Efficacy of Artabotrys odoratissimus oil as a plant based antimicrobial against storage fungi and aflatoxin B1 secretion

The essential oil of Artabotrys odoratissimus R.Br. was evaluated for antifungal activity against some storage fungi causing contamination of food stuffs. Minimum inhibitory concentration of the oil was found to be 750 μL L⁻¹ against Aspergillus flavus Link. It was found superior over different prevalent synthetic fungicides which inhibited the growth of A. flavus between 1000–5000 μL L⁻¹. The oil exhibited broad fungitoxic spectrum against fourteen different storage fungi. Aspergillus fumigatus was inhibited at 1000 μL L⁻¹ whereas Cladosporium cladosporioides , Curvularia lunata , Fusarium oxysporum , Helminthosporium oryzae , Macrophomina phaseolina , Microsporum gypseum , Mucor racemosus , Penicillium italicum , Pythium debaryanum , Rhizoctonia solani , Sclerotium rolfsii and Trichoderma viride at 500 μL L⁻¹. Aspergillus niger was found to be inhibited only 84.9% at 1000 μL L⁻¹. In addition, the oil showed significant efficacy in arresting aflatoxin B₁ secretion by the toxigenic strain (Navjot 4NSt) of A. flavus at 750 μL L⁻¹. The efficacy of A. odoratissimus oil as aflatoxin suppressor is being reported for the first time.     

Radical scavenging and antimicrobial activity of horsetail (Equisetum arvense L.) extracts

A reversed-phase high performance liquid chromatography (RP-HPLC) separation on C₈ column and quantitative method were developed to analyse hydroxyl derivatives of benzoic and cinnamic acid and flavonoids in horsetail ( Equisetum arvense L.) extracts. Total phenolic content of n -butanol, ethyl acetate and water extracts, determined by the Folin-Ciocalteu method, was 96.4, 26.4 and 15.4 mg g⁻¹ of dry extracts, respectively. The antioxidative activity of horsetail extracts was tested by measuring their ability to scavenge stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reactive hydroxyl radicals by electron spin resonance spectroscopy. The results demonstrated that the free radical scavenging activity (versus both DPPH and hydroxyl radicals) depended on the type and concentration of applied extracts; the highest DPPH (EC₅₀ = 0.65 mg mL⁻¹) and hydroxyl radical scavenging activities (EC₅₀ = 0.74 mg mL⁻¹) were obtained in the case of n -butanol extract. The radical scavenging activity of extracts significantly correlated with total phenolic content. The antimicrobial tests showed that ethyl acetate and n -butanol extracts inhibited the growth of tested bacteria.     

Essential Oil of Aegle marmelos as a Safe Plant-Based Antimicrobial Against Postharvest Microbial Infestations and Aflatoxin Contamination of Food Commodities

ABSTRACT:  The essential oil of  Aegle marmelos  L. Correa (Rutaceae) showed strong fungitoxicity against some storage fungi-causing contamination of foodstuffs. The oil also showed efficacy as aflatoxin suppressor at 500 μL/L as it completely arrested the aflatoxin B₁ production by the toxigenic strains (Navjot 4NSt and Saktiman 3NSt) of  Aspergillus flavus  Link. Keeping in view the side effects of synthetic fungicides,  A. marmelos  oil may be recommended as an antimicrobial of plant origin to enhance the shelf life of stored food commodities by controlling the fungal growth as well as aflatoxin secretion. This is the 1st report on aflatoxin B₁ inhibitory nature of this oil.  A. marmelos  oil may be recommended as a novel plant-based antimicrobial in food protection over synthetic preservatives, most of which are reported to incite environmental problems because of their nonbiodegradable nature and side effects on mammals. The LD₅₀ of  Aegle  oil was found to be 23659.93 mg/kg body weight in mice ( Mus musculus  L.) when administered for acute oral toxicity showing nonmammalian toxicity of the oil. GC-MS analysis of the oil found DL-Limonene to be major component.     

Antioxidant and toxicity tests of roasted noni (Morinda citrifolia) leaf infusion

The antioxidant properties and toxicity profile of roasted noni ( Morinda citrifolia L . ) leaf infusion were evaluated. The 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging activity was greater than green tea infusion (81.6 ± 0.9% vs. 57.5 ± 1.8%, P  < 0.001). The mean quercetin and kaempferol contents of roasted noni leaf infusion, as prepared by the consumer, were 0.24 ± 0.01 and 0.14 ± 0.01 μg mL⁻¹, respectively. Tannic acid content was 10 ± 1 μg mL⁻¹. The infusion was non-mutagenic in the reverse mutation test in Salmonella typhimurium and did not induce primary DNA damage in E. coli PQ37. Further, no significant primary DNA damage was induced by 5,15-dimethylmorindol, which was the only detectable anthraquinone in noni leaves. The infusion was not cytotoxic in the 24 h brine shrimp toxicity test (LC₅₀ > 1 mg mL⁻¹), nor was there any evidence of acute oral toxicity from the freeze–dried infusion in Sprague–Dawley rats (LD₅₀ > 2000 mg kg⁻¹ b.w.).     

