Antifungal, aflatoxin inhibition and antioxidant activity of Callistemon lanceolatus (Sm.) Sweet essential oil and its major component 1,8-cineole against fungal isolates from chickpea seeds

The present investigation reports the extent of molds and aflatoxin contamination to Avarodhi, Kabuli, Pusa 256, Radha and Samrat varieties of chickpea seeds. The study also examines the chemical composition of Callistemon lanceolatus (Sm.) Sweet essential oil and its antifungal, antiaflatoxin and antioxidant activity. During standardization of chemical profile, a total of 8 compounds constituting 0.862 mg/mL of oil composition were analyzed by GC-MS analysis where 1,8-cineole was recorded as a major component (0.56 mg/mL). The antifungal activity of EO and 1,8-cineole was evaluated by contact assay on Czapek’s dox agar. The EO (0.227-0.908 mg/mL) and 1,8-cineole (0.918 mg/mL) showed remarkable antifungal effect against all the fungal isolates of chickpea. Their minimal inhibitory (MIC) and fungicidal (MFC) concentrations for Aspergillus flavus were lower than those of the prevalent systemic fungicide Nystatin. Aflatoxin B₁ (AFB₁) production by NKD-208 isolates of A. flavus was strongly inhibited even at the lower fungistatic concentration of EO and 1,8-cineole.There was no adverse effect of EO treatment on chickpea seed germination suggesting its non-phytotoxic nature. Based on the findings of present investigation, C. lanceolatus essential oil may be recommended as botanical preservative for the enhancement of shelf life of food items in- view of the adverse effect of synthetic preservatives and its strong antifungal, aflatoxin inhibition and antioxidant activity.     

Biological activities of Boswellia sacra extracts on the growth and aflatoxins secretion of two aflatoxigenic species of Aspergillus species

Aflatoxins are the most serious carcinogenic, hepatotoxic, teratogenic and mutagenic secondary metabolites which adversely affect human and animal health. This study was designed to evaluate the in vitro inhibitory effect of different concentrations of Boswellia sacra resin (2.5, 5, 7.5 and 10 g/100 ml), leaf extract (5, 7.5, 10, 12.5 and 15 ml/100 ml), and essential oil (1, 2, 3, and 4 ml/100 ml) on the growth and aflatoxins production by two species of Aspergilli, namely Aspergillus flavus (SQU21) and Aspergillus parasiticus (CBS921.7). Resin of B. sacra caused 57.9–92.1% inhibition of aflatoxin secretion by A. flavus and 43.6–95.7% for A. parasiticus. However, the mycelial dry weights were significantly increased by 20.9–52.7% for A. flavus, and 8.9–68.5% for A. parasiticus. The leaf extract of B. sacra apparently enhanced aflatoxins production by 20–50%, and mycelial dry weight by 25.5–29.1% for A. flavus and A. parasiticus. The essential oil of B. sacra at different concentrations similarly inhibited the fungal growth and aflatoxins production by 45.8–83.7% for A. flavus and 41.3–83.5% for A. parasiticus which indicates the antifungal activity of this oil. None of the B. sacra extracts detoxified pure aqueous aflatoxin B₁. We have concluded that B. sacra resin and essential oil possess biological activity against biochemical synthesis and metabolic pathway of aflatoxin production of the two Aspergillus species. Therefore, the resin and essential oil of B. sacra can be recommended as safe plant based bioreservatives to enhance shelf life of food and feed products with reference to adverse effect of physical and synthetic chemical preservatives and their antimicrobial and aflatoxins inhibition activity.     

Aflatoxigenic fungi and aflatoxins occurrence in sultanas and dried figs commercialized in Brazil

The presence of aflatoxins, Aspergillus flavus and Aspergillus parasiticus in dried fruits was investigated. A total of 62 dried fruit samples were analyzed (24 black sultanas, 19 white sultanas and 19 dried figs). A total of 10 A. flavus isolates were found, nine in one white sultana sample (corresponding to 18% infection) and one isolate in dried figs (2%), and all of them were aflatoxin B₁ and B₂ producers. A. parasiticus was not found. Aflatoxins were detected in 3 of 19 (16%) white sultana samples analyzed and, the limits were not higher than 2.0 μg/kg. In dried figs 11 of 19 (58%) samples were contaminated with aflatoxins and, with exception of one sample that was contaminated with 1500 μg/kg of B₁ aflatoxin, the others had less than 2.0 μg/kg. Neither aflatoxigenic or aflatoxins contaminated black sultanas.     

