Evaluation of chemically characterised essential oils of Coleus aromaticus,Hyptis suaveolens and Ageratum conyzoides against storage fungi and aflatoxin contamination of food commodities

The result of the present investigation explores the efficacy of chemically characterised essential oils (EOs) of Coleus aromaticus, Hyptis suaveolens and Ageratum conyzoides as antifungal and antiaflatoxigenic agent against some storage fungi and the toxigenic strain of Aspergillus flavus (Saktiman 3NSt). Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of these EOs were also determined against the toxigenic strain of A. flavus (Saktiman 3NSt). The EO from C. aromaticus was found to be most effective exhibiting MIC and MFC at 0.1μL mL^?1. The EOs also completely checked aflatoxin B₁ synthesis in concentration‐dependent manner. In addition, fumigation of stored wheat samples with EOs exhibited remarkable protection (>80%) from fungal infestation showing their efficacy during in vivo storage conditions. Based on the results of the present investigation, the EOs of C. aromaticus, H. suaveolens and A. conyzoides may be recommended as novel plant‐based antifungal and aflatoxin B₁ suppressor over the synthetic preservatives.     

Antifungal,antiaflatoxin and antioxidant potential of chemically characterized Boswellia carterii Birdw essential oil and its in vivo practical applicability in preservation of Piper nigrum L. fruits

The study explores the chemical profile, antimicrobial and antioxidant activity of Boswellia carterii essential oil (EO). The EO significantly inhibited growth and aflatoxin production by the food borne toxigenic strain of Aspergillus flavus at 1.75 μl/ml and 1.25 μl/ml respectively. It exhibited broad fungitoxic spectrum against 12 food borne moulds and also showed strong antioxidant activity, IC₅₀ value and % inhibition of linoleic acid peroxidation being 0.64 μl/ml and 51.68% respectively. The antifungal action of EO was observed in terms of reduction in ergosterol content of plasma membrane of A. flavus. As fumigant in food system in storage containers, the EO provided 65.38% protection against fungal deterioration of Piper nigrum. GC–MS results revealed 31 components of EO. The chemically characterized B. carterii EO may thus be recommended as plant based preservative in view of its antifungal, antiaflatoxigenic, antioxidant activity and efficacy in food system.     

Assessment of Thymus vulgaris L. essential oil as a safe botanical preservative against post harvest fungal infestation of food commodities

A total of 14 odoriferous angiospermic essential oils were tested against the toxigenic strain of Aspergillus flavus. The essential oil of Thymus vulgaris L. showed highest antifungal efficacy. The thyme oil absolutely inhibited the mycelial growth of A. flavus at 0.7 μl ml^− 1 and exhibited a broad fungitoxic spectrum against eight different food contaminating fungi viz. Fusarium oxysporum, Cladosporium herbarum, Curvularia lunata, Aspergillus terreus, Aspergillus niger, Aspergillus fumigatus, Alternaria alternata and Botryodiploidia theobromae. The oil also showed significant antiaflatoxigenic efficacy as it completely arrested the aflatoxin B₁ production at 0.6 μl ml^− 1. Thyme oil as fungitoxicant was also found superior over most of the prevalent synthetic fungicides. The LC₅₀ of thyme oil against mice was recorded as 7142.85 μl kg^− 1 body weight indicating its non-mammalian toxicity and strengthening its safe exploitation as preservative for stored food commodities. The findings recommend the thyme oil as potential botanical preservative in eco-friendly control of biodeterioration of food commodities during storage.

Industrial relevance

The thyme essential oil may be recommended for large scale application as a plant based preservative for stored food items because of its strong antifungal as well as antiaflatoxigenic efficacy. Because of broad antimicrobial spectrum, more efficacy over prevalent synthetic preservatives as well as non-mammalian toxicity, the thyme essential oil may be formulated as a safe and economical plant based preservative against post harvest fungal infestation and aflatoxin contamination of food commodities.     

