Effect of water activity on stress tolerance and biocontrol activity in antagonistic yeast Rhodosporidium paludigenum

The aims of this study were to optimize the salt-adaptation conditions of the marine antagonistic yeast Rhodosporidium paludigenum and investigate the biocontrol activity of salt-induced cell suspensions of R. paludigenum on postharvest pathogens in fruits. Low water activity (a_w = 0.98, 0.97, 0.96, and 0.95) inhibited the growth of R. paludigenum in nutrient yeast dextrose broth, but the yeast grew better in the medium modified with NaCl solute than other nonionic solutes. R. paludigenum grown in 6.6% NaCl-modified medium had higher viabilities (92.1%) at low water activity (a_w = 0.95) than control (81.1%) after 48 h incubation. The salt-adapted R. paludigenum also showed better viability than the un-adapted cells after being frozen, which may be related to the accumulation of intracellular trehalose. Moreover, the best biocontrol inhibition in pears and Chinese winter jujubes was obtained when R. paludigenum was grown in NaCl-modified medium. Therefore, this study implies that improving physiological inducement methods may be a promising strategy for accelerating commercialization of biocontrol agents.     

Flavonoids of Zizyphus jujuba L. and Zizyphus spina-christi (L.) Willd (Rhamnaceae) fruits

The ripe edible fruits of jujube, Zizyphus jujuba Miller (syn. Z. sativa Gaertner, Z. vulgaris Lam.) and Christ’s thorn jujube Zizyphus spina-christi (L.) Willd (Rhamnaceae family) were phytochemically investigated, comparing their quali-quantitative flavonoids profile. Twelve compounds from both methanol extracts have been recognized as quercetin, kaempferol, and phloretin derivatives by means of HPLC/ESI-MS analyses. Six major compounds have been purified by Sephadex LH-20 column chromatography followed by HPLC and were characterized using NMR spectroscopy. One C-glycoside, 3′,5′-di-C-β-d-glucosylphloretin, was detected in Z. spina-christi. The quantitative analysis of all compounds was also reported showing a higher content of flavonoids in Z. jujuba.     

Potential roles of essential oil and organic extracts of Zizyphus jujuba in inhibiting food-borne pathogens

This study was undertaken to examine the chemical compositions of essential oil and tested the efficacy of oil and organic extracts from seeds of Zizyphus jujuba against food-borne pathogens. The chemical compositions of the oil was analysed by Null. Twenty three compounds representing 91.59% of the total oil were identified. The oil (5 μl of 1:5 (v/v) dilution of oil with methanol) and extracts of hexane, chloroform, ethyl acetate and methanol (300 μg/disc) of Z. jujuba displayed a remarkable antibacterial activity against Staphylococcus aureus (ATCC 6538 and KCTC 1916), Listeria monocytogenes ATCC 19166, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa KCTC 2004, Salmonella typhimurium KCTC 2515 and Escherichia coli ATCC 8739. The scanning electron microscopic studies also demonstrated the effect of essential oil on the morphology of Staph. aureus ATCC 6538 at the MIC value, along with the potential effect on cell viabilities of the tested bacteria.     

The effects of the combination of Pichia membranefaciens and BTH on controlling of blue mould decay caused by Penicillium expansum in peach fruit

The feasibility of 0.2 g l⁻¹ benzo-thiadiazole-7-carbothioic acid S-methyl ester (BTH) to improve the efficacy of Pichia membranefaciens in controlling postharvest blue mould decay in peach fruit was investigated. Our results showed that biocontrol activity of P. membranefaciens against blue mould caused by Penicillium expansum in peach fruit could be enhanced by addition of 0.2 g l⁻¹ BTH. The combination of P. membranefaciens and BTH resulted in a more effective control of blue mould than individual treatment of P. membranefaciens or BTH alone. The combined treatment had a synergistic effect on the induction of superoxide dismutase, catalase, ascorbate peroxidase, chitinase and β-1,3-glucanase activities, which induced stronger disease resistance in fruit than BTH or yeast alone, and resulted in a lower lesion diameter and disease incidence of blue mould decay in peaches. Furthermore, the combined treatment did not impair the quality parameters including fruit firmness and contents of total soluble solids, titratable acidity and vitamin C of peach fruit after 6 days of storage at 20 °C. These results suggested that the use of BTH may be an effective method to improve the biological activity of P. membranefaciens.     

