Effect of two different roasting techniques on the Ochratoxin A (OTA) reduction in coffee beans (Coffea arabica)

The roasting of green coffee beans (Coffea arabica) artificially contaminated by Ochratoxin A (OTA) after inoculation with Aspergillus westerdijkiae was carried out with two different roasting techniques (Rotating Cylinder [RC] and Fluidized Bed [FB]). The green coffee beans were contaminated at two different toxin levels (L1 = 5.3 μg kg^?1 and L2 = 57.2 μg kg^?1). Different roasting points (light, medium, dark and very dark) were set according to the L¹ color coordinate. The cylinder roasting conditions were 0, 3, 6, 9, 12, and 15 min at 230 °C and the fluidized bed roasting conditions were 0, 0.9, 1.7, 2.6, 3.5 and 4.3 min, at 230 °C. The roasted beans were compared for their physical properties (bean swell and weight loss) as well as for their residual OTA content. The results indicated that the OTA reduction was similar for the two contamination levels: 95.1% and 97.2% with the rotating cylinder and 81.3% and 79.2% with the fluidized bed at the maximal roasting time. The OTA degradation kinetics differed between the two processes. The complete degradation of OTA within the limit of this study (230 °C) was not observed but the rotating cylinder roasting was the most efficient technical process for the OTA reduction in a commercial dark roasted coffee (88%).     

Pressurized liquid extraction coupled to liquid chromatography for the analysis of ochratoxin A in breakfast and infants cereals from Morocco

A sensitive and reliable method using pressurized liquid extraction (PLE) and liquid chromatography (LC) has been developed for the analysis of ochratoxin A (OTA) in breakfast and infants cereals. Influence of several extraction solvents that affect PLE efficiency was studied. The selected PLE operating method was: 10 g of sample was packed into 22 ml stainless-steel cell and OTA was extracted with acetonitrile/water (80:20) at 40 °C, 34 atm in one cycle of 5 min at 60% flush. The mean recovery of OTA was 82 ± 4 at fortification level of 3 ng/g OTA. The limit of quantification (LOQ) of OTA was 0.25 ng/g. The proposed method was successfully applied to the analysis of 68 samples of breakfast and infants cereals products collected from different supermarkets and pharmacies in Rabat. Results showed that all analyzed infant cereals were free of OTA contamination. However, four samples of breakfast cereals were contaminated with OTA. Levels of OTA in positive samples ranged between 5.1 and 224.6 ng/g. All positive samples (5.8% of total samples) were above the maximum level set by EU regulations for OTA in cereal derivatives products.     

Skin damage,high temperature and relative humidity as detrimental factors for Aspergillus carbonarius infection and ochratoxin A production in grapes

This study investigated the impact of skin damage on Aspergillus carbonarius colonization and ochratoxin A (OTA) production in grapes at different temperatures and relative humidity. Four ochratoxigenic A. carbonarius strains isolated from wine grapes were used to inoculate artificially damaged and undamaged table grapes. Grapes were stored at three levels of relative humidity (80%, 90% and 100%) and at two temperatures (20 and 30 °C). After seven days, the infection percentage of A. carbonarius was recorded and OTA accumulation in berries was analysed. Damaged grapes were more commonly infected and development of colonies was higher than in undamaged ones; consequently more OTA was detected in the former treatment. Temperature and relative humidity had significant influences on both infection and toxin content. The amount of OTA detected at 30 °C was higher than at 20 °C in most of the treatments. The highest relative humidity (100%) led to maximum amounts of OTA while no significant differences were found between 90% and 80% in the OTA content. The implementation of preventive measures in order to minimise berry damage in the field by controlling pathogenic fungi and insects during grape growing and removing visibly damaged grapes at harvest may significantly reduce OTA contamination in grapes.     

