Assessment of multi-mycotoxin contamination throughout the supply chain of maize-based poultry feed from selected regions of Malaysia by LC-MS/MS.

The objective of this study was to examine the occurrence of multi-mycotoxin contamination throughout the supply chain of maize-based poultry feed. Different sampling points throughout the feed supply chain were selected from two companies that manufactured the poultry feed. A total of 51 samples, consisting of grain maize and maize-based poultry feeds, were collected. The samples were analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine the occurrence of multi-mycotoxin. The results revealed that 100% of samples throughout the maize-based poultry feed supply chain were spoiled with more than one mycotoxin. Fumonisin B1 (8.02-1,220 µg/kg) and fumonisin B2 (11.1-1,109 µg/kg) were the main mycotoxins detected at all sampling points throughout the feed supply chain. Zearalenone (ZEA) (6.63-7.50 µg/kg) was also detected in 11.7% (n = 6) (out of a total of 51) samples. As the supply chain progresses, a reduction in mycotoxin contamination was observed. Aflatoxins, ochratoxin A (OTA), deoxynivalenol (DON), HT-2, and T-2 toxin were not detected. The levels of mycotoxins detected throughout the supply chain were below the international regulatory limits, thus indicating the low risk of exposure to mycotoxins in maize-based poultry feed in Malaysia. Nevertheless, due to the presence of multiple ingredients in most food and feed, efforts to understand and address challenges associated with mycotoxins throughout the entire supply chain need to be more holistic to protect public health.

Simultaneous kinetics formation of heterocyclic amines and polycyclic aromatic hydrocarbons in phenylalanine model system.

A combination of chemical model system with kinetics study was used to investigate the simultaneous formation of heterocyclic amines (HCAs) and polycyclic aromatic hydrocarbons (PAHs). Heating a mixture of phenylalanine, creatinine, and glucose at a commonly practiced household cooking time and temperature successfully differentiated the rate formation (k) of HCAs and PAHs. The good fit suggested that the simultaneous formation was an endothermic bimolecular reaction, and followed the first-order model. The rate formation (k) of HCAs and PAHs significantly increased with increasing heating time and temperature. Only 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) showed degradation rate (k) at higher heating temperatures of 210 °C and 240 °C respectively. Increasing phenylalanine concentration increased the possibility of higher HCAs and PAHs formation. The activation energy (Ea) showed that heating phenylalanine mixture resulted in higher rate of formation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and benzo[b]fluoranthen (BbF).

Vitamin A and E Homologues Impacting the Fate of Acrylamide in Equimolar Asparagine-Glucose Model System.

This study aims to evaluate the influence of Vitamin A and E homologues toward acrylamide in equimolar asparagine-glucose model system. Vitamin A homologue as ß-carotene (BC) and five Vitamin E homologues, i.e., α-tocopherol (AT), δ-tocopherol (DT), α-tocotrienol (ATT), γ-tocotrienol (GTT), and δ-tocotrienol (DTT), were tested at different concentrations (1 and 10 µmol) and subjected to heating at 160 °C for 20 min before acrylamide quantification. At lower concentrations (1 µmol; 431, 403, 411 ppm, respectively), AT, DT, and GTT significantly increase acrylamide. Except for DT, enhancing concentration to 10 µmol (5370, 4310, 4250, 3970, and 4110 ppm, respectively) caused significant acrylamide formation. From linear regression model, acrylamide concentration demonstrated significant depreciation over concentration increase in AT (Beta = -83.0, R2 = 0.652, p ≤ 0.05) and DT (Beta = -71.6, R2 = 0.930, p ≤ 0.05). This study indicates that different Vitamin A and E homologue concentrations could determine their functionality either as antioxidants or pro-oxidants.

Selection of Vegetable Oils and Frying Cycles Influencing Acrylamide Formation in the Intermittently Fried Beef Nuggets.