EFFECT OF GAMMA IRRADIATION ON AFLATOXIN B1 PRODUCTION BY ASPERGILLUS FLAVUS IN GROUND BEEF STORED AT 5C

The effect of γ-irradiation for controlling the production of aflatoxin B₁ by Aspergillus flavus in ground beef stored at 5C for 2 weeks was investigated. Aspergillus, Penicillium, Cladosporium, Mucor, Scopulariopsis, Candida and Rhodotorula were the most common fungal genera contaminating ground beef. A. flavus and A. niger were the most common Aspergillus spp. Aspergillus flavus isolates were able to produce aflatoxin B₁ in ground beef. Only 3 (20%) samples of ground beef were contaminated with aflatoxin B₁ (25–45 μg/Kg). Gamma irradiation dose levels resulted in an immediate reduction in the total numbers of A. flavus. No growth or aflatoxin B₁ production occurred at 1.50 kGy during storage.     

Anti-elastase and anti-hyaluronidase of phenolic substance from Areca catechu as a new anti-ageing agent

Synopsis We have previously screened 150 medicinal plants for the inhibition of elastase and found significant inhibitory effects of the extracts of Areca catechu L. on the ageing and inflammation of skin tissues . To isolate and identify the compounds having biological activity, they were further purified by each fraction of solvents, silica gel column chromatography, preparative TLC and reversed-phase HPLC. The peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as a phenolic substance by using various colorimetric methods, UV and IR. IC₅₀ values of this phenolic substance were 26.9 μg mL⁻¹ for porcine pancreatic elastase (PPE) and 60.8 μg mL⁻¹ for human neutrophil elastase (HNE). This phenolic substance showed more potent activity than that of reference compounds, oleanolic acid (76.5 μg mL⁻¹ for PPE, 219.2 μg mL⁻¹ for HNE) and ursolic acid (31.0 μg mL⁻¹ for PPE, 118.6 μg mL⁻¹ for HNE). According to the Lineweaver–Burk plots, the inhibition against both PPE and HNE by this phenolic substance was competitive inhibition with the substrate. The phenolic substance from A. catechu effectively inhibited hyaluronidase activity (IC₅₀ : 210 μg mL⁻¹ ).
These results suggest that the phenolic substance purified from A. catechu has an anti-ageing effect by protecting connective tissue proteins.     

INFLUENCE OF MOISTURE CONTENT AND STORAGE TEMPERATURE ON THE PRODUCTION OF AFLATOXIN BY ASPERGILLUS FLAVUS EA-81 IN MAIZE AFTER EXPOSURE TO GAMMA RADIATION

The effect of gamma radiation on aflatoxin production by Aspergillus flavus EA-81 in maize with different initial moisture levels was determined over a 15-day period. The viability of A. flavus on maize decreased over time with increasing moisture contents and storage at 8C. After 45 days at 28C, levels of viable conidiospores of A. flavus increased from 4.5 × 10⁷ to about 3.0 × 10⁸ per gram of maize. Levels of aflatoxin B₁ produced by A. flavus were 10 μg kg^-1 in the maize stored at 8C after 45 days. Production of aflatoxin was highest at 40% moisture and 28C. Irradiation of 1.0 or 2.0 kGy greatly reduced the level of mold growth relative to unirradiated controls. A dose of 4.0 kGy eliminated all viable fungi. Aflatoxin B₁ production decreased with increased levels of irradiation and was negligible at 4.0 kGy. When maize was inoculated after irradiation and stored, the spore counts and aflatoxin levels were higher than in unirradiated and inoculated controls after 30 days. Apparently, the natural competitive microflora prevented growth and thus limited higher concentrations of aflatoxin in maize.     