Efficacy of Acorus calamus L. essential oil as a safe plant‐based antioxidant,Aflatoxin B1 suppressor and broad spectrum antimicrobial against food‐infesting fungi

The study explores the efficacy of Acorus calamus L. essential oil (EO) as a safe plant‐based broad spectrum antifungal, antiaflatoxin, antioxidant food additive. The oil completely inhibited the growth and toxin production of the toxigenic strain of Aspergillus flavus at 0.4 and 0.25 μL mL^?1, respectively. EO exhibited pronounced antifungal activity against sixteen food‐infesting fungal species at 0.5 μL mL^?1. The EO showed strong antioxidant efficacy (IC₅₀ 1.06 μL mL^?1) and nonphytotoxic nature on germination of chickpea seeds. The EO was found nonmammalian toxic showing high LD₅₀ (4877.4 μL kg^?1) for mice (oral, acute). The chemical profile of EO was determined through GC and GC–MS analysis. The findings strengthen the possibility of A. calamus EO as a plant‐based food additive in view of its favourable safety profile, antioxidant and antiaflatoxigenic efficacy and broad spectrum antimicrobial activity against food‐infesting fungi.     

Use of essential oil from Mentha arvensis L. to control storage moulds and insects in stored chickpea

BACKGROUND: Fungal contamination and Callosobruchus infestation results in qualitative and quantitative losses of chickpea seeds during storage. Most of the synthetic chemicals used as preservatives have adverse effects. Therefore, the antifungal and insecticidal potential of Mentha arvensis essential oil was evaluated to determine whether this could be an eco‐friendly substitute of synthetic preservatives. RESULTS: The stored chickpea seeds were dominated by Aspergillus flavus (46.1%) and 30% isolates among them were found toxigenic. The MIC of Mentha oil against A. flavus was recorded at 400 µL L^?1 and it exhibited broad fungitoxic activity against 14 storage fungi. The oil was found superior to some prevalent synthetic fungicides. Mentha oil showed potent insecticidal activity against Callosobruchus chinensis at different concentrations and exposure times. The oviposition by C. chinensis was completely checked at 10 µL L^?1 while F₁ emergence was completely inhibited at 200 µL L^?1. During in situ experiments, 94.05% protection of the chickpea from C. chinensis by Mentha oil showed superiority over the organophosphate insecticide malathion, where 90.75% protection was recorded. CONCLUSION: The Mentha EO showing potent fungitoxic and insecticidal efficacy and may be recommended as a plant‐based preservative in the management of fungal and insect infestation of chickpea and other pulses during storage. Copyright © 2009 Society of Chemical Industry     

Efficacy of chemically characterized Ocimum gratissimum L. essential oil as an antioxidant and a safe plant based antimicrobial against fungal and aflatoxin B1 contamination of spices

The paper reports the essential oil (EO) of Ocimum gratissimum as plant based preservative and recommends its application as a nontoxic antimicrobial and antiaflatoxigenic agent against fungal and aflatoxin contamination of spices as well their shelf life enhancer in view of its antioxidant activity. The EO exhibited antifungal activity against fungal isolates from some spices and showed better efficacy as fungitoxicant than prevalent fungicide Wettasul-80. The EO also completely checked the aflatoxin B₁ (AFB₁) synthesis by the toxigenic strains LHP-6 and LHP-10 of A. flavus isolated from Piper nigrum and Myristica fragrans respectively at 0.6 ??l/ml and 0.5 ??l/ml, respectively. In addition, EO showed antioxidant activity through DPPH free radical scavenging and ??-carotene-linoleic acid bleaching assay. Methyl cinnamate (48.29%) and ??-terpinene (26.08%) were recorded the major components of the oil through GC-MS analysis. The EO was found non-mammalian toxic showing high LD₅₀ (11622.67 ??l/kg) during oral toxicity on mice.     

Postharvest incidence of aflatoxins in Linum usitatissimum seeds

Linseed (Linum usitatissimum L), collected from kothi and gunny bag, had a total of 18 fungal species. Aspergillus spp were the more frequent and accounted for more than 40 % of the total count. The gunny bag storage system exhibited higher dominance of A flavus than occurred in the kothi system. Out of 251 isolates screened, 44 % were toxigenic and produced different components of alfatoxins in the range 105–3000 μg litre ^?1. Of 105 samples extracted for the natural presence of alfatoxin, 46 had aflatoxin contamination. The concentration of alfatoxin B₁ in contaminated samples ranged from 120 to 810 μg kg^?1.     