This study aims to investigate the effect of different vegetable oils and frying cycles on acrylamide formation during the intermittent frying of beef nuggets. Different vegetable oils, palm olein (PO), red palm olein (RPO), sunflower oil (SFO), and soybean oil (SBO), were used for a total of 80 frying cycles. Oil was collected at every 16th frying cycle and analyzed for peroxide value (PV), p-anisidine value (p-AV), free fatty acid (FFA), total polar compound (TPC), polar compound fractions, and fatty acid composition (FAC). Total oxidation (TOTOX) value was calculated, and acrylamide content was quantified in the nuggets. Regardless of the oil type, PV, p-AV, and TOTOX initially increased but gradually decreased. However, FFA and TPC continued to develop across the 80 frying cycles. The C18:2/C16:0 remained almost unchanged in PO and RPO but dropped progressively in SFO and SBO. The lowest acrylamide content in fried products was observed in the PO, while the highest content was observed in RPO. Bivariate correlation analysis showed no significant (p ≤ 0.05) correlation between oil quality attributes and acrylamide concentration. The oil type but not the frying cycle significantly affected the acrylamide concentration in beef nuggets.

Prevalence of mycotoxigenic fungi and assessment of aflatoxin contamination: a multiple case study along the integrated corn-based poultry feed supply chain in Malaysia.

BACKGROUND: Corn, a main feed ingredient in the livestock industry, is one of the most susceptible crops to fungal infection and aflatoxin contamination. Livestock feeding on aflatoxin (AF)-contaminated feed have been shown to experience feed refusal, and decreased growth rate, milk production, and feed efficiency. In poultry, AF poisoning causes weight loss, poor feed efficiency, and reduced egg production and egg weight. The present work therefore aimed to determine the prevalence of mycotoxigenic fungi and the occurrence of AF contamination along the integrated corn-based poultry feed supply chain in Malaysia. A total of 51 samples were collected from different points along the feed supply chain from integrated poultry feed companies. The samples were subjected to mycological analyses (fungal isolation, enumeration, identification), and AFs were quantified by high-performance liquid chromatography equipped with a fluorescence detector (HPLC-FLD). RESULTS: Samples collected from sampling point 1 (company A) and sampling point 9 (company B) yielded the highest total fungal load (>log 4 CFU g-1 ). The prevalent fungal genera isolated were Aspergillus, Fusarium, and Penicillium spp. Aflatoxin B1 was detected in 8.3% of corn samples, and 7.4% of corn-based poultry feed samples along the feed supply chain, whereas AFs B2 , G1 , and G2 were not detected. CONCLUSION: The incidence of mycotoxigenic fungi along the integrated poultry feed supply chain warrant continuous monitoring of mycotoxin contamination to reduce the exposure risk of mycotoxin intake in poultry. © 2020 Society of Chemical Industry.

Phytopathogenic organisms and mycotoxigenic fungi: Why do we control one and neglect the other? A biological control perspective in Malaysia.

In this review, we present the current information on development and applications of biological control against phytopathogenic organisms as well as mycotoxigenic fungi in Malaysia as part of the integrated pest management (IPM) programs in a collective effort to achieve food security. Although the biological control of phytopathogenic organisms of economically important crops is well established and widely practiced in Malaysia with considerable success, the same cannot be said for mycotoxigenic fungi. This is surprising because the year round hot and humid Malaysian tropical climate is very conducive for the colonization of mycotoxigenic fungi and the potential contamination with mycotoxins. This suggests that less focus has been made on the control of mycotoxigenic species in the genera Aspergillus, Fusarium, and Penicillium in Malaysia, despite the food security and health implications of exposure to the mycotoxins produced by these species. At present, there is limited research in Malaysia related to biological control of the key mycotoxins, especially aflatoxins, Fusarium-related mycotoxins, and ochratoxin A, in key food and feed chains. The expected threats of climate change, its impacts on both plant physiology and the proliferation of mycotoxigenic fungi, and the contamination of food and feed commodities with mycotoxins, including the discovery of masked mycotoxins, will pose significant new global challenges that will impact on mycotoxin management strategies in food and feed crops worldwide. Future research, especially in Malaysia, should urgently focus on these challenges to develop IPM strategies that include biological control for minimizing mycotoxins in economically important food and feed chains for the benefit of ensuring food safety and food security under climate change scenarios.