Superheated steam reduction of deoxynivalenol in naturally contaminated wheat kernels

Contamination of wheat with the Fusarium mycotoxin deoxynivalenol (DON) is a concern to the ethanol industry as it is stable during most processing operations and will be concentrated in the spent grains, which are potentially a valuable feedstock. Superheated steam at four processing temperatures (110, 135, 160, and 185 °C), three steam velocities (0.65, 1.3, and 1.5 m/s), and processing times of 2–15 min were used to treat wheat kernels naturally contaminated with DON to find the best processing parameters for the reduction of DON. A commercial enzyme-linked immunosorbent assay (ELISA) kit was used to determine DON levels in the wheat samples. Samples became increasingly toasted, displaying a brown color with increasing processing temperatures and times, and became friable after processing at temperatures of 160 and 185 °C. Only samples processed at 185 °C and 1.3 m/s exhibited any starch gelatinization. Significant (P < 0.05) reductions in DON levels were seen at 160 and 185 °C but were not generally seen at 110 and 135 °C and the effect of velocity was not significant (P > 0.05). Reductions of up to 52% were achieved at 185 °C and 6 min processing time and were due only to thermal degradation and not to solubilization and extraction.     

Deoxynivalenol reduction during the frying process of turnover pie covers

The effect of the frying process on deoxynivalenol contamination was evaluated. Deoxynivalenol naturally contaminated flour (1200 μg/kg) and fortified flour artificially contaminated (260 μg/kg) were used to prepare turnover pie dough covers. Frying was performed at three temperatures (169 °C, 205 °C and 243 °C) for different times. The final time for cooking at every temperature was established by measuring the colour during the frying process. Deoxynivalenol reduction was greater in the artificially contaminated samples (>66% at 169 °C, 43% at 205 °C and 38% at 243 °C). For the level of 1200 μg/kg, the average percentage of deoxynivalenol reduction, based on medians, was 28% when the dough covers were fried at 169 °C, 21% at 205 °C and 20% at 243 °C.     

Evaluating the combined effect of water activity,pH and temperature on ochratoxin A production by Aspergillus ochraceus and Aspergillus carbonarius οn culture medium and Corinth raisins

The objectives of this study were: (a) to evaluate the potential of the ELISA method in the determination of the produced OTA by Aspergillus ochraceus and Aspergillus carbonarius in malt extract agar (MEA) at different pH (3.9, 5.1, 5.9, 6.8), water activity (a_w) (0.87, 0.93, 0.99), and temperature (10, 15, 20, 25, 30, 40 °C) levels, providing a rapid screening for the optimum and marginal conditions of OTA production, (b) to comparatively evaluate the performance of ELISA and HPLC method, and (c) to evaluate the ability of A. ochraceus to produce OTA in rehydrated Corinth raisins during storage for 36 days. Two independent experiments were carried out to estimate OTA production on MEA and Corinth raisins. The produced OTA was evaluated qualitatively by the ELISA method and selected cases were verified by HPLC. The levels of OTA decreased with water activity, whereas pH seemed to have no specific effect. Furthermore, A. ochraceus produced maximum amounts of OTA on raisins at the 24th day of incubation, indicating that the endogenous microflora may restrictively affect OTA production. The knowledge of optimal and marginal levels of ecological factors in order to optimise post-harvest and storage of food products may significantly affect the production of OTA. Moreover, endogenous microflora of certain foodstuffs may cause OTA detoxification and consequently reduction of OTA levels; a fact that has to be taken into account in food commodities such as raisins, grapes, and wine.     

Hot air treatment for surface decontamination of table eggs

A hot air assay was set up for the surface decontamination of table eggs experimentally contaminated by Salmonella enterica serovar Enteritidis. A hot air apparatus was built and a treatment of two shots of 8 s at 600 °C with an interval of 30 s of cold air was chosen and applied on contaminated eggs. The S. Enteritidis load on eggshells as well as the quality traits of the egg components of 190 treated and 190 not treated eggs was investigated during 24 days of storage at 20 °C. Hot air treatment reduced the S. Enteritidis load on eggshells of up to 1.9 log. No significant changes to any of the quality traits tested were recorded. These results suggest the usefulness of hot air pasteurisation for the surface decontamination of table eggs.     

