Natural occurrence of mycotoxins in cereals and spices commercialized in Morocco

Sixty samples of cereals (20 of corn, 20 of barley, and 20 of wheat) and 55 samples of spices (14 of paprika, 12 of ginger, 14 of cumin, and 15 of pepper) purchased from popular markets of Rabat and Salé in Morocco were analyzed for mycotoxins.Cereals samples were all analyzed for ochratoxin A (OTA). The average levels of contamination were 1.08, 0.42, and 0.17 μg/kg for corn, wheat, and barley, respectively. Samples of corn were also analyzed for zearalenone (ZEA) and fumonisin B₁ (FB₁) the average contaminations were 14 and 1930 μg/kg, respectively. Co-occurrence of OTA, FB₁, and ZEA was also checked. Spices samples were analyzed only for aflatoxins (AFs) and the average contaminations found for aflatoxin B₁ (AFB₁) were 0.09, 0.63, 2.88 and 0.03 μg/kg for black pepper, ginger, red paprika and cumin, respectively. The higher level of contamination was found in red paprika (9.68 μg/kg).The present report is the first one ever drafted on the natural co-occurrence of OTA, FB₁ and ZEA in cereals and on the occurrence of AFs in spices from Morocco.     

Use of gamma irradiation for inactivation of pathogens inoculated into Kimbab,steamed rice rolled by dried laver

The effect of gamma irradiation for inactivating the pathogens inoculated into the ready-to-eat Kimbab, steamed rice rolled by dried laver, was investigated. The pathogens used were Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus and Listeria ivanovii which are important for public health. The growth of four test organisms inoculated (about 10⁶–10⁷ CFU/g) into the Kimbab were sustained by an irradiation treatment during 24 h of storage regardless of the temperature at 10, 20 and 30 °C. Four pathogen inoculated into Kimbab decreased 2–3 log CFU/g by 1 kGy treatment and was not detected after 3 kGy. The D₁₀ value of pathogens inoculated into the Kimbab were 0.31–0.44 kGy among the four organisms. This study indicated that a low dose irradiation can maintain microbial safety for ready-to-eat Kimbab, steamed rice rolled by dried laver.     

Estimation of dietary intakes of fumonisins B1 and B2 from conventional and organic corn

The dietary intakes of fumonisins from 60 samples of conventional and organic corn were assessed. A 13.3% of the conventional corn samples contained fumonisin B₁ and B₂ at mean levels of 43 and 22 ng/g, respectively, while 10% of the organic corn samples contained fumonisins at somewhat lower levels of 35 ng/g (FB₁) and 19 ng/g (FB₂). Overall, the fumonisin levels in the corn samples were much lower than the maximum level of 2000 ng/g (as the sum of FB₁ and FB₂) proposed for unprocessed maize in a recent EU regulation. The fumonisins present in conventional and organic maize are estimated to contribute with very low percentages of 0.21% and 0.17%, respectively, to the level considered at risk for human health. Based on the data exposed in this paper, the farming system is probably not of decisive importance for the final contamination of agricultural products with these mycotoxins.     

Radiation sensitivity of selected pathogens in ice cream

Microbial contamination was investigated using ice creams with a vanilla, chocolate, and strawberry flavor commercially available in Korea. Radiation sensitivity of the food-borne pathogens was also determined by an inoculation test. Food-borne pathogens used were Listeria ivanovii, Escherichia coli, and Salmonella typhimurium. Total aerobic bacteria, moulds and yeasts, and coliforms in the ice creams ranged from 2 to 3 log CFU/g. Irradiation of 3 kGy was enough to inactivate the total aerobic bacteria for the vanilla ice cream but that of 5 kGy was needed for the chocolate or strawberry ice creams at a frozen condition (−20 °C). To inactivate (>log 6.5) the inoculated L. ivanovii, E. coli, and S. typhimurium into ice cream irradiation of 3, 1, and 0.1 kGy was needed, respectively. The D₁₀ value of L. ivanovii and E. coli was calculated as 0.71–0.77 and 0.28–0.38 kGy range for the ice cream with different flavors at −72 °C, respectively. The D₁₀ value of S. typhimurium could not be calculated in this study because even 0.1 kGy of irradiation reduced the number of S. typhimurium to undetected level. Results suggest that a low dose irradiation can improve the microbial quality and reduce the risk by the food-borne pathogens of ice cream, which has limited alternative sterilization methods due to the temperature characteristics of the 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.     