Mycoflora and absence of aflatoxin contamination of commercialized cassava chips in Benin, West Africa

Studies conducted in Benin, in which the main staple foods are maize, cassava, groundnuts and yams, showed high levels of aflatoxin residues in blood of the exposed population. The natural contamination with fungi and aflatoxins in cassava chips sold at markets in Benin, West Africa was investigated. A total of sixty samples were sampled from open markets in 11 districts of 3 agroecological zones and analyzed for the presence of mycoflora and aflatoxin B₁, B₂, G₁ and G₂. Fourteen genera of fungi were associated with marketed dried cassava chips. Within these, twenty- two isolates were identified to species level, whereas four were identified only to genus. The dominating fungal species isolated were Rhizopus oryzae, Nigrospora oryzae, Chrysonilia sitophila, Cladosporium resinae, Cladosporium herbarum, Apergillus niger and Aspergillus flavus. Fifty-four out of sixty samples were contaminated with A. flavus. The rate of occurrence in CFU/g of A. flavus fungi was lower than for all other fungal species together. Aflatoxin was not detected in any of the samples analyzed using HPLC with post-column photochemical derivatization and fluorescence detection. The limit of detection (LOD) was 0.1 μg/kg. Results from this study suggest cassava chips are unlikely to be a source of aflatoxin in Benin, and that other staples such as maize and groundnuts are more important in aflatoxin exposure. Therefore it can be speculated that staples like maize and groundnut are more important in aflatoxin exposure.     

Incidence and consumer awareness of toxigenic Aspergillus section Flavi and aflatoxin B1 in peanut cake from Nigeria

Peanut cake samples were collected from major markets in five states of Nigeria and evaluated for incidence of toxigenic Aspergillus section Flavi populations, and aflatoxin B₁ (AFB₁) levels by liquid chromatography tandem mass spectrometry (LC–MS/MS). The awareness of consumers to the presence of aflatoxin in the snack and potential health risks of its regular ingestion was evaluated by questionnaire analysis. Aspergillus section Flavi populations were recovered from 83% of the peanut cake samples. Aspergillus flavus L-strain was the most predominant (>56%) across the states while Aspergillus tamarii had the least mean incidence (2.7%). The incidence of atoxigenic strains was significantly (p < 0.05) higher than that of toxigenic strains in samples from Lagos and Kaduna, while the toxigenic strains had a significantly (p < 0.05) higher incidence than the atoxigenic strains in Niger. All analyzed cake samples contained AFB₁ in concentrations exceeding the NAFDAC recommended level for AFB₁ in food and reaching up to 2824 μg/kg. There was a weak positive correlation (r = 0.32, p = 0.03) for the relationship between the incidence of toxigenic strains in the samples and AFB₁ concentration. The consumer awareness data showed that 64% of the respondents consumed peanut cake; majority of who are youth of economic and reproductive age. Eighty-five percent of the consumers lacked awareness of aflatoxin contamination in the snack and possible health risks associated with its ingestion.     

Investigations on the essential oil of Lippia rugosa from Cameroon for its potential use as antifungal agent against Aspergillus flavus Link ex. Fries

Lippia rugosa essential oil was tested for its effectiveness against Aspergillus flavus on artificial growth media. The chemical composition of the oil was determined by gas chromatography–mass spectrometry (GC–MS). Geraniol (51.5%), nerol (18.6%) and geranial (10.4%) were the main components of Lippia oil. After 8 days of incubation on essential oil supplemented medium, mycelium growth of A. flavus was totally inhibited by 1000 mg l^?1 of L. rugosa essential oil. The effect of essential oil on aflatoxin B₁ synthesis was evaluated in SMKY broth. The medium supplemented with different essential oil concentrations, was inoculated with A. flavus mycelium and incubated at 25 °C. After 2, 4, 6 and 8 days, aflatoxin B₁ (AFB₁) was quantified in the supernatant using Enzyme Linked Immuno-Sorbent Assay (ELISA). Results showed that aflatoxin B₁ synthesis was inhibited by 1000 mg l^?1 of L. rugosa essential oil after 8 days of incubation. The effect of the EO on the H⁺-ATPase pumping membrane was also evaluated in the presence of several concentrations of oil (200–2000 mg l^?1) by monitoring glucose-induced acidification of the external medium. L. rugosa essential oil at the concentration of 2000 mg l^?1 completely inhibited the activity of this enzyme. These data suggest that the essential oil of L. rugosa is a fungicidal for A. flavus and its possible cellular target include the H⁺-ATPase.Results obtained in the present study indicate the possibility of exploiting Lippia rugosa essential oil in the fight against strains of A. flavus responsible for biodeterioration of stored foods products.     