Fungi associated with post-harvest rot of sweet orange (Citrus sinensis) and aflatoxin B1 production by isolates of Aspergillus flavus on plain and supplemented orange juice

Samples of rotting sweet orange (Citrus sinensis) were obtained from the depots, sales counters and waste baskets. Fungi associated with rotting fruits were isolated and identified. Out of 12 species of fungi isolated, 8 are known to be producers of toxins. The 7 isolates of Aspergillus flavus obtained were screened for aflatoxin production in a nutrient solution, and 4 were found to be aflatoxigenic, producing primarily aflatoxin B₁. Aflatoxin B₁ production of the toxigenic isolates were further studied on plain juice and juice separately supplemented with 2.0% yeast extract and 2.0% sucrose. The highest yield of aflatoxin B₁ was produced on juice supplemented with yeast extract by the 4 toxigenic A. flavus isolates, followed by sucrose supplementation while the lowest amount of aflatoxin B₁ was detected on plain juice. Optimum temperature for aflatoxin B₁ production by A. flavus isolate (IBA-O1) was 25 °C to 30 °C, for an incubation period of 7–11 days on plain and supplemented juice media.     

Potential of aflatoxin B1 production by Aspergillus flavus strains on commercially important food grains

In this study, we investigated the potential of aflatoxin B₁ (AFB1) production by five Aspergillus flavus strains previously isolated from sorghum grains on cereals (barley, maize, rice, wheat and sorghum), oilseeds (peanuts and sesame) and pulses (greengram and horsegram). Five strains of A. flavus were inoculated on all food grains and incubated at 25 °C for 7 days; AFB1 was extracted and estimated by enzyme‐linked immunosorbent assay. All A. flavus strains produced AFB1 on all food grains ranging from 245.4 to 15 645.2 μg kg^?1. Of the five strains tested, strain Af 003 produced the highest amount of AFB1 on all commodities ranging from 2245.2 to 15 645.2 μg kg^?1. Comparatively, the AFB1 accumulation was high on rice grains ranging from 3125.2 to 15 645.2 μg kg^?1, followed by peanuts ranging from 2206.2 to 12 466.5 μg kg^?1. Less AFB1 accumulation was observed in greengram and sesame seeds ranging from 645.8 to 2245.2 and 245.4 to 2890.6 μg kg^?1, respectively. Our results showed that all food grains tested are susceptible to A. flavus growth and subsequent AFB1 production.     

Mycoflora and aflatoxin contamination in shelled pistachio nuts

A total of 143 pistachio nut samples collected during harvest, storage and processing were examined for mould growth and aflatoxin production. The mould count was in the range of 10³?10⁴ cfu g^?1 and 10⁵?10⁶ cfu g^?1 for the harvest and storage samples, respectively. The growth of Aspergillus flavus was 38-5-39-5% on the surface of the shells and 6–16% on the kernels without aflatoxin production. The contamination level of A flavus varied among samples collected from different regions. Peeling off the soft shell of pistachio nuts by hand reduced the contamination risk of A flavus to kernels. The predominant flora on stored pistachio nuts were Aspergillus, Penicillium, Cladosporium, Rhizopus, while the genera of Ulocladium, Trichothecium, Aureobasidium and Eurotium were less frequent. Thirty-five percent of the A flavus isolates produced aflatoxins on synthetic media.     

Assessment of chemically characterised Rosmarinus officinalis L. essential oil and its major compounds as plant‐based preservative in food system based on their efficacy against food‐borne moulds and aflatoxin secretion and as antioxidant

The study explores antifungal, anti‐aflatoxigenic and antioxidant efficacy of Rosmarinus officinalis essential oil (ROEO) and its major compounds. In addition, the mode of action of ROEO and its practical efficacy as preservative have been assessed. GC‐MS analysis of ROEO identified 16 compounds; α‐pinene, 1,8‐cineole and camphor being the major compounds. The minimum concentration for inhibition of growth and aflatoxin B₁ secretion against A. flavus (LHP‐6) was found to be 1.5, >5.0, 4.0 and 3.0 μL mL^?1 and 1.25, >5.0, 3.5 and 3.0 μL mL^?1 for ROEO, α‐pinene, 1,8‐cineole and camphor, respectively. The IC₅₀ value through DPPH analysis and percentage inhibition of linoleic acid peroxidation of ROEO were 0.042 μL mL^?1 and 71.05%, respectively. The targeted site of antifungal action of ROEO was confirmed as plasma membrane through ergosterol measurement and TEM analysis. Moreover, ROEO significantly protected Piper nigrum fruits against mould infestation upto 6 months in in vivo trial.     