Salicylic acid enhances biocontrol efficacy of Rhodotorula glutinis against postharvest Rhizopus rot of strawberries and the possible mechanisms involved

The potential of using Rhodotorula glutinis alone or in combination with salicylic acid (SA) for the control of postharvest Rhizopus rot of strawberries, and their effects on enzyme activities of fruits were investigated. The combination of R. glutinis (1 × 10⁸ CFU ml⁻¹) with SA (100 μg ml⁻¹) resulted in a significant reduction in the disease incidence and lesion diameter of Rhizopus rot on the strawberry fruits at 20 °C and 4 °C, and more so than with SA or yeast alone. SA at the concentration of 100–1000 μg ml⁻¹ significantly inhibited spore germination of Rhizopus stolonifer. About 100 μg ml⁻¹ of SA did not inhibit the growth of the antagonistic yeast, and could significantly increase the population growth of R. glutinis in strawberry wounds at 20 °C. SA, combined with R. glutinis, increased the activity of strawberry host defence enzymes (POD) and cell wall lytic enzymes (β-1,3-glucanase).     

Effects of nitric oxide fumigation on phenolic metabolism of postharvest Chinese winter jujube (Zizyphus jujuba Mill. cv. Dongzao) in relation to fruit quality

The effect of nitric oxide fumigation on phenolic metabolism of harvested Chinese winter jujube in relation to the fruit quality was investigated. The fruits were fumigated for 3 h with NO (0, 10, 20 and 30 μl/l) then stored at 22 °C and RH 95%. Changes in color, phenol and anthocyanin levels in pericarp, activities of the related enzymes, and total soluble solids and vitamin C from mesocarp were measured. The results showed that treatment with 20 μl/l NO significantly slowed the increase in red index, inhibited changes of polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) activities, maintained a low total anthocyanin content and a high total phenol content, and delayed the increase of soluble solids and decrease of vitamin C. Treatment with NO solution at less than 1 μmol/l exhibited inhibitory effects on in vitro PPO and PAL activities in a dose-dependent manner.     

Methyl jasmonate enhances biocontrol efficacy of Rhodotorula glutinis to postharvest blue mold decay of pears

The effect of Rhodotorula glutinis treatment alone or in combination with methyl jasmonate (MeJA) in controlling blue mold decay, the natural fungal decay of pears and the postharvest quality parameters including fruit firmness, total soluble solids, titratable acidity, and ascorbic acid were investigated. The combination of methyl jasmonate (200 μM) and R. glutinis (1 × 10⁸ CFU/ml) was a more effective approach to reduce the disease incidence and lesion diameter of blue mold decay of pears than the application of MeJA or R. glutinis alone after incubation for 7 d at 20 °C. The natural fungal decay of pears treated with the application of R. glutinis combined with MeJA resulted in reduced average decay incidence of 10.42% or 4.16%, respectively, compared with 27.17% or 20.83% in the control fruits following storage at 20 °C for 15 d or 4 °C for 60 d followed by 20 °C for 15 d. The combined treatment did not impair quality parameters of fruits under both conditions.     

Soluble phenols and antioxidant activity in mamey sapote (Pouteria sapota) fruits in postharvest

Mamey sapote (Pouteria sapota (Jacq.) H. E. Moore & Stearn) is a tropical species from the Sapotaceae family native to Mexico and Central America. Its tree produces an edible climacteric fruit, whose weight can range from 250 to 900 g. The flesh is soft, comprises about 78% of the fruit, and has high sugars to acidity ratio, which gives it a sweet taste when ripe. Because of current public interest to consume products that promote health, the aim of the work was to characterize soluble phenolic content and antioxidant activity in mamey sapote fruits from Chiapas, Mexico, at different maturity stages. Storage at 12 (± 1) °C was conducted for 18 d. When fruits ripened firmness flesh varied, in average, from 41.8 to 3.0 N, soluble solids content from 17.7 to 28.1 ºBrix, hue angle from 56.4 to 46.3º, and lightness from 67.3 to 42.0. Phenolic content was affected by ripening since average values of 2563 and 234 ??g/g were found in unripe and consumption maturities, respectively. In a soluble phenolics extract from flesh, gallic acid (GA), gallocatechin-3-gallate (G3G), epicatechin (ECT), and catechin (C) were found, being the latter the most abundant compound, which increased with ripening from 9.9 to 113.1 ??g/g, while GA, G3G, and ECT had average values of 4.7, 11.9, and 5.8 ??g/g, respectively, without significant variation. Antioxidant activity, expressed through the IC₅₀ parameter, remained practically unchanged and showed an average value of 12.9 ??g/mL. Based on phenolic composition the mamey sapote fruit may constitute a good source of antioxidant compounds.     