Aflatoxin B1 production by Aspergillus parasiticus and strains of Aspergillus section Nigri in currants of Greek origin

Aflatoxin B₁ (AFB₁) mostly produced by Aspergillus flavus and Aspergillus parasiticus, is an extremely toxic and carcinogenic metabolite. Currants are used in the Mediterranean diet as a food with antioxidant properties. Four strains of Aspergillus section Nigri have been isolated from currants originated from Crete and Corinth. In this study AFB₁ production by A. parasiticus and the four strains of Aspergillus section Nigri in Cretan and Corinthian currants (Vitis vinifera L.) is investigated. AFB₁ determination was performed by HPLC–FID. Results revealed that the four strains Aspergillus section Nigri, as well as the aflatoxigenic strain A. parasiticus produced AFB₁ (0.0052–1.31 μg AFB₁ 15 g⁻¹, corresponding to 0.0003–0.087 μg AFB₁ g⁻¹) in both type of currants (Cretan and Corinthian) on the 12th day of observation. Moreover, AFB₁ production, by A. parasiticus in the synthetic Yeast Extract Sucrose (YES) medium was also studied. The ability of AFB₁ production has been affected by the special characteristics of each isolate and the currants substrate.     

Influence of cold plasma on fungal growth and aflatoxins production on groundnuts

In this study, influence of cold plasma on the Aspergillus parasiticus and Aspergillus flavus growth and aflatoxins productions have been investigated as an alternative to chemical free and thermal treatments. Artificially inoculated fungal species onto the groundnuts were treated with air plasma at 40 W and 60 W power levels at various time intervals. After the plasma treatment, inactivation of A. parasiticus and A. flavus was examined and the concentration of aflatoxins produced was analyzed using high performance liquid chromatography. There is 97.9% and 99.3% reduction in the growth of A. parasiticus and A. flavus respectively, when treated at 60 W powers. Electron microscopy was also used to study the effect of cold plasma on the cellular membranes of the spores. Results showed complete disintegration of fungal spore membrane due to electroporation and etching caused by the reactive species of plasma. In 40 W 15 min and 60 W 12 min plasma treated samples more than 70% and 90% reduction in aflatoxin B1 content was observed. These results suggest that cold plasma may be considered as alternative methods for disinfestation of foods due to its strong potential for microbial inactivation.     

Aflatoxins contamination in chilli samples from Pakistan

The aim of this study was to prioritise disease and pest constraints in chilli by highlighting aflatoxin concentrations to assist local farmers in control. All samples contained aflatoxin B₁ and high levels were obtained from all ground samples. A direct relationship was observed between aflatoxin B₁ and aflatoxin B₂ concentrations. There was no relation between aflatoxin and Aspergillus flavus detection. Chilli production in Pakistan may be heavily constrained by aflatoxin contamination. Simply removing A. flavus may be insufficient for control. Aflatoxins from chilli may be a threat to the health of populations and a constraint on development in Pakistan.     

