après exposition à la radiation solaire, l'huile de coprah contaminée spontanément ne contient plus d'aflatoxine résiduelle, ni de composés fluorescents dérivés de l'aflatoxine B1 originelle.

     

Improved method of screening for aflatoxin with a coconut agar medium

Nine isolates of Aspergillus flavus and Aspergillus parasiticus were screened for aflatoxin production on a coconut extract agar medium. Aflatoxin-producing colonies were detected under long-wave UV light (365 nm) by blue fluorescence on the reverse side after 2 to 5 days of growth. Aflatoxin production was verified by chemical analysis. Several types of shredded coconut available in the United States were tested and found to be satisfactory. No additives were required. Various parameters affecting the test were investigated.     

Evaluating the deterioration of peanut seeds contaminated with aflatoxin

Although aflatoxin (AFT) effects on Arachis hypogaea seed germination have been examined, there have been few, if any, investigations correlating germination capacity to field AFT levels for naturally contaminated peanuts. Here, we report this analysis for selected Federal samples containing 0–20 ppb AFT collected at intervals during both September and October 1981. Following the removal of both decoated and split seeds for screening, seeds were germinated for 5 days when both dead and severely infested seeds were discarded. The remaining seeds, normal (uninfested) ‘sprouted’, ‘sprouted’ with some infestation, ‘sprouted’ with morphological abnormalities and normal unsprouted, were both measured and transferred to fresh towels for germination and growth of seedlings. The hypocotyl and primary root lengths of the seedlings were measured. Whereas these lengths ranged from 58·0 ± 6·2 to 68·8 ± 5·6 mm for contaminated seedlings (control 72·5 ± 5·9) at 5 days, hypocotyl and root lengths were 210·0 ± 12·5–225·0 ± 23·0 for contaminated seedlings (control 226·7 ± 21·3) by day 12. Then, percentage germination was 75·0 ± 3·0−84·4 ± 2·5 for toxin seeds control 85·6 ± 1·8). These results suggest that neither the percentage germination nor hypocotyl/ root lengths could be strongly correlated with AFT levels (from natural contamination) for the selected seeds.     

Improved method of screening for aflatoxin with a coconut agar medium.

Nine isolates of Aspergillus flavus and Aspergillus parasiticus were screened for aflatoxin production on a coconut extract agar medium. Aflatoxin-producing colonies were detected under long-wave UV light (365 nm) by blue fluorescence on the reverse side after 2 to 5 days of growth. Aflatoxin production was verified by chemical analysis. Several types of shredded coconut available in the United States were tested and found to be satisfactory. No additives were required. Various parameters affecting the test were investigated.     

Conversion of coconut oil to methyl ketones by two Aspergillus species

Isolates of Aspergillus ruber and A. repens have been grown on coconut oil as the sole carbon source in shake culture. Methyl ketones (C₅-C₁₃) were isolated by solvent extraction and analysed by combined gas chromatography and mass spectrometry. 2-Undecanone was the main volatile product reflecting the high concentration of dodecanoic acid in the original coconut oil. The reactivity of the individual short chain fatty acids as substrates for production of methyl ketones would appear to decrease with increasing molecular weight of the acid after taking into account the greater volatility of the lower molecular weight homologues. 2-Hexanone and 2-octanone were produced by all isolates in low concentration (< 1%). Nonanoic acid and 2-heptanone were converted into 2-octanone. Low concentrations of secondary alcohols were formed under aerobic conditions. It is suggested that the production of methyl ketones by partial β-oxidation is too closely related to mainstream metabolism to be of use in the biochemical taxonomy of the genus.     

Feeding tests with ducklings,Turkey chicks and rabbits and the effects of aflatoxin on these animals

The effects of feeding animals with a toxic strain ofA. flavus was examined. The animals examined were ducklings, turkeys and rabbits. All of them showed loss of weight and mortality after feeding with aflatoxin.The livers of those animals showed that aflatoxin caused pathological changes. This damage of the liver is the lethal effect of the aflatoxin.

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Zusammenfassung Die Wirkung der Fütterung von Tieren mit einem toxischen Stamm vonA. flavus wurde untersucht. Die Tiere sind Entchen, Truthähnchen und Hasen. Alle zeigten einen Gewichtverlust und eine Mortalität nach Aflatoxin-Fütterung. Die Leber der Tiere zeigte, daß Aflatoxin pathologische Veränderung herbeiführte. Die Leberschädigung ist die tötliche Wirkung des Aflatoxins.