Modelling the effect of temperature and water activity on the growth rate of Aspergillus flavus and aflatoxin production in peanut meal extract agar.

Aspergillus flavus is the predominant species that produce aflatoxins in stored peanuts under favourable conditions. This study aimed to describe the growth and aflatoxin production by two A. flavus strains isolated from imported raw peanuts and to model the effects of temperature and aw on their colony growth rate as a function of temperature and aw in Peanut Meal Extract Agar (PMEA). A full factorial design with seven aw levels (0.85-0.98 aw) and five temperature levels (20-40 °C) was used to investigate the growth and aflatoxin production. Colony diameter was measured daily for 28 days while AFB1 and total aflatoxin were determined on day 3, 7, 14, and 21. The maximum colony growth rate, µmax (mm/day) was estimated by using the primary model of Baranyi, and the µmax was then fitted to the secondary model; second-order polynomial and linear Arrhenius-Davey to describe the colony growth rate as a function of temperature and aw. The results indicated that both strains failed to grow at temperature of 20 °C with aw <0.94 and aw of 0.85 for all temperatures except 30 °C. The highest growth rate was observed at 30 °C, with 0.98 aw for both strains. The analysis of variance showed a significant effect of strain, temperature, and aw on the fungal growth and aflatoxin production (p < 0.05). Furthermore, both secondary models were in good agreement with the observed µmax. However, the polynomial model was found to be a better predictor of the experimental data. A similar pattern was observed in aflatoxin production but in a narrower range of temperature (25-35 °C) and aw (0.92-0.98 aw). The highest production of aflatoxins was observed on day 21 at 30 °C with the aw level of 0.98 for both strains. Overall, the current findings may help in improving the mycotoxin management and intervention strategies in peanuts, especially during storage.

Electrochemical determination of zearalenone using a label-free competitive aptasensor.

An electrochemical aptasensor is described for determination of the phytohormone of zearalenone (ZEA). The gold electrode was modified with ZEA via covalent attachment using cysteamine-hydrochloride and 1,4-phenylene diisocyanate linker. A truncated ZEA aptamer with a dissociation constant of 13.4 ± 2.1 nM was used in an aptasensor. The electrochemical property was investigated using square wave voltammetry for monitoring the change in the electron transfer using the ferro/ferricyanide system as redox probe. Under optimal experimental conditions, the response was best measured at a potential of 0.20 V (vs. Ag/AgCl). The signals depended on the competitive mechanism between the immobilised ZEA and free ZEA for the aptamer binding site. The aptasensor works in the range 0.01 to 1000 ng·mL-1 ZEA concentration, with a detection limit of 0.017 ng·mL-1. High degree of cross-reactivity with the other analogues of ZEA was observed, whereas none towards other mycotoxins. The aptasensor was further applied for the determination of ZEA in the extract of maize grain and showed good recovery percentages between 87 and 110%. Graphical abstract Schematic representation of the electrochemical determination of zearalenone based on indirect competitive assay. Step a Immobilisation of ZEA on the surface of gold electrode via covalent attachment, b competition for the ZEA aptamer binding site between immobilised and free ZEA, and c current signal of the binding event based on SWV technique.

Aspergillus section Flavi and Aflatoxins: Occurrence, Detection, and Identification in Raw Peanuts and Peanut-Based Products Along the Supply Chain.