The potential of some spice essential oils in the control of A. parasiticus CFR 223 and aflatoxin production

Essential oils of sweet basil (Ocimum basilicum), cassia (Cinnamomum cassia), coriander (Coriandrum sativum) and bay leaf (Laurus nobilis) at 1–5% (v/v) concentration in palm kernel broth inoculated with spore suspension (10⁶/ml) of Aspergillus parasiticus CFR 223 were evaluated for their potential in the control of aflatoxigenic fungus A. parasiticus CFR 223 and aflatoxin production. Healthy sorghum grains (120/treatment) immersed in the oils and distributed in three petri dishes with wet cotton wool were also inoculated with spore suspension (10⁶/ml) of A. parasiticus CFR 223 and assayed for grain protection. Sweet basil oil at optimal protective dosage of 5% (v/v) was fungistatic on A. parasiticus CFR 223 and aflatoxins produced in vitro and on fungal development on sorghum grains (P ⩽ 0.05) with a residual effect that lasted for 32 days. In contrast, oils of cassia and bay leaf stimulated the mycelia growth of the fungus in vitro but reduced the aflatoxin concentration (B₁ + G₁) of the fungus by 97.92% and 55.21% respectively, while coriander oil did not have any effect on both the mycelia growth and aflatoxin content of the fungus. The combination of cassia and sweet basil oils at half their optimal protective dosages (2.5% v/v) completely inhibited the growth of the fungus. The feasibility of implementing the results of this study to control aflatoxins was examined by the addition of whole and ground dry basil leaves at 5% and 10% (w/w), respectively, to 10 g sorghum, groundnut, maize and melon seed after 35 days storage period. It was found that the addition of whole and ground basil leaves markedly reduced aflatoxin contamination; however, 10% (w/w) of whole leaves was more effective as the reduction in aflatoxin was between 89.05% and 91%.The findings showed that aflatoxins can be controlled by co-storing whole sweet basil leaves with aflatoxin infected foods. The economic value of the study lies in the simplified technique for control of aflatoxin contamination in agricultural products and the benefits derivable from the use of local resources.     

Fungal and aflatoxin contamination of marketed spices

Fourteen spice samples were collected from local markets in Doha, Qatar, during 2012, and were surveyed for the presence of potentially harmful mycoflora and for contamination with aflatoxins B₁, B₂, G₁, and G₂ by high-performance liquid chromatography (HPLC). Among the tested spice samples, chili powder showed the highest presence of fungal propagules, while ginger, curry and garlic samples did not present any fungal contamination. A total of 120 isolates, mostly belonging to Aspergillus and Penicillium genera, were collected and 33 representative species were identified by amplification and sequencing of the internal transcribed spacer (ITS) region. Aspergillus flavus, Aspergillus nomius and Aspergillus niger were the most dominant. Thirty-seven Aspergillus strains were screened for their potential to produce aflatoxins using biochemical and molecular tools: only 9 A. flavus strains showed both fluorescence and amplification with all the three primers targeting aflP, aflM and aflR genes. Aflatoxins were detected in five spices (black pepper, chili, tandoori masala. turmeric and garam masala), and with the exception of garam masala, the tested samples of turmeric, black pepper, tandoori masala and chili powder exceeded B₁ and/or total aflatoxin maximum levels. Our results demonstrate the potential for mycotoxin biosynthesis by fungi contaminating imported spice products.     

Aflatoxin contamination of dried red chilies: Contrasts between the United States and Nigeria,two markets differing in regulation enforcement

Dried red chilies are among the world’s most consumed spices. From farm to fork, chilies go through cropping, harvest, drying, processing and storage. Chilies are susceptible to infection by aflatoxin producing fungi and subsequent contamination by aflatoxins at every stage. Aflatoxins are highly regulated, hepatotoxic carcinogens produced by fungi in Aspergillus section Flavi. The current study examined prevalence of aflatoxin B₁ (AFB₁) in chilies from markets across the United States (US) and Nigeria, and determined predisposition of chilies to aflatoxins post-harvest. Aflatoxin B₁ was detected in 64% chilies from US markets (n = 169), and 93% of Nigerian chilies (n = 55) with a commercial lateral flow assay (Limit of Detection = 2 μg/kg). Two percent of US samples exceeded the aflatoxin regulatory limit of 20 μg/kg, while the highest concentration detected was 94.9 μg/kg. Aspergillus spp. could be recovered only from 40% of samples from the US, and aflatoxin levels did not correlate with quantities of Aspergillus section Flavi (Colony Forming Units g⁻¹), suggesting fungi associated with chilies in US markets were killed during processing. Both average AFB₁ concentrations and fungal quantities were significantly higher (p < 0.01) in Nigerian chilies. The most contaminated sample contained 156 μg/kg AFB₁. Aflatoxin concentrations in Nigerian chilies increased as an exponential function of the quantities of Aspergillus section Flavi (r² = 0.76). Results indicate that high rates of chili consumption may be associated with unacceptable aflatoxin exposure.     