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Research supported by grant number FG-Is-161 of the United States Department of Agriculture to whom the author is indebted.     

Effects of aflatoxin and carotenoids on growth performance and immune response in mule ducklings

The purpose of this study was to investigate if carotenoids could alleviate the adverse effects caused by aflatoxin with respect to growth performance and immune response. In two experiments, a total of 320 mule ducklings were assigned to 5 treatments, i.e. control, aflatoxin B(1) (AFB(1)) 200 ppb, AFB(1) +beta-carotene (BC) 200 ppm, AFB(1)+BC 400 ppm, and AFB(1)+astaxanthin (AS) 200 ppm. In experiment 1, the addition of beta-carotene or astaxanthin in the diet containing AFB(1) 200 ppb resulted in a significant decrease in average daily gain as compared with the control. AFB(1) 200 ppb alone and the addition of BC or AS on top of AFB(1) resulted in a significantly lower daily feed intake than for the control group. There were no significant differences in relative organ weights among treatment groups. Both treatments of BC 400 ppm and AS 200 ppm had significantly more macrophages harvested per duck than the control and AFB(1) 200 ppb treatments. However, there were no significant differences among treatments in percentages of phagocytotic macrophages and number of Candida albican phagocytized by phagocytotic macrophages. In experiment 2, blood biochemical parameters and antibody titers were evaluated. There were no significant differences among treatments in total bilirubin content and alkaline phosphatase activity in the serum or in antibody titers against fowl cholera. However, AFB(1) treatment had the highest activities of AST and ALT in the serum. The addition of BC 400 ppm on top of AFB(1) significantly reduced ALT activity as compared with the AFB(1) 200 ppb treatment. These results suggest that carotenoids could provide a slightly toxic alleviating effect on growth performance, enhance the chemotaxis ability of macrophages, and reduce ALT activity elevated by AFB(1).     

A novel strain of Cellulosimicrobium funkei can biologically detoxify aflatoxin B1 in ducklings

Two experiments were conducted to screen microorganisms with aflatoxin B₁ (AFB₁) removal potential from soils and to evaluate their ability in reducing the toxic effects of AFB₁ in ducklings. In experiment 1, we screened 11 isolates that showed the AFB₁ biodegradation ability, and the one exhibited the highest AFB₁ removal ability (97%) was characterized and identified as Cellulosimicrobium funkei (C. funkei). In experiment 2, 80 day‐old Cherry Valley ducklings were divided into four groups with four replicates of five birds each and were used in a 2 by 2 factorial trial design, in which the main factors included administration of AFB₁ versus solvent and C. funkei versus solvent for 2 weeks. The AFB₁ treatment significantly decreased the body weight gain, feed intake and impaired feed conversion ratio. AFB₁ also decreased serum albumin and total protein concentration, while it increased activities of alanine aminotransferase and aspartate aminotransferase and liver damage in the ducklings. Supplementation of C. funkei alleviated the adverse effects of AFB₁ on growth performance, and provided protective effects on the serum biochemical indicators, and decreased hepatic injury in the ducklings. Conclusively, our results suggest that the novel isolated C. funkei strain could be used to mitigate the negative effects of aflatoxicosis in ducklings.     

Enzymatic esterification of starch using recovered coconut oil

Modification of maize and cassava starches was done using recovered coconut oil and microbial lipase. Microwave esterification was advantageous as it gave a DS 1.55 and 1.1 for maize starch and cassava starch, respectively. Solution state esterification of cassava starch for 36 h at 60 °C gave a DS of 0.08 and semi-solid state esterification gave a DS of 0.43. TGA and DSC studies showed that the higher DS attributed to the thermostability, since onset of decomposition is at a higher temperature (492 °C) than the unmodified (330 °C) and was stable above 600 °C. -Amylase digestibility and viscosity reduced for modified starch.     