Microbial and quality evaluation of green peppers stored in biodegradable film packaging

The effects of polylactic acid (PLA) based biodegradable film packaging on the microbial and physicochemical quality of green peppers (Capsicum annuum L.) were compared to the effects of low-density polyethylene (LDPE) film packaging, and perforated LDPE film packaging. Each package containing green peppers was heat-sealed and stored for 7 days at 10 °C. The microbial levels (aerobic bacteria, coliform bacteria, and yeast and moulds) and physicochemical properties such as colour, weight loss, hardness, ascorbic acid concentration, O₂ and CO₂ concentrations, were monitored during storage. Results indicated that the physicochemical properties of colour, hardness, ascorbic acid concentration, and microbial levels (total aerobic bacteria, and moulds and yeasts) did not show remarkable changes during storage period. The microbial levels in coliform bacteria were increased by less than 1 log CFU/g (0.2 log CFU/g) in the biodegradable film packaging, 2.3 log CFU/g in LDPE film package, and less than 1 log CFU/g (0.9 log CFU/g) in the perforated LDPE film package, after 7 days storage period. The results suggest that the biodegradable film with higher water vapor permeability can be used to maintain the quality and sanitary conditions (protection from microbial and insect contamination) of freshly harvested green peppers in modified atmosphere packaging.     

Safe and shelf-stable natural casing using hurdle technology

Safe and shelf-stable natural casing were prepared using a combination of hurdles viz. reduced water activity, packaging and gamma irradiation. Washed lamb intestines were treated with common salt to reduce water activity to 0.80 ± 0.02, packed in polyethylene bags and subjected to gamma-irradiation (5 and 10 kGy). Control non-irradiated samples had high total viable counts (10⁶ CFU/g), aerobic spores (10³ CFU/g), spores of sulphite reducing clostridia (10³ CFU/g), potentially pathogenic bacteria such as staphylococci (10⁴ CFU/g) and coliforms (10² CFU/g). Treatment with gamma radiation resulted in a dose dependent reduction in counts of these microbes. A dose of 5 kGy was sufficient to reduce total viable counts by three log cycles; spore counts by two log cycles and completely eliminate staphylococci and coliforms. Samples subjected to a 10 kGy dose were devoid of any viable microbes. The reduced water activity of the product prevented growth of the microbes in natural casings during storage at room temperature. Sausages prepared using hurdle processed natural casing were examined for sensory and textural properties. It was observed that product acceptability and mechanical strength was not affected by radiation processing. Our studies indicated that shelf-stable and safe natural casing could be prepared using a combination of hurdles.     

Incidence of zearalenone and fumonisins in Bulgarian cereal production

Ninty-one small-grain cereals (wheat, barley, maize) collected during the 2007 harvest in Bulgaria were tested for zearalenone (ZON) and fumonisins contamination. Analytical methods based on immunoaffinity clean-up and detection by liquid chromatography was used after validation. Limits of detection for ZON in different matrices were below 4.0 μg/kg in barley and wheat, and slightly higher for maize (17.6 μg/kg). The limit of quantification for ZON was 12 μg/kg in barley and wheat, and 58.8 μg/kg in maize. Recovery values ranged between 84% and 105%. The occurrence of ZON in cereals were rather low and only single incidences was found – up to 148 μg/kg for maize and 36.6 μg/kg for other cereals. Fumonisins in maize have showed a widespread distribution (in 94.7% of tested samples). One of the tested samples was contaminated above the established maximum limits for unprocessed maize.     