Inactivation of aflatoxigenic fungi (Aspergillus spp.) on granular food model,maize, in an atmospheric pressure fluidized bed plasma system

Atmospheric plasma provides the advantages of high microbial inactivation that can be performed under ambient conditions; therefore, it is regarded as a potential alternative to traditional food preservation methods. The present work presents the results of a critical study conducted on the efficiency of a non-thermal atmospheric pressure fluidized bed plasma (APFBP) system used for decontamination of maize. Maize grains that were artificially contaminated with Aspergillus flavus and Aspergillus parasiticus spores were treated in APFBP system for 1–5 min at two differently designed fluidized bed reactors with air and nitrogen. Results indicate maximum significant reductions of 5.48 and 5.20 log (cfu/g) in Aspergillus flavus and A. parasiticus after 5 min air plasma treatment. The native microbial flora of the maize grains decreased to more than 3 log after 3 min APFBP treatment, and no viable cells were counted. During the storage of plasma treated maize samples at 25 °C for 30 days, the Aspergillus spp. spores log reduction was maintained with no occurrence of re-growth. Overall, this study shows that plasma treatment has a fungicidal effect on A. flavus and A. parasiticus spores associated with alterations in spore surface morphology and loss of spore integrity. APFBP can inactivate aflatoxigenic spores on maize grains and could be optimized to improve the safety and quality of produce.     

Development of an aptasensor for the fast detection of Versicolorin A

Aflatoxins (AFs) are secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus. The molds may contribute to pre-harvest aflatoxin contamination of susceptible crops. For the customer and food producer, a predictive model for aflatoxin detection is very desirable. Versicolorin A (VerA), which is the first precursor in the pathway of aflatoxin B₁ (AFB₁) biosynthesis, shares similar toxic group with the furofuran structure in aflatoxin B₁. VerA exhibits a much lower teratogenic toxicity than AFB₁ and may be used as a predictive indicator for aflatoxin B₁ contamination of storage crops. Therefore, the development of a fast detection method for VerA is important. One of the randomly computer-generated aptamers of VerA was confirmed by isothermal titration calorimetry with K_d = 9.26 × 10⁻⁶ mol l⁻¹. In addition, a simple and sensitive label-free aptasensor was developed for the electrochemical detection of VerA. According to the results from differential pulse voltammetry (DPV), a linear relationship existed between the log conc. of VerA (ranged from 0.01 to 100 ng ml⁻¹) and the current (△I_p) with a limit of detection (LOD) of 10 pg ml⁻¹. The resulting aptasensor exhibited good reproducibility for detecting VerA and stability after storage for 15 days at 4 °C with acceptable anti-interference against ZEN, OTA, DON, and FB₁. When used in corn samples, the recoveries of VerA were determined to be in the range of 81.3%–104.4 %. Although with some intercross, result suggests that the obtained aptamer for VerA is potentially used as a sorbent for the preparation of solid-phase-extraction procedure to clean up food samples in conjunction with high-performance liquid chromatography analysis.     

Mycological quality and mycotoxin contamination of Sri Lankan peppers (Piper nigrum L.) and subsequent exposure assessment