Cationic amphiphiles with fatty acyl chain asymmetry of coconut oil deliver genes selectively to mouse lung

Recent structure-activity studies have revealed a dramatic influence of hydrophobic chain asymmetry in enhancing gene delivery efficacies of synthetic cationic amphiphiles (Nantz, M. H. et al. Mol. Pharmaceutics2010, 7, 786-794; Koynova, R. et al. Mol. Pharmaceutics2009, 6, 951-958). The present findings demonstrate for the first time that such a transfection enhancing influence of asymmetric hydrocarbon chains observed in pure synthetic cationic amphiphiles also works for cationic amphiphiles designed with natural, asymmetric fatty acyl chains of a food-grade oil. Herein, we demonstrate that cationic amphiphiles designed with the natural fatty acyl chain asymmetry of food-grade coconut oil are less cytotoxic and deliver genes selectively to mouse lung. Despite lauroyl chains being the major fatty acyl chains of coconut oil, both the in vitro and In vivo gene transfer efficiencies of such cationic amphiphiles were found to be remarkably superior (>4-fold) to those of their pure dilauroyl analogue. Mechanistic studies involving the technique of fluorescence resonance energy transfer (FRET) revealed higher biomembrane fusibility of the cationic liposomes of the coconut amphiphiles than that of the symmetric dilauroyl analogue. AFM study revealed pronounced fusogenic nonlamellar structures of the liposomes of coconut amphiphiles. Findings in the FRET and cellular uptake study, taken together, support the notion that the higher cellular uptake resulting from the more fusogenic nature of the liposomes of coconut amphiphiles 1 are likely to play a dominant role in making the coconut amphiphiles transfection competent.     

Earthworm survival in oil contaminated soil

Earthworms are an important component of the soil biota and their response to oil pollution needs to be better understood. Laboratory investigations were undertaken to determine the concentrations of crude oil in soil that leads to death of Lumbricus terrestris and Eisenia fetida and to determine the propensity of L. terrestris to move away from contaminated soil. Clemville sandy clay loam was amended to contain maximum oil contents of 1.5 – 2.5% depending on the particular experiment. Additionally, the ability of L. terrestristo survive in bioremediated oil-contaminated soil was evaluated. An oil content of 0.5% was not harmful to survival of earthworms for 7 d but an oil concentration of 1.5% reduced survival to less than 40%. Bioremediated soil containing 1.2% oil did not reduce survival of L. terrestrisduring 10 d. Survival of L. terrestrisin unweathered oil was improved when free movement between contaminated and uncontaminated soil was possible. Casts of earthworms exposed to oil-containing soil contained 0.2% total petroleum hydrocarbons. An allowable regulatory level of 1% oil contamination in soil may not allow for survival of earthworms.     

Effect of malting and roasting on reduction of aflatoxin in contaminated soybeans

The effect of malting and roasting on the reduction of aflatoxin in contaminated soybeans was studied. Through the observation of the malting process of contaminated and noncontaminated beans it was found that aflatoxin brought about 50% reduction on germination percentage and radicle length of malts. Malting appeared to have a minimal effect in lowering the aflatoxin content of the soybean and aflatoxin G-1 was more affected than B-1 by the treatment. The contaminated samples of nonmalted, steeped and malted soybeans were roasted at 150 ° C and 180 ° C. Roasting at 180 ° C reduced the levels of aflatoxin by 40 to 73 percent depending on the sample nature. Roasting of both steeped and malted beans was much less effective in the reduction of the toxin levels than that of nonmalted beans and in all cases aflatoxin B-1 was more heat resistant than aflatoxin G-1.Present address: North Dakota State University, Cereal Chem. & Techn., Fargo, ND 58105, USA     

不同类型原油污染土壤生物修复技术研究

对不同类型原油污染土壤在实用规模的预制床上采用堆制技术进行生物修复 .通过投加肥料、菌剂、控制水分和pH ,可使微生物获得较好的生态环境 .当稀油、高凝油、特稠油和稠油污染的土壤中原油总量为 2 5 .8～ 77.2 g·kg-1土时 ,经过近 2个月的运行 ,石油总量的去除率可达 38.37%～ 5 6 .74 % .石油中芳烃、沥青和胶质混合物是制约石油快速降解的主要因素 .在处理过程中筛选出石油降解的优势菌株 ,其中有 6株真菌、6株细菌和 1株放线菌 .研究结果为石油污染土壤异位生物修复技术实用化提供了理论依据 .     