AFLATOXIN PRODUCTION ON TWO MUSHROOM SUBSTRATES

Two fungi, Boletus edulis and Agaricus bisporus, were tested as substrates for two known aflatoxigenic fungi, Aspergillus flavus ATCC 15548 and A. parasiticus NRRL 2999. Both autoclaved substrates supported mycelial growth, sporulation, and aflatoxin production; however, the B. edulis substrate allowed more rapid mold growth and greater toxin production than did the A. bisporus substrate under laboratory conditions. Both aflatoxins B₁ and AFG₁ were produced with AFG₁ being the predominant toxin. Aflatoxins B₂ and AFG₂ were not detected. Although toxin was produced at low levels, the highest mean being 0.55 μg/g substrate for AFB₁ and AFG₁, both mushrooms apparently contained minimal nutrients for toxigenic mold growth and failed to cause antimycotic or antiaflatoxigenic responses. Routinely used aflatoxin extraction and analytical procedures appear applicable for such testing of mushrooms.     

Natural preservative ingredient from marine alga Ulva lactuca L.

The contents of total chlorophyll (T-Chl), carotenoids and phenolics compounds were quantified in the biomasses of Ulva lactuca grown either in normal or artificial sea water under indoor conditions. The antioxidant and antibacterial activities of U. lactuca crude organic extracts ( Ulva- COEs) were determined. Thirty-four compounds in Ulva- COEs were characterised by thin layer chromatography and high-performance liquid chromatography. The major compounds were chlorophyll a (Chl a ) (15.60–30.90%) and b (Chl b ) (12.20–14.89%) , 9-cis β-carotene (13.12–14.47%), α-carotene (11.44–11.47%) and all-trans β-carotene (6.16–29.70%, of total carotenoids).The Ulva- COEs exhibited remarkable antioxidant activity, with an IC₅₀ (concentration which causes a 50% of DPPH radical scavenging activity) values ranged from 16.5 and 18.7 μg mL⁻¹, which could be compared with the synthetic antioxidants: α-tocopherol (14.4 μg mL⁻¹), butylated hydroxyanisol (13.1 μg mL⁻¹) and butylated hydroxyltoluene (13.1 μg mL⁻¹). Also, Ulva- COEs exhibited great potential antibacterial activities against six bacterial strains, with minimal inhibitory concentration values ranged from 0.40 to 0.35 mg mL⁻¹.     

Optimising the free radical scavenging activity of shrimp protein hydrolysate produced with alcalase using response surface methodology

Response surface methodology (RSM) was used to optimise the hydrolysis parameters of Acetes chinensis by Alcalase 2.4L in order to obtain a hydrolysate with potent radical scavenging activity. The parameters were temperature, pH and enzyme concentration/substrate concentration (E/S) ratio with degree of hydrolysis (DH) being the response. The results showed that the optimum condition was: temperature at 57 °C, pH at 8.0, E/S ratio at 2.6AU 100 g⁻¹ shrimp, hydrolysis time 3 h. The DH was 26.32%, the hydroxyl radical scavenging activity was up to 88.12% and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was 35.61%. The gel column filtration chromatography by a Sephadex G-25 column yielded five fractions. The molecular weight of the most potent free radical scavenging activity fraction ranged from 915 to 207 Da, its IC₅₀ for hydroxyl radical was 0.03 mg mL⁻¹, and IC₅₀ for DPPH radical was 8.86 mg mL⁻¹.     

Storage stability of a stimulant coconut water–acerola fruit juice beverage

The objective of this work was to study the stability of a beverage formulated with acerola fruit juice and green coconut water with added caffeine. The beverage was prepared with 25% acerola pulp, 75% green coconut water and sugar up to 12°Brix, and caffeine (125 mg L⁻¹), heat processed at 90 °C for 30 s and packed in 250-mL glass bottles. Chemical, physicochemical, microbiological and sensory analyses of the beverage were performed just after processing and during 6 months of storage at room temperature (27 °C). The vitamin C content decreased significantly throughout storage, from 399.5 to 189.6 mg 100 mL⁻¹, although it has remained relatively high. The anthocyanins initially present (0.025 mg 100 mL⁻¹) were completely lost during the storage at a mean rate of 4 μg 100 mL⁻¹ month⁻¹. The product was microbiologically stable during storage. Colour changes were also observed with absorbance at 420 nm, with average values ranging from 0.19 to 0.24. However, according to the sensory analyses the product was acceptable during the 6 months of storage, presenting sensory scores (colour, taste and global acceptance) from 6.5 to 5.5, which suggests its potential for market.     