Aflatoxin contamination in foods is a global concern as they are carcinogenic, teratogenic and mutagenic compounds. The aflatoxin-producing fungi, mainly from the Aspergillus section Flavi, are ubiquitous in nature and readily contaminate various food commodities, thereby affecting human's health. The incidence of aflatoxigenic Aspergillus spp. and aflatoxins in various types of food, especially raw peanuts and peanut-based products along the supply chain has been a concern particularly in countries having tropical and sub-tropical climate, including Malaysia. These climatic conditions naturally support the growth of Aspergillus section Flavi, especially A. flavus, particularly when raw peanuts and peanut-based products are stored under inappropriate conditions. Peanut supply chain generally consists of several major stakeholders which include the producers, collectors, exporters, importers, manufacturers, retailers and finally, the consumers. A thorough examination of the processes along the supply chain reveals that Aspergillus section Flavi and aflatoxins could occur at any step along the chain, from farm to table. Thus, this review aims to give an overview on the prevalence of Aspergillus section Flavi and the occurrence of aflatoxins in raw peanuts and peanut-based products, the impact of aflatoxins on global trade, and aflatoxin management in peanuts with a special focus on peanut supply chain in Malaysia. Furthermore, aflatoxin detection and quantification methods as well as the identification of Aspergillus section Flavi are also reviewed herein. This review could help to shed light to the researchers, peanut stakeholders and consumers on the risk of aflatoxin contamination in peanuts along the supply chain.

Molecular Characterisation of Aflatoxigenic and Non-Aflatoxigenic Strains of Aspergillus Section Flavi Isolated from Imported Peanuts along the Supply Chain in Malaysia.

Peanuts are widely consumed in many local dishes in southeast Asian countries, especially in Malaysia which is one of the major peanut-importing countries in this region. Therefore, Aspergillus spp. and aflatoxin contamination in peanuts during storage are becoming major concerns due to the tropical weather in this region that favours the growth of aflatoxigenic fungi. The present study thus aimed to molecularly identify and characterise the Aspergillus section Flavi isolated from imported peanuts in Malaysia. The internal transcribed spacer (ITS) and ß-tubulin sequences were used to confirm the species and determine the phylogenetic relationship among the isolates, while aflatoxin biosynthesis genes (aflR, aflP (omtA), aflD (nor-1), aflM (ver-1), and pksA) were targeted in a multiplex PCR to determine the toxigenic potential. A total of 76 and one isolates were confirmed as A. flavus and A. tamarii, respectively. The Maximum Likelihood (ML) phylogenetic tree resolved the species into two different clades in which all A. flavus (both aflatoxigenic and non-aflatoxigenic) were grouped in the same clade and A. tamarii was grouped in a different clade. The aflatoxin biosynthesis genes were detected in all aflatoxigenic A. flavus while the non-aflatoxigenic A. flavus failed to amplify at least one of the genes. The results indicated that both aflatoxigenic and non-aflatoxigenic A. flavus could survive in imported peanuts and, thus, appropriate storage conditions preferably with low temperature should be considered to avoid the re-emergence of aflatoxigenic A. flavus and the subsequent aflatoxin production in peanuts during storage.

The effects of different packaging materials, temperatures and water activities to control aflatoxin B1 production by Aspergillus flavus and A. parasiticus in stored peanuts.

Aflatoxins (AFs) are secondary metabolites produced by aflatoxigenic strains of Aspergillus flavus and A. parasiticus, the most toxic being aflatoxin B1 (AFB1). The purpose of the present work was to investigate the effects of industrial-grade packaging materials (low-density polyethylene, polypropylene, polyethylene-laminated aluminium); temperatures (25 °C, 30 °C); and water activities (0.74 a w, 0.85 a w) on AFB1 production by A. flavus and A. parasiticus in stored peanut kernels. Commercially-obtained samples were segregated into packaging materials, separately inoculated with the aflatoxigenic Aspergillus spp., and stored for 1 month under various °C + a w regimes. AFB1 production was quantified by high performance liquid chromatography with fluorescence detector (HPLC-FLD). For A. flavus in PELA, no AFB1 was detected (100% reduction) at 25 °C for both a w tested. For A. parasiticus in PELA, no AFB1 was detected at 25 °C (0.85 a w) and 30 °C (0.74 a w). Highest concentration of AFB1 was detected in LDPE for both A. flavus (46.41 ppb) and A. parasiticus (414.42 ppb), followed by PP (A. flavus 24.29 ppb; A. parasiticus 386.73 ppb). In conclusion, storing peanut kernels in PELA in a dry place at room temperature has been demonstrated as an adequate and inexpensive method in inhibiting growth of Aspergillus spp. and lowering AFB1 contamination in peanuts.