Microbiological evaluation of fresh-cut organic vegetables produced in Zambia

This study was undertaken to assess the microbiological quality of fresh-cut organic vegetables produced in Zambia. Fresh-cut organic mixed vegetables and green beans produced in Zambia were analysed for aerobic plate counts, coliforms, Enterobacteriaceae, Escherichia coli, Bacillus cereus, Clostridium perfringens, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and yeast and mould counts. The study included 160 samples for most of the parameters. The vegetables were grown on farms meant primarily for the export market. The vegetables were treated/washed with 150 μg ml⁻¹ chlorine solution at the processing plant prior to sampling. The aerobic plate count ranged between 3 log₁₀ and 9.7 log₁₀ CFU/g, with the highest count recorded for green beans. The largest grouping (26.1%) of vegetable samples fell between 3 and 4 log₁₀ CFU/g. Coliform counts were between 1.0 log₁₀ and 7.7 log₁₀ CFU/g. The highest incidence level was 31.4% for total coliform counts between 3 log₁₀ and 4 log₁₀ CFU/g. E. coli was only detected on mixed vegetables in the range of 0.6 log₁₀ to 3 log₁₀ CFU/g, while Enterobacteriaceae counts ranged between 1.6 log₁₀ and 9.8 log₁₀ CFU/g with the highest counts being found on green beans. The highest incidence level was of 25.8% for counts within the same range as the aerobic plate counts. Yeast and mould counts showed the highest incidence level between 5 log₁₀ and 6 log₁₀ CFU/g with an overall range between 1.5 log₁₀ and 5.6 log₁₀ CFU/g. L. monocytogenes, Salmonella spp. and S. aureus were detected in 20%, 23.1% and 83.9% of samples, respectively . C. perfringens and B. cereus were not detected in any of the samples analysed.     

Inactivation of 3-strain cocktail pathogens inoculated into Bajirak jeotkal,salted, seasoned,and fermented short-necked clam (Tapes pilippinarum), by gamma and electron beam irradiation

The objective of this study was to determine the efficacy of gamma and electron beam irradiation of the food-borne pathogens including 3-strain cocktail of Listeria monocytogenes (ATCC 19114, 19115, and 19111), Staphylococcus aureus (ATCC 6538, 25923, and 29213), and Vibrio parahaemolyticus (ATCC 17802, 33844, and 27969) in Bajirak jeotkal (8% salt), salted, seasoned and fermented short-necked clam, commercially available in the market. Irradiation (0.5, 1, 2, and 5 kGy) significantly reduced the initial microbial level not only immediately after irradiation but also during storage at 10 °C for 4 weeks (P < 0.05). No viable cells were detected at 5 kGy of irradiation at a detection limit of 10¹ CFU/g. Gamma irradiation was more effective than electron beam irradiation, and yielded D₁₀ values of 0.64, 0.63, and 0.29 kGy for L. monocytogenes, S. aureus, and V. parahaemolyticus, and those of electron beam irradiation were 0.79, 0.81, and 0.36 kGy, respectively. Results suggest that a low dose irradiation can improve the microbial quality and reduce the risk by the food-borne pathogens of Bajirak jeotkal, which has limited alternative sterilization methods due to the temperature characteristics of the products. Furthermore, in practical application, the irradiation source should be considered to obtain an effective dose for decontamination.     

Effect of gamma radiation on reduction of mycotoxins in black pepper

Gamma ray was applied to reduce mycotoxins, i.e. ochratoxin A (OTA) and aflatoxins B₁, B₂, G₁ and G₂ (AFB₁, AFB₂, AFG₁ and AFG₂) in black pepper. Response surface methodology (RSM) was applied to evaluate the effect of dose of gamma ray ranging from 0 to 60 kGy and mycotoxin concentration ranging from 10 to 100 ng g⁻¹ on the mycotoxin reduction. The maximum reduction was found at 60 kGy which was 52%, 43%, 24%, 40% and 36% for OTA, AFB₁, AFB₂, AFG₁ and AFG₂, respectively. Results showed the gamma rays even at 60 kGy were not effective in completely destroying of ochratoxin and aflatoxins.     