An evaluation of extracts of five traditional medicinal plants from Iran on the inhibition of mushroom tyrosinase activity and scavenging of free radicals

This study aimed to evaluate the free radical scavenging and inhibition properties of five medicinal plants, including Quercus infectoria Olive., Terminalia chebula Retz. , Lavendula stoechas L., Mentha longifolia L., Rheum palmatum L., toward the activity of mushroom tyrosinase using l -tyrosine and l -3,4-dihydroxyphenylalanine ( l -DOPA) as the substrate. The methanol extracts of Q. infectoria and T. chebula showed strong radical scavenging effect in 2,2'-dipheny l -1-picrylhydrazyl (DPPH) assay (IC₅₀ = 15.3 and 82.2 μg mL⁻¹ respectively). These plants also showed inhibitory effects against the activity of mushroom tyrosinase in hydroxylation of l -tyrosine (85.9% and 82.2% inhibition, respectively). These two plants also inhibited the oxidation of l -DOPA similar to kojic acid as positive control (IC₅₀ = 102.8 and 192.6 μg mL⁻¹ respectively). In general Q. infectoria and T. chebula significantly inhibited tyrosinase activity and DPPH radical. Both activities were concentration-dependant but not in linear manner. It is needed to study the cytotoxicity of these plant extracts in pigment cell culture before further evaluation and moving to in vivo conditions.     

INFLUENCE OF OTHER FUNGI ON AFLATOXIN PRODUCTION BY ASPERGILLUS FLAVUS IN MAIZE KERNELS

Experiments were designed to determine whether certain nontoxigenic fungi commonly isolated from maize kernels can affect aflatoxin B₁ development when inoculated with A. flavus onto individual unsterilized, and autoclaved maize kernels . Trichoderma viride and Aspergillus niger were found to be strongly antagonist inhibiting the growth of A. flavus by 87 and 66% respectively, whereas Aspergillus versicolor, Fusarium moniliforme, Paecilomyces variotii and Emericella quadrillineata inhibited the growth of A. flavus by less than 51%. Less aflatoxin B₁ was detected when A. flavus was paired with A. niger or T. viride than with the other test fungi. When A. niger or T. viride was introduced onto the kernels 72 h before inoculation with A. flavus, no aflatoxin B₁ was detected in unsterilized kernels and the levels of aflatoxin B₁ were greatly reduced from 700 ppb to 160 and 140 ppb in autoclaved kernels, respectively. When inoculation of A. flavus followed 72 h of incubation of either A. niger and T. viride, no aflatoxin B₁ was detected. However, when both A. niger and T. viride were introduced 72 h after inoculation with A. flavus, the levels of aflatoxin B₁ were reduced to 385 and 560 ppb, respectively in unsterilized and autoclaved maize kernels . Trichoderma viride and Aspergillus niger may be useful in biological control of aflatoxin contamination of maize kernels .; Accepted for Publication June 11, 1997     

The impact of both the season of collection and drying on the volatile constituents of Origanum vulgare L. ssp. hirtum grown wild in Croatia

Samples of Origanum vulgare ssp. hirtum were collected from the same geographic area in the south of Croatia at different seasons of growth. The maximum fluctuations found for the main components from fresh plant material were: thymol [149.2–1124.4 mg (100 g)^–1], carvacrol [51.6–564.3 mg (100 g)^–1], p-cymene [20.2–220.9 mg (100 g)^–1] and γ-terpinene [50.1–217.5 mg (100 g)^–1]. The oregano that was analysed belonged to a thymol/carvacrol chemotype. The season of collecting affected the qualitative and quantitative composition of the essential oil. The most impressive difference was the increase of p-cymene content in August. After the drying of the plant material, all samples showed a minor decrease in essential oil yields when compared with fresh plants. Drying, at room temperature, had no effect on the qualitative composition of oregano oil. Because of the variability of essential oil compositions from seasonally collected fresh and dried oregano, it would be important to check the quantity and quality of such components before usage.     

A modified colorimetric method for the estimation of β-cyclodextrin using phenolphthalein

The binding equilibrium between β-cyclodextrin and phenolphthalein has been used to develop a method for the estimation of β-cyclodextrin in solution. From logarithmic plots of amounts of β-cyclodextrin against absorbance at 554 nm, a relation log X  = (log A  − log Y )/ B was found, which gave an estimate of β-cyclodextrin in the concentration range 0.0045 mg mL⁻¹ (3.96 μm) to 4.7 mg mL⁻¹ (4.14 mm), where X =  intercept and B  = slope. This method was found to be highly reproducible and reliable.     