ELISA and HPLC analysis of ochratoxin A in red wines of Croatia

Ochratoxin A (OTA) contaminates wine all round the world, and its consumption may significantly increase human exposure to this toxin. In this study, enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC) for OTA analysis were tested on must and wine samples collected in Croatia. The results of ELISA and HPLC analysis of OTA in naturally contaminated red wines correlated well (r = 0.821), and the correlation was better at higher OTA concentrations. In contrast to HPLC, ELISA could not detect very low OTA concentrations. OTA concentrations in must (range 19–50 ng/l) were higher than in the wines (range 0–21 ng/l). No must samples showed the presence of Aspergillus carbonarius, which is a common OTA-producing mould affecting grapes.     

The development of an activated carbon from cherry stones and its use in the removal of ochratoxin A from red wine

The ability of activated carbon (AC) prepared from cherry stones (CS) by activation with H₃PO₄, ZnCl₂ or KOH to remove ochratoxin A (OTA) from two Italian red wines has been studied. AC was characterized in terms of texture and surface chemistry. OTA was analyzed by reversed-phase high-performance liquid chromatrography, using a fluorescence detector. The content of OTA in the starting wines is 7.38 and 2.36 μg/L. The adsorption of OTA is high only for one AC, which was prepared by KOH activation at 900 °C, using the 3:1 KOH:CS impregnation ratio. It possesses a large apparent surface area (S_BET = 1620 m²/g) and a high volume of large size macropores (1.84 cm³/g). It also contains narrow mesopores and intermediate size and wide micropores. Its content of acidic oxygen surface groups is low, whereas the content of basic groups is high (2.62 meq/g). The treatment of the wines with such an AC results in a decrease of the initial OTA content of more than 50%. However, the changes produced in the total polyphenolic index, color intensity, and hue are small (i.e. ～8%, ～5.5% and ～1.2%, respectively).     

Effect of organic acids,hydrogen peroxide and mild heat on inactivation of Escherichia coli O157:H7 on baby spinach

Minimally processed baby spinach contaminated with Escherichia coli O157:H7 has been associated with multiple outbreaks of foodborne illnesses recently. Chlorinated water is widely used to wash vegetables commercially, but this washing procedure has limited efficacy and can lead to the formation of carcinogenic substances. This study was conducted to determine the effects of organic acids and hydrogen peroxide alone and in binary combinations with or without mild heat (40 and 50 °C) on the inactivation of Escherichia coli O157:H7 on baby spinach. Baby spinach leaves were dip-inoculated with E. coli O157:H7 to a level of 6 log CFU/g and stored at 4 °C for 24 h before treatment. Individual washing solutions (1% and 2% lactic acid [LA], citric acid [CA], malic acid [MA], tartaric acid [TA], acetic acid [AA], hydrogen peroxide [H₂O₂] as well as binary combinations of LA, CA, MA and H₂O₂ at final concentrations of 1% were used to decontaminate spinach leaves at 22, 40 or 50 °C for 2–5 min to test their efficacy in reducing E. coli O157:H7. Chlorinated water (200 ppm free chlorine) decreased the population of E. coli O157:H7 on baby spinach by only 1.2–1.6 log CFU/g, which was not significantly different from DI water washing. Washing with 1% LA at 40 °C for 5 min was the most effective treatment achieving a 2.7 log reduction of E. coli O157:H7 which is significantly higher than chlorine washing. Washing with LA + CA or LA + HP at 40 °C for 5 min was equally effective against E. coli O157:H7, resulting in a 2.7 log reduction of E. coli O157:H7. The application of mild heat significantly enhanced the efficacy of washing solutions on the inactivation of E. coli O157:H7. There was, however, no significant difference between treatments at 40 °C for 5 min and 50 °C for 2 min. The results suggested that the use of organic acids in combination with mild heat can be a potential intervention to control E. coli O157:H7 on spinach.     