Effect of gamma irradiation on Listeria ivanovii inoculated to iceberg lettuce stored at cold temperature

A low-dose gamma irradiation considerably reduced the total plate counts, psychrotrophic bacteria, lactic acid bacteria, and inoculated Listeria ivanovii on shredded iceberg lettuce. The total plate count of the lettuce irradiated at 1.0 kGy was reduced by 3.38 log₁₀ cfu/g on 0 storage day and to below the limit of detection (<2 log₁₀ cfu/g) as the cold storage was extended. Irradiation at 0.5 kGy effectively reduced the psychrotrophic bacterial counts and lactic acid bacterial counts in the lettuce to below the limit of detection (2 log₁₀ cfu/g). Irradiation at 1.0 kGy reduced L. ivanovii inoculated onto the shredded iceberg lettuce to below the limit of detection. The results showed that an irradiation at 1 kGy eliminates the bacterial contamination from the lettuce sample without any sensorial quality defect.     

Inactivation of a non-pathogenic strain of E. coli by ionising radiation

Ionising radiation is an effective method to reduce pathogenic E. coli O157:H7 in meat and poultry products. Radiation sensitivity of bacteria, however, depends on several factors. After applying an irradiation dose of 1 kGy to cultures of the non-pathogenic strain of E. coli, DSM 498, grown and irradiated in nutrient broth, reductions of 3–4 decimal units were achieved (D₁₀ = 0.27 kGy). If grown on minced turkey meat, however, reduction rates were lower (D₁₀ = 0.47 kGy). Even lower reduction rates were obtained during irradiation of frozen meat (D₁₀ = 0.72 kGy) compared to treatments at cooling temperatures (D₁₀ = 0.48 kGy). For data evaluation, both, first order reduction kinetics and the Weibull model were compared. The results emphasise the necessity to determine inactivation kinetics in food matrices of target extrinsic factors (e.g. temperature).     

Effect of γ irradiation on quality-maintaining of fresh-cut lettuce

The effects of irradiation on microorganisms and physiological quality of fresh-cut lettuce were evaluated during storage at 4 °C. The total bacterial counts on fresh-cut lettuce irradiated with 1.0 kGy were reduced by the order of 2.35 Log CFU/g, and the total coliform group were lowered to less than 30 MPN (most probably number)/100 g. The polyphenoloxidase activity of fresh-cut lettuce was significantly inhibited by irradiation. In addition, the loss of vitamin C of fresh-cut lettuce irradiated with 1.0 kGy was significantly (α = 0.05) lower than that of non-irradiated. The best treatment of maintaining quality of fresh-cut lettuce appeared to be 1.0 kGy irradiation.     

Detection of radiation-induced markers from parts of irradiated kiwifruits

Parts of irradiated kiwifruits were investigated for radiation-induced markers. The DNA comet assay on seeds showed that non-irradiated cells appeared as intact nuclei without tails, while irradiated cells displayed comets with long tails. Tail length of seeds increased significantly up to 1 kGy (p < 0.05), showing a positive correlation (R² = 0.9084) between the irradiation dose and tail length. The triplet signals from cellulose radicals observed in electron spin resonance (ESR) analysis on the core or flesh of kiwi enabled irradiated samples to be distinguished from the non-irradiated samples. The ESR signals increased in correlation with the irradiation doses with R² = 0.9071 in the core and R² = 0.9730 in flesh. Both methods were found suitable for the detection of irradiated kiwi during the six-week shelf-life period.     

Effects of gamma irradiation on the microbiological,nutritional, and sensory properties of fresh vegetable juice

The radiation pasteurization process was performed to improve the microbiological quality of fresh vegetable juice. Carrot and kale juice were irradiated and their microbiological, nutritional, and sensory properties were evaluated. The contaminating bacteria in the juices before irradiation ranged from 10⁶ to 10⁷ CFU/ml. All the aerobic and coliform bacteria in the carrot juice were eliminated by irradiation at a dose of 3 kGy, whereas about 10² CFU/ml of the bacteria survived in the kale juice irradiated at up to 5 kGy. However, the cells that survived from irradiation in the kale juice did not grow, whereas those of the non-irradiated samples reached 10⁹ CFU/ml after 3 days of storage at 10 °C. Amino acids were stable at up to 5 kGy of an irradiation. Radiation resulted in a dose-dependent reduction of the ascorbic acid content. However, the contents of the total ascorbic acid, including dehydroascorbic acid, were stable at up to 3 kGy of an irradiation. The sensory evaluation results immediately after irradiation were not different in any of the samples. At a 3-day storage, the sensory quality of the irradiated juice was adequate, while the quality of the non-irradiated control was deteriorated.     