Natural maize phenolic acids for control of aflatoxigenic fungi on maize

ABSTRACT:  Natural phytochemicals may be an alternative to synthetic chemicals for controlling fungal growth and mycotoxin production in stored maize. A key to progress in this field is to select the best natural maize phytochemicals to be applied in a storage maize ecosystem. This research was undertaken to evaluate the effects of the natural phytochemicals trans-cinnamic acid (CA) and ferulic acid (FA) alone at concentrations of 20 to 30 mM and in 5 combinations on Aspergillus flavus Link and A. parasiticus Speare populations and aflatoxin B₁ production. Studies on Aspergillus population and aflatoxin B₁ production were carried out in maize grain in relation to a water activity a_w of 0.99, 0.97, 0.95, and 0.93. CA and FA at concentrations of 25 to 30 mM, respectively, and CA-FA mixture T9 (25 + 30 mM) were the treatments most effective at inhibiting A. flavus and A. parasiticus population at all a_w assayed after 11 d of incubation. At all a_w values, the mixture CA-FA T9 (25 + 30 mM) completely inhibited (100%) aflatoxin B₁ production by both strains at a_w= 0.99, 0.97, 0.95, and 0.93. Decreased aflatoxin B₁ levels in comparison with the control were observed with mixtures CA-FA T6 (10 + 25 mM), T7 (20 + 20 mM), and T8 (20 + 30 mM) of both strains in the majority of a_w assayed. The data show that CA and FA could be considered as effective fungitoxicants for A. flavus and A. parasiticus in maize in the a_w range 0.99 to 0.93. The information obtained shows promise for controlling aflatoxigenic fungi in stored maize.     

Studying the anti-tyrosinase effect of Arbutus andrachne L. extracts

Arbutus andrachne L. is widely distributed in Jordan. Tyrosinase is the key enzyme in the biosynthesis of melanin. This preliminary study was carried out to assess the possible anti-tyrosinase activity of A. andrachne extracts. Arbutin, hydroquinone and kojic acid were selected as inhibitor standards. Five different extracts (chloroform, butanol, ethanol, methanol and water) were prepared from A. andrachne stems and their activities were compared with the selected tyrosinase inhibitors. IC₅₀ was measured for both, standard and plant extracts. Among the different extracts, the methanolic extract exhibited the highest anttyrosinase activity with an IC₅₀ value (1 mg mL⁻¹). Furthermore, 9 mg A. andrachne methanolic extract showed 97.49% inhibition of tyrosinase activity. Arbutin, hydroquinone, β-sitosterol and ursolic acid were identified in the different extracts of A. andrachne by thin layer chromatography (TLC) and isolated by preparative TLC from the methanolic and chloroform stem extracts, respectively.     

Aflatoxin levels in dried figs, nuts and paprika for export from Turkey

Three hundred and thirteen of 2643 dried fig, two of eighty hazelnut, sixteen of twenty-eight pistachio, five of ten peanut and nineteen of twenty-three paprika samples for export from Turkey were contaminated with total aflatoxins in the range of 0.2–162.76, 5.46–6.55, 2.31–63.11, 0.75–26.36 and 1.79–6.55 μg kg⁻¹, respectively. Samples were collected from January to August 2007 and tested for aflatoxins (B₁, B₂, G₁ and G₂) by immunoaffinity column extraction using RP-HPLC. Fifty-six of the 313 dried fig, all of the contaminated hazelnut and pistachio, two of the sixteen peanut and three of the nineteen paprika samples exceeded the regulatory limits of the European Union. The ratio of the different types of aflatoxin present in each sample exhibited great variability. For example, of 313 contaminated fig samples, 159 contained only aflatoxin B₁, eighty-five contained B₁ (49.7%) + G₁ (50.3%), twenty-two contained only G₁, twenty contained B₁ (89.4%) + B₂ (10.6%), thirteen contained B₁ (73.7%) + B₂ (10.8%) + G₁ (15.5%) and fourteen contained all four types, B₁ (26%) + B₂ (2.5%) + G₁ (66.5%) + G₂ (5%).     

DPPH free-radical scavenging ability, total phenolic content, and chemical composition analysis of forty-five kinds of essential oils

Forty-five kinds of commonly used essential oils were employed to investigate the DPPH (1,1-diphenyl2-picrylhydrazyl) radical scavenging ability and total phenolic content of major chemical compositions. The free-radical scavenging ability and total phenolic content of cinnamon leaf and clove bud essential oils are the best among these essential oils. One-half milliliter of cinnamon leaf and clove bud essential oils (10 mg mL EtOH) are shown to be 96.74% and 96.12% of the DPPH (2.5ml, 1.52 × 10^-4 M) free-radical scavenging ability, respectively. Their EC50 (effective concentrations) are 53 and 36 (μg mL^-1). One milligram per milliliter of cinnamon leaf, clove bud, and thyme red essential oils were shown to be 420, 480, and 270 (mg g^-1 of GAE) of total phenolic content, respectively. Eugenol in cinnamon leaf and clove bud essential oils (82.87% and 82.32%, respectively) were analyzed by GC-MS. It is clear that the amounts of the phenol compounds in essential oils and the DPPH free-radical scavenging ability are in direct proportion.