The aim of the study was to characterize the toxigenic moulds and to screen different mycotoxins in peppers (Piper nigrum L.) of Sri Lankan origin. Aspergillus flavus, Aspergillus parasiticus, Aspergillus niger and Penicillium spp. were found to be the most dominant fungi. Characterization of the moulds was carried out in A. flavus and parasiticus agar (AFPA) and malt extract agar (MEA) in 77 black pepper (BP) and 11 white pepper (WP) samples. In total, 73% of the BP and 64% of the WP samples were contaminated with A. flavus and/or A. parasiticus (AfAp). A BP sample with water activity (a_w) 0.70 recorded the highest count of AfAp (4.3*10⁴ CFU/g). Moreover, 75% of the BP samples exceeded the safe a_w limit (0.65) set by the European Spice Association (ESA). The frequency of occurrence of A. niger in BP was 62% with counts up to 1.3*10³ CFU/g. Penicillium spp. were found in 61% and 55% of the BP and WP samples, respectively. In BP 94% of the samples had a Penicillium contamination below 10³ CFU/g. Other Aspergillus spp, found in peppers included, Aspergillus terrus, Aspergillus tamarii, Aspergillus candidus, Aspergillus penicilloides, Aspergillus sydowii and Aspergillus fumigatus. Mould counts in BP (10²–10⁴ CFU/g) were significantly higher than that of WP (<10² CFU/g). Apart from the occurrence of “classical mycotoxins” of spices, aflatoxins (<LOQ-18 μg/kg) and ochratoxin A (<LOQ-79 μg/kg), other toxins including fumonisin B1 (<LOQ-135 μg/kg), sterigmatocystin (<LOQ-49 μg/kg) and citrinin (<LOQ-112 μg/kg) were detected in peppers. In total, 63% of the BP samples were contaminated with at least one mycotoxin. Mycotoxin contamination in WP was significantly less compared to BP. The exposure to aflatoxins and ochratoxin A by consuming pepper remains harmless considering the existing pepper dietary intake data of the Sri Lankan population.     

Occurrence of aflatoxin B1 in food products derivable from ‘egusi’ melon seeds consumed in southwestern Nigeria

Aflatoxin, the toxic secondary metabolite of Aspergillus flavus and Aspergillus parasiticus has been considered as one of the most serious food safety problems in sub Saharan Africa. Egusi melon seeds is susceptible to these fungal infection during postharvest. Therefore, the content of aflatoxin B1 in three food products derivable from ‘egusi’ melon seeds ogiri’ (fermented melon seed condiment), ‘robo’ (melon ball snacks) and egusi soup destined for human consumption in Nigeria in 2005 and 2006 were determined. Aflatoxin B1 was analysed by thin layer chromatography (TLC) with fluorescent detection. The percentage of samples positive for aflatoxin B1 were for 25% robo, 31.8% for ogiri, and 19.5% for ‘egusi’ soup. Aflatoxin B1 ranged from 2.3 to 15.4 ppb in all samples. The overall mean levels of aflatoxin B1 were for 8.9 ppb for ogiri, 9.7 ppb for ‘robo’ and 7.2 ppb for ‘egusi soup’. All positive melon seeds derived food products analysed in this study contained aflatoxin B1 at concentrations much lower than the 20 ppb permissible limit recommended in Nigerian food and suggests that melon seed derived foods present less risk by human exposure to aflatoxin through their consumption. However, it is important to consider these levels in terms of their contribution to overall daily exposure to aflatoxin levels in food. This is the first report of aflatoxin determination in melon seed food products.     

Fungal contamination and aflatoxin B1 of ‘egusi’ melon seeds in Nigeria

One hundred and thirty seven samples of melon seeds (Colocynthis citrullus L.) from randomly selected farmers’ stores in the humid forest and Northern Guinea savanna of Nigeria were analysed for the incidence of diseased seeds, moisture content, associated moulds and levels of aflatoxin B₁ contamination. The proportion of diseased seeds ranged from 2.5 to 37.3% in the forest and 2.1 to 17.9% in the savanna, while the seed moisture content varied from 5.3 to 10.4%, and 4.6 to 9.5% respectively. All the samples contained moulds, with the two genera, Aspergillus and Penicillium predominating, while A. flavus had the highest species count. The other common fungal isolates in order of decreasing incidence were A. niger, P. citrinum, Botryodiplodia theobromae, Cladosporium sp and A. clavatus. Thin layer chromatography analysis showed that 32% in the forest and 21% samples in the savanna contained aflatoxin B₁ with mean levels of 14.8 μg/kg in the forest and 11.3 μg/kg in the savanna respectively. Significant positive correlations were found between number of aflatoxin B₁ positive samples and the percentage of A. flavus infected samples and between the levels of diseased seeds and the levels of aflatoxin B₁ contamination.     