Degradation of aflatoxins in aqueous buffer in the presence of nucleophiles

This study compared the efficacies of lysine, glycine and methylamine to mediate aflatoxin destruction in heated aqueous phosphate buffers. A 3 × 3 × 4 factorial design (buffer pH, heating temperature and nucleophile) was followed. The buffer was artificially contaminated (“spiked”) with mixed aflatoxin standard and heated for specified time periods. After the heat treatment residual aflatoxins were determined using HPLC procedures.Aflatoxin reduction was influenced by the buffer pH and the pK_a of the added nucleophile. When phosphate buffer (at pH 9) was heated to 150 °C the degradation rate constants (k) aflatoxin in the presence of lysine, glycine and methylamine were 0.11, 0.09 and 0.08 min⁻¹ respectively. Addition of calcium chloride to the “spiked” buffer lowered aflatoxin reduction, but upon adding lysine or methylamine, aflatoxin reduction was restored to some extent. These findings demonstrate the potential capability of lysine to mediate aflatoxin reduction.     

Evaluation of chlorine,benzalkonium chloride and lactic acid as sanitizers for reducing Escherichia coli O157:H7 and Yersinia enterocolitica on fresh vegetables

The effectiveness in the assurance of fresh vegetable microbiological quality of wash solutions containing 200 ppm free chlorine, 0.1 mg/ml benzalkonium chloride, 0.2% and 1% lactic acid was assessed on Escherichia coli O157:H7 and Yersinia enterocolitica contaminated lettuce and tomatoes. Y. enterocolitica reduction on tomatoes (5.08, 4.77 and 4.21 log after 0.2% lactic acid, 200 ppm chlorine and 0.1 mg/ml benzalkonium chloride treatments, respectively) were significantly higher than those for Y. enterocolitica on lettuce and E. coli O157:H7 on both vegetables. Antimicrobial treatment effects on bacterial counts and product quality after subsequent 7 day storage (4 °C and 22 °C) were determined. No pathogens were found in natural microflora of fresh vegetables.     

Heat inactivation of Listeria innocua in broth and food products under non-isothermal conditions

The objective of this work was to study the effect of three linear temperature profiles (heating rates of 1.5, 1.8 and 2.6 °C/min, from 20 to 65 °C) on Listeria innocua inactivation in liquid medium. The inactivation was also analyzed in artificially contaminated parsley (heating rate of 1.8 °C/min) and throughout a frying process, using a pre-cooked frozen food as case study. Inactivation showed a sigmoidal behaviour and all data was fitted with a Gompertz-inspired model. Results demonstrated that, in liquid media, Listeria inactivation is influenced by the temperature profile used. As heating rate increases, the shoulder decreases and the tail effect disappears. If Listeria was in parsley, its heat resistance increased (for identical experimental conditions in broth). Besides model adequacy was proven in all studied situations, the heating rate affected parameters’ precision.     

Efficiency of high pressure treatment on inactivation of pathogenic microorganisms and enzymes in apple,orange, apricot and sour cherry juices

The purpose of this study was to investigate the effect of high hydrostatic pressure with a mild heat treatment on Staphylococcus aureus 485, Escherichia coli O157:H7 933 and Salmonella Enteritidis FDA in apple, orange, apricot and sour cherry juices. The effectiveness of the treatment on polyphenol oxidase activity in apple juice and pectinesterase activity in orange juice were also determined. An inoculum of microorganisms was completely inactivated at 350 MPa and 40 °C in 5 min. The residual polyphenol oxidase activity in apple juice after treatment at 450 MPa and 50 °C for 60 min was obtained as 9 ± 2.2%. The residual pectinesterase activity in orange juice after treatment at 450 MPa and 50 °C for 30 min was determined as approximately 7 ± 1.6%. It compares with 12 ± 0.2% at a treatment of 40 °C and 450 MPa for 60 min. Pressure resistant isoenzymes were thought to be responsible for the final residual activity. The inactivation is irreversible and the enzyme is not reactivated upon storage. High pressure processing constitutes an effective technology to inactivate the enzymes in fruit juices. Pressures higher than 400 MPa can be combined with mild heat (<50 °C) to accelerate enzyme inactivation.     