Antifungal properties and inhibitory effects upon aflatoxin production by Zingiber officinale essential oil in Aspergillus flavus

In this study, the efficacy of ginger (Zingiber officinale Roscoe) essential oil (GEO) in reducing A. flavus growth and aflatoxin production was investigated. Gas chromatography coupled to mass spectrometry and nuclear magnetic resonance spectroscopy showed that the major components of GEO were α‐zingiberene (23.85%) and geranial (14.16%). Mycelial growth of Aspergillus flavus was reduced significantly at a GEO concentration of 150 μg mL^?1, and complete inhibition of conidial germination was observed at a concentration of 10 μg mL^?1. Statistically significant inhibition of ergosterol biosynthesis was detected at a GEO concentration of 10 μg mL^?1. GEO was capable of fully inhibiting aflatoxin production by A. flavus at a concentration of 15 μg mL^?1. The results suggest that low concentrations of GEO are capable of inhibiting aflatoxin production; such ability could be valuable in the upcoming future for agricultural companies to better control aflatoxigenic fungi in agricultural products.     

Selected plant essential oils and their main active components,a promising approach to inhibit aflatoxigenic fungi and aflatoxin production in food

Recent research has showed that Aspergillus flavus and Aspergillus parasiticus are aflatoxigenic species that can become very competitive in the framework of climate change. Aflatoxins show carcinogenic, mutagenic, immunotoxic and teratogenic effects on human and animals. Effective and sustainable measures to inhibit these species and aflatoxins in food are required. Origanum vulgare and Cinnamomum zeylanicum essential oils (EOs) and their major active constituents, carvacrol and cinnamaldehyde, respectively, were assayed for inhibiting these species and aflatoxin production in maize extract medium under different environmental conditions. Doses of 10–1000 mg l^?1 were assayed and the effective doses for 50 (ED₅₀) and 90% (ED₉₀) growth inhibition were determined. The ED₅₀ of cinnamaldehyde, carvacrol, oregano EO, and cinnamon EO against A. flavus were in the ranges 49–52.6, 98–145, 152–505, 295–560 mg l^?1 and against A. parasiticus in the ranges 46–55.5, 101–175, 260–425 and 490–675 mg l^?1, respectively, depending on environmental conditions. In A. flavus treatments ED₉₀ were in the ranges 89.7–90.5, 770–860 and 820–>1000 mg l^?1 for cinnamaldehyde, carvacrol and cinnamon EO, and in A. parasiticus treatments in the ranges 89–91, 855–>1000 and 900–>1000 mg l^?1, respectively. ED₉₀ values for oregano EO against both species were >1000 mg l^?1. Growth rates of both species were higher at 37 than at 25°C and at 0.99 than at 0.96 a_w. Aflatoxin production was higher at 25 than at 37°C. Stimulation of aflatoxin production was observed at low doses except for cinnamaldehyde treatments. The effectiveness of EOs and their main constituents to inhibit fungal growth and aflatoxin production in contact assays was lower than in vapour phase assays using bioactive EVOH-EO films previously reported.     

Evaluation of some essential oils as botanical fungitoxicants for the protection of stored food commodities from fungal infestation

Essential oils from different parts of 18 plants belonging to 11 families were extracted and tested against two toxigenic strains of Aspergillus flavus Link through the poisoned food technique. The oil of Mentha arvensis was found to be effective against both strains of A. flavus and completely stopped the radial mycelial growth of A. flavus at 0.10 mg mL^?1. It was found to be superior over the synthetic fungicides tested and showed a broad fungitoxic spectrum against A. niger, A. fumigatus, Botryodiplodia theobromae, Cladosporium cladosporioides, Fusarium oxysporum, Helminthosporium oryzae, Macrophomina phaseolina and Sclerotium rolfsii at 0.10 mg mL^?1. The oil completely inhibited the aflatoxin B1 production by the toxigenic strain of A. flavus at 0.05 mg mL^?1. Moreover, the Mentha oil also exhibited potent antioxidant activity in 2,2′‐azino‐bis‐3‐ethylbenzothiazoline‐6‐sulphuric acid (ABTS) bioassay. Keeping in mind the side effects of synthetic pesticides and the global interest in botanical pesticides for plant protection due to their biodegradable nature, M. arvensis oil may be used as a botanical fungitoxicant against fungal attack to stored food commodities. The antiaflatoxigenic and antioxidant nature of the oil suggest the possibility of its exploitation for enhancing the shelf life of stored food commodities. Copyright © 2007 Society of Chemical Industry     

The production of aflatoxins in fresh date fruits and under simulated storage conditions