Short wave infrared (SWIR) hyperspectral imaging technique for examination of aflatoxin B1 (AFB1) on corn kernels

Aflatoxins are toxic metabolites produced by the fungi Aspergillus flavus and Aspergillus parasiticus. They can contaminate a wide range of crops before harvest and during storage. Contaminated grains are associated with economic losses for cultivators as well as potential health hazards to both humans and animals. In this study, a short-wave infrared (SWIR) hyperspectral imaging technique was utilized to detect aflatoxin contamination on corn kernels. Corn samples were inoculated with four different aflatoxin B₁ (AFB1) concentrations (10, 100, 500 and 1000 μg/kg) while control samples were surface-disinfected with a PBS solution. Both infected and control samples were scanned with an SWIR hyperspectral system over the spectral range of 1100–1700 nm. A partial least squares discriminant analysis (PLS-DA) model was developed to categorize control and infected kernels and the highest overall classification accuracy yielded from the developed model was 96.9%. Spectral deviation was observed between the control and inoculated samples as the AFB1 concentrations increased. In addition, the contamination map generated with the PLS-DA model provided the visual appearance of infected samples. Our results suggest that SWIR hyperspectral imaging is a rapid, accurate, and non-destructive technique for the detection of toxic metabolites in grains and could be an alternative to manual techniques.     

Inactivation of aflatoxigenic fungi and the reduction of aflatoxin B1 in vitro and in situ using gamma irradiation

Detailed investigation on the effect of gamma (γ) irradiation on germination, sporulation, and growth of aflatoxigenic moulds (Aspergillus parasiticus 2999, Aspergillus flavus 305, and Aspergillus niger 388), as well as on the reduction of aflatoxin B₁ (AFB₁) level in artificially and naturally contaminated maize/feed samples was performed. The results of in vitro and in situ experiments with aflatoxigenic moulds demonstrated that 5 kGy-γ irradiation manages to prevent sporulation, germination and growth of the tested moulds both when in form of a pure and when in form of a mixed culture. In the feed samples artificially contaminated with AFB₁ (50 μg kg⁻¹) 5 kGy-γ irradiation reduced AFB₁ level by around 60%, while 10 kGy-dose reduce it for around 85%. Similarly, in feed samples spiked with AFB₁ in the concentrations of 100 μg kg⁻¹ 5 kGy-dose reduced the AFB₁ level by approximately 70%, while the dose of 10 kGy reduced it by approximately 90%. The experiments on naturally contaminated maize samples (n = 30) confirmed these observations; following a 5 kGy-irradiation, the overall mean AFB₁ reduction equalled to 69.8%, while the irradiation with a 10 kGy-dose achieved the overall mean toxin reduction of 94.5%. The obtained results indicate that γ irradiation can be used to prevent the growth of aflatoxigenic moulds and to reduce the AFB₁ levels in various goods intended for animal and human consumption, thus minimizing the animal and human exposure to this carcinogenic mycotoxin.     

Removal of Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 biofilms on stainless steel using scallop shell powder

Biofilms on steel surfaces containing Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 continue to threaten dairy and meat processors. In this study, the ability of scallop shell powder (SSP) to remove biofilms formed by these three pathogens on stainless steel plates was examined. Whey powder solution (WPS) and bench wash water (BWW) provided by dairy and meat factories, respectively, were inoculated with L. monocytogenes, S. aureus or E. coli O157:H7 (9 log₁₀ CFU/ml). Stainless steel plates (10 cm²) were placed in the inoculated fluids and incubated at 20 °C at 48 h to form biofilms. After drying and washing in sterile water, the plates were treated with 0.0, 0.25, or 0.50% (w/v) SSP slurries for 1, 5, or 10 min and then quantitatively examined for the three pathogens. Both 0.25 and 0.50% SSP reduced L. monocytogenes on the plates by 4 log CFU/cm² with a 1 min exposure to 0.50% SSP decreasing S. aureus by 5 logs CFU/cm². After 1 min in 0.25 and 0.50% SSP, E. coli O157:H7 populations in WPS and BWW biofilms decreased 4 and 6 log CFU/cm² and 3 and 5 log CFU/cm², respectively. Increasing the concentration of SSP led to significantly increased efficacy against the tested pathogens (P < 0.05). In conclusion, this study showed that SSP slurries could significantly reduce the numbers of L. monocytogenes, S. aureus and E. coli O157:H7 in biofilms on stainless steel surfaces.