The growth,physiology and toxigenic potential of Bacillus cereus in cooked rice during storage temperature abuse

The effect of storage temperature abuse on the growth and physiology of Bacillus cereus in cooked rice was examined using plate counting and flow cytometry (FCM). Concurrently, toxigenic potential was measured through recording phosphatidylcholine-specific phospholipase C (PC-PLC) activity. Rice spiked with endospores was incubated at 4 °C, 10 °C or 18 °C for 6 days. Growth was not recorded at 4 °C, whereas >1.0 × 10⁶ CFU g^?1 were detected at 10 °C and 18 °C. PC-PLC activity was temperature dependent and was not destroyed by microwaving. FCM population profiling complemented plate count data, providing novel physiological data e.g. on the membrane integrity, redox or intracellular activities of cells over time during low temperature incubation.     

High hydrostatic pressure treatment applied to model cheeses made from cow’s milk inoculated with Staphylococcus aureus

Pasteurized milk was inoculated with two strains of Staphylococcus aureus (CECT4013 or ATCC13565) and used to elaborate soft-curd cheeses with approximately 7.5-log CFU/g of S. aureus. Cheeses were submitted to 10 min high hydrostatic pressure (HHP) treatments of 300, 400 or 500 MPa at 5 °C or 20 °C. Staphylococcus enterotoxin (SE) was evaluated in cheeses containing ATCC13565. Counts of S. aureus were measured after HHP treatment (day 1) and after 2, 15 and 30 days ripening at 8 °C. Inactivation increased with pressure and storage time, but was similar for both treatment temperatures. Maximum S. aureus reductions were achieved after 30 days ripening for samples treated at 500 MPa and 5 °C: 6.0 ± 0.1 and 4.7 ± 0.5-log CFU/g for CECT4013 and ATCC13565, respectively. However, SE was detected in all cheese samples containing ATCC13565 before and after HHP and after 30 days ripening.     

Comparison of aluminum thermal-death-time disks with a pilot-scale pasteurizer on the thermal inactivation of Escherichia coli K12 in apple cider

This study was conducted to compare thermal inactivation kinetics obtained using a pilot-scale pasteurizer and a bench-scale processing system. Pilot-scale pasteurizers are useful for product development, but comparisons on thermal inactivation kinetics with smaller scale systems are lacking. Using an Armfield pilot-scale pasteurizer and aluminum thermal-death-time (TDT) disks, the D-values and z-values of Escherichia coli K12 in apple cider were determined in the temperature range of 54–62 °C. Come-up times to 58 °C were also measured and were 35 and 61 s for the TDT disks and pasteurizer, respectively. The D-values from the TDT disks were 9.66, 4.01, 1.44 and 0.44 min at temperatures of 54, 56, 58, and 60 °C, respectively. The D-values from the pasteurizer were 3.48, 1.22, 0.10 and 0.05 min at temperatures of 56, 58, 60, and 62 °C, respectively. The z-values from the TDT disks and the pasteurizer were 4.68 and 3.60 °C, respectively. There was no significant (P > 0.05) difference in the D-values of the TDT disks and pasteurizer at 56 and 58 °C, while there was a significant (P < 0.05) difference in the D-value at 60 °C and in the z-value. This study revealed that the thermal inactivation kinetics obtained using bench scale TDT disks and an Armfield pilot-scale pasteurizer under certain conditions are similar. However, based on ease of use and other factors, TDT disks are preferable for acquiring thermal inactivation kinetics.     