Sixteen varieties of date fruit (Phoenix dactylifera) at three stages of maturation (Kimri, Rutab and Tamr) were examined for the presence of fungi and analysed for aflatoxins B₁, B₂, G₁ and G₂ and sterigmatocystin. Single samples of each variety were used in the study. Samples as received were initially examined for mycoflora and toxin levels and then stored at 98% relative humidity and 30 °C for 14 days to investigate the effects of possible adverse storage conditions on mycoflora and, in particular, aflatoxin formation. All samples showed an absence of aflatoxins and their precusor, sterigmatocystin, after adverse storage for 14 days, although aflatoxin‐producing Aspergillus flavus isolates were identified in 10 varieties at the first stage of maturation (Kimri). High fungal counts were associated with the Rutab stage and low counts with the Tamr stage. The counts of A flavus ranged from 5.00 to 8.16 log₁₀(cfu g^?1) under simulated storage conditions, and three varieties contained significant levels of aflatoxin B₁ or B₂ ranging from 35 to 11 610 µg kg^?1. Sterigmatocystin was not detected in any of the samples as received or under simulated storage conditions. © 2002 Society of Chemical Industry     

Evaluation of Cinnamomum tamala Oil and Its Phenylpropanoid Eugenol for Their Antifungal and Antiaflatoxigenic Activity

During screening of 20 plant extracts against toxigenic strain of Aspergillus flavus (SK 3NSt), the extract of Cinnamomum tamala was found to exhibit absolute fungitoxic activity (100% growth inhibition). Hence, essential oil of C. tamala was extracted and selected for further investigations. The selected oil was subsequently standardised through physico-chemical and fungitoxic properties. Minimum inhibitory concentration (MIC) of the oil for absolute inhibition of growth of the toxigenic strain of A. flavus (SK 3NSt) was found to be 150 μl/l and oil showed fungicidal nature at its respective MIC. The oil had a broad fungitoxic spectrum. It was found to be absolutely inhibitory to almost all the 11 fungi tested when its fungitoxic spectrum was assayed. The antifungal potency of oil was found to be greater in comparison to some prevalent synthetic fungicides. The oils had the potency to withstand high inoculum density. The oil remained active up to 2 years and was thermostable. In addition, the oil showed significant efficacy in arresting aflatoxin B₁ secretion by the toxigenic strain (SK 3NSt) of A. flavus at 750 μl/l. GC-MS analysis of the oil led to the identification of main components of oil viz. eugenol (45.58%), β-pinene (10.03%), β-myrcene (9.73%), β-ocimene (4.51%), β-costol (3.88%) and thujyl alcohol isomer (2.51%). The efficacy of C. tamala oil as aflatoxin B₁ suppressor is being reported for the first time. Eugenol, the major component of oil showed absolute antiaflatoxigenic efficacy even at 250 μl/l. The high LD₅₀ value (16.94 ml/kg body weight) of oil recorded on mice indicates its non-mammalian toxicity and suggests the recommendation of the oil as a novel and safe post-harvest biological preservative of food commodities for their preservation from contaminating fungi.     

Spread of Aspergillus flavus and aflatoxin accumulation in postharvested maize treated with biocontrol products

Maize is a major staple crop and calorie source for many people living in Sub-Saharan Africa. In this region, Aspergillus flavus causes ear rot in maize, contributing to food insecurity due to aflatoxin contamination. The biological control principle of competitive exclusion has been applied in both the United States and Africa to reduce aflatoxin levels in maize grain at harvest by introducing atoxigenic strains that out-compete toxigenic strains. The goal of this study was to determine if the efficacy of preharvest biocontrol treatments carry over into the postharvest drying period, the time between harvest and the point when grain moisture is safe for storage. In Sub-Sahara Africa, this period often is extended by weather and the complexities of postharvest drying practices. Maize grain was collected from fields in Texas and North Carolina that were treated with commercial biocontrol products and untreated control fields. To simulate moisture conditions similar to those experienced by farmers during drying in Sub-Sahara Africa, we adjusted the grain to 20% moisture content and incubated it at 28 °C for 6 days. Although the initial number of kernels infected by fungal species was high in most samples, less than 24% of kernels were infected with Aspergillus flavus and aflatoxin levels were low (<4 ppb). Both toxigenic and atoxigenic strains grew and spread through the grain over the incubation period, and aflatoxin levels increased, even in samples from biocontrol-treated fields. Our molecular analysis suggests that applied biocontrol strains from treated fields may have migrated to untreated fields. These results also indicate that the population of toxigenic A. flavus in the harvested grain will increase and produce aflatoxin during the drying period when moisture is high. Therefore, we conclude that preharvest biocontrol applications will not replace the need for better postharvest practices that reduce the drying time between harvest and storage.     