Heavy Metals in Selected Vegetables from Markets of Faisalabad, Pakistan.

Two hundred ten samples of selected vegetables (okra, pumpkin, tomato, potato, eggplant, spinach, and cabbage) from Faisalabad, Pakistan, were analyzed for the analysis of heavy metals: cadmium (Cd), lead (Pb), arsenic (As), and mercury (Hg). Inductively coupled plasma optical emission spectrometry was used for the analysis of heavy metals. The mean levels of Cd, Pb, As, and Hg were 0.24, 2.23, 0.58, and 7.98 mg/kg, respectively. The samples with Cd (27%), Pb (50%), and Hg (63%) exceeded the maximum residual levels set by the European Commission. The mean levels of heavy metals found in the current study are high and may pose significant health concerns for consumers. Furthermore, considerable attention should be paid to implement comprehensive monitoring and regulations.

Effect of Fat-Soluble Anti-oxidants in Vegetable Oils on Acrylamide Concentrations During Deep-Fat Frying of French Fries.

BACKGROUND: This study attempted to evaluate the effect of fat-soluble anti-oxidants in vegetable oils on acrylamide during the deep-fat frying of French fries. METHODS: Three vegetable oils with different fat-soluble anti-oxidant contents were selected and par-fried potato strips were fried in these oils. Acrylamide in the French fries at different frying times (at 180 °C) and over 10 consecutive frying sessions were measured. The anti-oxidant contents and quality degradation of oils were monitored before and after the 5th and 10th consecutive frying sessions. RESULTS: The effect of the fat-soluble anti-oxidants in red palm oil on the acrylamide was more apparent when a prolonged frying time was used for consecutive frying sessions than when different frying conditions were used. Using red palm oil, acrylamide concentration in French fries significantly dropped to the lowest level, at 524 ng g-1, after the 10th frying session. The ß-carotene content after the 10th frying session was the highest in red palm oil. CONCLUSION: The use of red palm oil for deep-fat frying French fries can be a mitigation strategy to reduce acrylamide formation, but further studies are necessary to investigate the influence of different types of fat-soluble anti-oxidants on the inhibition of acrylamide formation.

Reduction of sodium content in spicy soups using monosodium glutamate.

BACKGROUND: Excessive dietary sodium intake causes several diseases, such as hypertension, cardiovascular and renal disease, etc. Hence, reducing sodium intake has been highly recommended. In this study the effect of monosodium glutamate (MSG), as an umami substance, on saltiness and sodium reduction was investigated. METHODS AND RESULTS: The trained panellists were presented with basic spicy soups (curry chicken and chili chicken) containing different amounts of sodium chloride (NaCl) (0-1.2%) and MSG (0-1.2%). They tasted the optimum concentrations of NaCl and MSG for the two spicy soups and the overall acceptability were 0.8% and 0.7%, respectively. There was no significant effect of spiciness level on the saltiness and umami taste of both soups. The optimum levels of combined NaCl and MSG for overall acceptance in the chili and curry soups were 0.3% and 0.7%, respectively. The results showed that with the addition of MSG, it is possible to reduce sodium intake without changing the overall acceptability of the spicy soup. A 32.5% reduction in sodium level is made feasible by adding 0.7% MSG to the spicy soups. CONCLUSIONS: This study suggests that low-sodium soups can be developed by the addition of appropriate amounts of MSG, while maintaining the acceptability of the spicy soups. It was also proven that it is feasible to reduce sodium intake by replacing NaCl with MSG.

A Limited Survey of Aflatoxins and Zearalenone in Feed and Feed Ingredients from Pakistan.