Formation and diffusion mechanism of histamine in the muscle of tuna fish

This study investigates the formation and accumulation of histamine in the fresh meat of tuna fish. Under sterilized conditions, histamine was not detected in the muscles of Thunnus obesus (T. obesus) stored at 20 °C and 25 °C for 72 h (data not shown). And histamine formation and the diffusion mechanism was studied in T. obesus meat inoculated with two histamine-forming bacteria, Morganella morganii NBRC 3168 (M. morganii NBRC 3168) and Photobacterium phosphoreum NBRC 13896 (P. phosphoreum NBRC 13896) and stored at 20 °C and 25 °C for 3 days. The histamine level of the inoculated A1 point accumulated at a level above 4000 mg/kg in the T. obesus sample inoculated with M. morganii NBRC 3168 for 48 h. And the most level of the histamine in the remote B point was 2000 mg/kg at the time the sample was inoculated with M. morganii NBRC 3168 and stored at 25 °C. For the P. phosphoreum NBRC 13896 however, the level of histamine at the inoculated point A was 1800 mg/kg for 48 h when it was stored at 25 °C, while the highest level of histamine distant from the inoculum site B point was found to be approximately 1800 mg/kg. In contrast, when stored at 20 °C, histamine level was higher for the sample inoculated with P. phosphoreum NBRC 13896 than with M. morganii NBRC 3168. While histamine was diffused from the inoculation point in the M. morganii NBRC 3168 sample, it was diffused not only from the inoculated point, but also the remote area in the P. phosphoreum NBRC 13896 sample.     

Changes in the concentration of aflatoxin M1 during manufacture and storage of White Pickled cheese

In this study White Pickled cheese was produced from cow’s milk contaminated artificially with aflatoxin M₁ (AFM1) at two different levels, 1.5 and 3.5 μg/kg (ppb), and the effects of process stages on the AFM1 contents were investigated. Pasteurization at 72 °C for 2 min caused losses of AFM1 about 12% and 9%, respectively, in milk contaminated with 1.5 μg/kg AFM1, and 3.5 μg/l AFM1. These losses were found to be statistically significant (P < 0.01). After the cheese production, about 56% and 59% of total AFM1 remained in cheese–curd while about 32% of total AFM1 transferred to the whey for both 1.5 μg/kg and 3.5 μg/kg AFM1 contaminated milk. After 3-month storage in brine, AFM1 content of cheeses produced from 1.5 and 3.5 μg/kg AFM1 contaminated milks decreased by 2.9% and 2.8%, respectively. Changes in AFM1 content of cheese samples were found statistically insignificant (P > 0.05 and P > 0.01) for 3-month storage periods.     

Pre-processed bovine milk quality in Trinidad: Prevalence and characteristics of bacterial pathogens and occurrence of antimicrobial residues in milk from collection centres

The study was conducted to assess the impact of the changes in the milk collection system in Trinidad (from twice daily collection to once, introduction of chilling facilities to the collection centres and transportation of milk to the processing plant in insulated truck instead of in metal churns at ambient temperature) on the microbial load and antimicrobial residue quality of the milk as well as the temperature and pH of milk, using standard methods. The presence of antimicrobial residues was detected using the Delvo test kit. Of a total of 266 milk samples from churns, the mean ± sd temperature and log₁₀ ± sd TAPC per ml was 20.36 ± 7.91 °C and 6.3 ± 1.09 respectively. For 20 milk samples from the chillers, the mean temperature and log₁₀ ± sd TAPC per ml was 15.10 ± 2.73 °C and 7.04 ± 0.33 respectively compared with corresponding values for 36 samples collected from the truck, 11.64 ± 4.22 °C and 7.11 ± 0.62 respectively (P < 0.05; X²). The mean TAPC, staphylococcal and E. coli counts per ml of milk from churns were significantly (P < 0.05; X²) higher for milk at low temperature (0–20 °C) compared with milk at high temperature (>30 °C). Eight (4.2%) of 192 milk samples tested contained antimicrobial residues. Of 168 S. aureus isolates tested, 24 (14.3%) were enterotoxigenic while 53 (45.3%) of 117 isolates tested exhibited resistance to various antimicrobial agents while of 386 isolates of E. coli tested, 3 (0.8%) were O157 strain and 129 (64.5%) of 200 isolates exhibited resistance to antimicrobial agents. It was concluded that despite the changes, the microbial load of milk was still high suggesting poor sanitary practices at the farm level. The detection of antimicrobial residues agents coupled with toxigenic S. aureus and E. coli isolates could pose health hazards to consumers.