A rapid FT-NIR method for estimation of aflatoxin B1 in red chili powder

The feasibility of measuring Aflatoxin B₁ (AFB₁) in red chili powder was investigated by using Fourier transform near-infrared (FT-NIR) spectroscopy in diffuse reflectance mode combined with appropriate chemometric techniques. Aflatoxin free chili powder samples were spiked with known amount of AFB₁ ranging from 15 to 500 μg/kg and used for calibration model building based on partial least squares (PLS) regression algorithm. Different spectral preprocessing methods were investigated and optimized based on the lowest values of root mean square error of cross validation (RMSECV). Spectral wavenumber range of 6900.3–4998.8 and 4902.3–3999.8 cm^?1 and straight line subtraction preprocessing technique predicted AFB₁ content with best accuracy with lowest RMSECV = 0.654% and maximum correlation coefficient for validation plots (R² = 96.7). The overall results demonstrate that FT-NIR spectroscopy can be used for rapid, non destructive quantification of Aflatoxin B₁ in red chili powder.     

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.     

Inhibition of Staphylococcus aureus on beef by nisin-containing modified alginate films and beads

Nisin, a bacteriocin, was immobilized into palmitoylated alginate-based films or in activated alginate beads. Sterile beef muscle slices or ground beef were inoculated with Staphylococcus aureus at a level of 10⁴ CFU/g. Sliced beef was then coated with palmitoylated alginate-based films containing 0, 500 or 1000 IU/mL of nisin. Also, ground beef was mixed with 0, 500 or 1000 IU/mL of nisin covalently linked to activated alginate beads in order to evaluate the effect of nisin concentration on S. aureus level. The content of S. aureus in beef was determined during storage at 4 °C. Results demonstrated that after 7 days of storage, a reduction of 0.91 and 1.86 log CFU/cm² was observed on sliced beef covered with film containing 500 or 1000 IU/mL of nisin, respectively. After 14 days of storage, when nisin solution (500 or 1000 IU/g) was mixed with ground beef, 2.2 and 2.81 log CFU/g reductions of S. aureus counts were respectively observed. However, when nisin (500 or 1000 IU/g) was linked into activated alginate beads, 1.77 and 1.93 log CFU/g reductions of S. aureus counts were respectively observed (P ⩽ 0.05). These results suggest that sterile, hydrophobic and biodegradable films or beads incorporating various amounts of nisin could be used efficiently to control the growth of pathogens or microorganisms responsible of spoilage at the surface of round beef or other meat products.     

Application of slightly acidic electrolyzed water as a potential non-thermal food sanitizer for decontamination of fresh ready-to-eat vegetables and sprouts

The sanitization efficacy of slightly acidic electrolyzed water (SAEW) against food pathogens on selected fresh ready-to-eat (RTE) vegetables and sprouts was evaluated and compared to sodium hypochlorite (NaOCl) solution. RTE vegetables and sprouts were dip-inoculated with Escherichia coli (E. coli) and Salmonella spp. and dip-treated with SAEW, NaOCl solution for 5 min. SAEW treatment significantly (p < 0.05) reduced the total aerobic mesophilic bacteria from Chinese celery, lettuce and daikon sprouts by 2.7, 2.5 and 2.45 log₁₀CFU/g, respectively relative to un-treated. Pathogens were significantly (p < 0.05) reduced from Chinese celery, lettuce and daikon sprouts by 2.7, 2.8 and 2.8 log₁₀CFU/g (E. coli) and 2.87, 2.91 and 2.91 log₁₀CFU/g (Salmonella spp.), respectively following a SAEW treatment. SAEW and NaOCl solution showed no significant sanitization difference (p > 0.05). Results demonstrate that SAEW at low chlorine concentration and a near neutral pH is a potential non-thermal food sanitizer that could represent an alternative to NaOCl solution and would reduce the amount of free chlorine used in fresh-cut vegetables industry, since the same microbial reduction as NaOCl solution is obtained.