Phytotreatment of soil contaminated with used lubricating oil using Hibiscus cannabinus

Soil contamination by hydrocarbons, especially by used lubricating oil, is a growing problem in developing countries, which poses a serious threat to the environment. Phytoremediation of these contaminated soils offers environmental friendly and a cost effective method for their remediation. Hibiscus cannabinus was studied for the remediation of soil contaminated with 2.5 and 1% used lubricating oil and treated with organic wastes [banana skin (BS), brewery spent grain (BSG) and spent mushroom compost (SMC)] for a period of 90 days under natural conditions. Loss of 86.4 and 91.8% used lubricating oil was recorded in soil contaminated with 2.5 and 1% oil and treated with organic wastes respectively at the end of 90 days. However, 52.5 and 58.9% oil loss was recorded in unamended soil contaminated with 2.5 and 1% oil, respectively. The plant did not accumulate hydrocarbon from the soil but shows appreciable accumulation of Fe and Zn in the root and stem of H. cannabinus at the end of the experiment. The first order kinetic rate of uptake of Fe and Zn in H. cannabinus was higher in organic wastes amendment treatments compared to the unamended treatments, which are extremely low. The results of this study suggest that H. cannabinus has a high potential for remediation of hydrocarbon and heavy metal contaminated soil.     

The fate of aflatoxin in naturally contaminated corn during the ethanol fermentation

Corn naturally contaminated with aflatoxin was used as a substrate in the ethanol fermentation. Distribution of toxin in several process and recovery fractions was identified. Although little degradation of the mycotoxin occurred during fermentation, no toxin appeared in the distilled alcohol. As accumulation of toxin in spent grains represents a potential problem in use of the material as animal feed, several decontamination procedures were tested. Sodium hydroxide, ammonium hydroxide, sodium hypochlorite, and hydrogen peroxide were identified as efficient agents of toxin degradation.     

A novel method for non-invasive detection of aflatoxin contaminated dried figs with deep transfer learning approach

Aflatoxins are the most dangerous mycotoxin produced by Aspergillus fungal species, such as Aspergillus flavus and Aspergillus parasiticus, and can cause various health problems, including liver cancer, in humans. Figs and many agricultural products are affected by aflatoxins. It is crucial to detect it before consumption since it is impossible to clean aflatoxin from contaminated foods. Chromatographic methods are considered the gold standard for aflatoxin detection, but these methods are pretty expensive, time-consuming, and destructive. Therefore, various studies have been conducted on non-invasive aflatoxin detection using optical spectroscopic methods. Specifically, aflatoxin-contaminated figs are sorted manually by employees in production facilities using the Bright Greenish Yellow Fluorescence (BGYF) method under ultraviolet (UV) light. However, accurate and safe manual sorting depends on expertise of employees, along with this long exposure time to UV radiation may cause employees skin cancer and eye disorders. This study presents a deep transfer learning-based approach for non-invasive detection and classification of aflatoxin-contaminated dried figs using images captured under UV light. Pre-trained transfer learning models, such as DenseNet, ResNet, VGG, and InceptionNet, are applied to the dataset, but the accuracy of these models does not outperform the other methods that detect aflatoxin with the BGYF method. Therefore, fine-tuning is performed on the models. As a result, training accuracy of 98.57% and validation accuracy of 97.50% is obtained using the DenseNet169 model. The experimental results show that our proposed method achieves the highest accuracy among other methods. Also, the proposed method shows that deep CNN can be used to automatically, rapidly, and effectively detect aflatoxin-contaminated figs.     

Detection of coconut cadang-cadang viroid-like sequences in oil and coconut palm and other monocotyledons in the south-west Pacific

Fractionation, electroblotting and molecular hybridisation of nucleic acids extracted from tissue of African oil palm and coconut palm and some other monocotyledonous species, collected in several areas of the south-west Pacific region, demonstrated the presence of small nucleic acids with nucleotide sequences and secondary structure similar to coconut cadang-cadang viroid (CCCVd). The oil palms which contained CCCVd-related molecules showed orange leaf spots resembling those described for oil palm naturally infected with CCCVd in the Philippines, and also characteristic of a condition known as "genetic orange spotting" (GOS). We provide preliminary evidence that GOS is an infectious disorder caused by a viroid. The coconut palms did not show symptoms typical of cadang-cadang disease, but sometimes were chlorotic, stunted, or had a reduced yield. The possibility that the isolates represent variants of CCCVd is discussed. The data suggest that viroids with nucleotide sequences similar to CCCVd occur widely in palms and other monocotyledons outside the Philippines.