Assessment of Pelargonium graveolens oil as plant‐based antimicrobial and aflatoxin suppressor in food preservation

BACKGROUND: Contamination of stored food commodities by moulds and mycotoxins results in qualitative as well as quantitative losses. Most of the synthetic antimicrobials used for preservation of stored food items produce side effects in the form of residual and mammalian toxicity. Recently some higher plant products have been recommended as safe alternatives of such synthetic antimicrobials. In the present investigation antifungal efficacy of some essential oils was evaluated against two toxigenic strains of Aspergillus flavus with special reference to the oil of Pelargonium graveolens to investigate its potential to inhibit aflatoxin B₁ secretion. RESULTS: Essential oil of P. graveolens exhibited absolute fungitoxicity against both the toxigenic strains of A. flavus. The minimum inhibitory concentration of the oil was found to be 0.75 g L^?1 and exhibited a fungistatic nature. It was found superior over the synthetic fungicides tested and exhibited a broad fungitoxic spectrum. The oil showed excellent anti‐aflatoxigenic efficacy as it completely inhibited aflatoxin B₁ production even at 0.50 g L^?1. CONCLUSION: This is the first report on the aflatoxin B₁ inhibitory nature of P. graveolens oil. It may be recommended as a novel plant‐based antimicrobial as well as aflatoxin B₁ suppressor over synthetic preservatives in food protection. Copyright © 2008 Society of Chemical Industry     

Efficacy of Some Essential Oils Against Aspergillus flavus with Special Reference to Lippia alba Oil an Inhibitor of Fungal Proliferation and Aflatoxin B1 Production in Green Gram Seeds during Storage

During mycofloral analysis of green gram (Vigna radiata (L.) R. Wilczek) seed samples taken from different grocery stores by agar and standard blotter paper methods, 5 fungal species were identified, of which Aspergillus flavus exhibited higher relative frequency (75.20% to 80.60%) and was found to produce aflatoxin B₁. On screening of 11 plant essential oils against this mycotoxigenic fungi, Lippia alba essential oil was found to be most effective and showed absolute inhibition of mycelia growth at 0.28 μL/mL. The oil of L. alba was fungistatic and fungicidal at 0.14 and 0.28 μL/mL, respectively. Oil had broad range of fungitoxicity at its MIC value and was absolutely inhibited the AFB1 production level at 2.0 μL/mL. Chemical analysis of this oil revealed geranial (36.9%) and neral (29.3%) as major components followed by myrcene (18.6%). Application of a dose of 80 μL/0.25 L air of Lippia oil in the storage system significantly inhibited the fungal proliferation and aflatoxin production without affecting the seed germination rate. By the virtue of fungicidal, antiaflatoxigenic nature and potent efficacy in storage food system, L. alba oil can be commercialized as botanical fungicide for the protection of green gram seeds during storage.     

Evaluation of atoxigenic isolates of Aspergillus flavus as potential biocontrol agents for aflatoxin in maize

Aflatoxin contamination resulting from maize infection by Aspergillus flavus is both an economic and a public health concern. Therefore, strategies for controlling aflatoxin contamination in maize are being investigated. The abilities of eleven naturally occurring atoxigenic isolates in Nigeria to reduce aflatoxin contamination in maize were evaluated in grain competition experiments and in field studies during the 2005 and 2006 growing seasons. Treatments consisted of inoculation of either grains in vials or ears at mid-silking stage in field plots, with the toxigenic isolate (La3228) or atoxigenic isolate alone and co-inoculation of each atoxigenic isolate and La3328. Aflatoxin B₁?+?B₂ concentrations were significantly (p?<?0.05) lower in the co-inoculation treatments compared with the treatment in which the aflatoxin-producing isolate La3228 was inoculated alone. Relative levels of aflatoxin B₁?+?B₂ reduction ranged from 70.1% to 99.9%. Among the atoxigenics, two isolates from Lafia, La3279 and La3303, were most effective at reducing aflatoxin B₁?+?B₂ concentrations in both laboratory and field trials. These two isolates have potential value as agents for the biocontrol of aflatoxin contamination in maize. Because these isolates are endemic to West Africa, they are both more likely than introduced isolates to be well adapted to West African environments and to meet regulatory concerns over their use throughout that region.