This work presents current information on the presence of aflatoxins (AFs) and zearalenone (ZEN) in feed and feed ingredients from Punjab, Pakistan. The 105 samples tested were concentrated feed, i.e., cotton seed meal (18 samples) and soybean meal (14), and feed ingredients, i.e., crushed corn (17), crushed wheat (15), barley (17). and poultry feed (24). Samples were analyzed using high-performance liquid chromatography equipped with a fluorescence detector. Analysis revealed that 69 of 105 samples were contaminated with AFs, and the highest mean concentrations of AFB1 (6.20 µg/kg) and total AFs (9.30 µg/kg) were found in poultry feed samples. The mean total AF concentrations ranged from the limit of quantification to 165.5 µg/kg. However, 75 of the 105 samples were positive for ZEN. The highest mean concentration (19.45 µg/kg) was found in poultry feed samples. The mean ZEN concentrations were 0.15 to 145.30 µg/kg. The prevalence of AFs and ZEN was high in feed and feed ingredients and needs urgent attention.

Effect of roasting conditions on the browning intensity and structural changes in jackfruit (Artocarpus hetrophyllus) seeds.

Central composite rotatable design (CCRD) was used to optimize the settings for the roasting conditions of jackfruit (Artocapus hetrophyllus) seed (JFS). The response variables studied were; color attributes L*, a*, and b*, browning intensity, and fracturability. The colors L*, a*, b* and browning intensity were well predicted by a second-order polynomial model. Fracturability was predicted by a first-order polynomial. The determination coefficients for colors L*, a*, b*, browning intensity, and fracturability were 0.81, 0.96, 0.93, 0.92, and 0.74 respectively. The fitted models were checked for adequacy using analysis of variance (ANOVA). The optimum roasting conditions were established at a temperature of 153.36 °C, 34.36 min, and pH of 6.34 with composite desirability value of 0.95. Micro-structural studies of both raw and roasted JFS at different roasting levels (i.e., low, medium, and high) were also investigated using scanning electron microscope (SEM). JFS starch granules fell in the B-type category with semi-oval to bell-shaped granules (5-9 µm in diameter). In addition, Fourier Transform Infrared analysis was carried out on both raw and roasted JFS. The IR spectra was in the 4000-1000 cm(-1) region which is described by five main modes; O-H, C-H, C = O, (C-H) CH3, and C-O.

Modeling growth rate and assessing aflatoxins production by Aspergillus flavus as a function of water activity and temperature on polished and brown rice.

UNLABELLED: The aim of this study was to model the radial growth rate and to assess aflatoxin production by Aspergillus flavus as a function of water activity (a(w) 0.82 to 0.92) and temperature (12 to 42 °C) on polished and brown rice. The growth of the fungi, expressed as colony diameter (mm) was measured daily, and the aflatoxins were analyzed using HPLC with a fluorescence detector. The growth rates were estimated using the primary model of Baranyi, which describes the change in colony radius as a function of time. Total of 2 secondary models were used to describe the combined effects of a(w) and temperature on the growth rates. The models were validated using independent experimental data. Linear Arrhenius-Davey model proved to be the best predictor of A. flavus growth rates on polished and brown rice followed by polynomial model. The estimated optimal growth temperature was around 30 °C. A. flavus growth and aflatoxins were not detected at 0.82 a(w) on polished rice while growth and aflatoxins were detected at this a(w) between 25 and 35 °C on brown rice. The highest amounts of toxins were formed at the highest a(w) values (0.90 to 0.92) at a temperature of 20 °C after 21 d of incubation on both types of rice. Nevertheless, the consistencies of toxin production within a wider range of a(w) values occurred between 25 to 30 °C. Brown rice seems to support A. flavus growth and aflatoxin production more than the polished rice. PRACTICAL APPLICATION: The developed models can be used to estimate to what extent the change in grain ecosystem conditions affect the storage stability and safety of grains without the need for running long-standing storage study. By monitoring the intergranular relative humidity and temperature at different locations in the storage facility and inputting these data into the models, it is directly possible to assess either the conditions are conductive for the growth of A. flavus or aflatoxin production.