Profiling of in vitro neurobiological effects and phenolic acids of selected endemic Salvia species

The ethyl acetate and methanol extracts from 16 Salvia L. species were screened for their inhibitory activity against acetylcholinesterase, butyrylcholinesterase, lipoxygenase, and tyrosinase; the enzymes linked to neurodegeneration. Their antioxidant activity was also tested using DPPH radical scavenging, metal-chelation, and ferric-reducing antioxidant power (FRAP) assays. Total flavonoid content of the extracts was determined by AlCl₃ reagent, while HPLC technique was applied for analysis of various phenolic acids in the extracts. The extracts exerted weak cholinesterase and tyrosinase inhibition, and remarkable inhibition against lipoxygenase (13.07 ± 2.73-74.21 ± 5.61%) at 100 μg ml⁻¹. The methanol extracts showed higher antioxidant activity in DPPH radical scavenging and FRAP assays. The extracts were analyzed for their gallic, protocateuchic, p-hydroxy-benzoic, vanillic, caffeic, chlorogenic, syringic, o- and p-coumaric, ferulic, rosmarinic, and tr-cinnamic acid contents and the methanol extract of Salvia ekimiana (153.50 mg 100 g⁻¹) was revealed to be the richest in terms of rosmarinic acid.     

Enzymatic properties of transglutaminase produced by a new strain of Bacillus circulans BL32 and its action over food proteins

The aim of this study was to physicochemically characterize transglutaminase (TGase) from Bacillus circulans BL32, a strain recently isolated from the Amazon basin region, for its application in food systems. The effects of pH and temperature on the enzyme activity were determined by Central Composite Rotatable Design (CCRD), with maximal TGase activities obtained for pH between 5.7 and 8.7 and temperatures of 25-45 °C. This microbial TGase showed to be remarkably stable: over 90% of its activity was retained after 120 min of incubation at 50 °C. The Ca²⁺ and Mg²⁺ cations enhanced enzyme activity and its thermal stability when in concentrations of up to 2 and 1 mol L⁻¹, respectively. Casein, isolated soy protein, and hydrolysed animal protein were treated with this TGase. The decrease in the amount of free amino groups, especially for casein, showed the cross-linking of protein catalysed by this enzyme, while the emulsifying properties of these proteins were improved with treatment. These results suggest that this microbial TGase has a good potential to be used in food and other industrial applications.     

The potential of Cycloclasticus and Altererythrobacter strains for use in bioremediation of petroleum-aromatic-contaminated tropical marine environments

Cycloclasticus sp. A5, which has been suggested to be a major degrader of petroleum aromatics spilled in temperate seas, showed higher degrading activities for petroleum aromatics, at both 25 °C and tropical sea temperature 30 °C, than the novel aromatic-degrading isolates, related to Altererythrobacter epoxidivorans (97.5% similarity in the almost full-length 16S rRNA gene sequence) and Rhodovulum iodosum (96.3% similarity), obtained after enrichment on crude oil in a continuous supply of Indonesian seawater. Cycloclasticus A5 degraded petroleum aromatics at a similar rate or faster at 30 °C as compared to 25 °C, but its growth on acetate was severely inhibited at 30 °C. These results suggest that, although their abundance would be low in tropical seas not contaminated with aromatics, the Cycloclasticus strains could be major degraders of petroleum aromatics spilled in tropical seas. The 16S rRNA gene of the Cycloclasticus strains has been identified from Indonesian seawater, and the gene fragments showed 96.7−96.8% similarities to that of Cycloclasticus A5. Introducing Cycloclasticus A5 may be an ecologically advantageous bioremediation strategy for petroleum-aromatic-contaminated tropical seas because strain A5 would disappear at 30 °C after complete consumption of the aromatics. Altererythrobacter and Rhodovulum-related isolates grew well on pyruvate in 10% strength marine broth at 30 °C whereas Cycloclasticus A5 did not grow well on acetate in the broth at 30 °C. These growth results, along with its petroleum-aromatic-degrading activity, suggest that the Altererythrobacter isolate could be an important petroleum-aromatic degrader in and around nutrient-rich tropical marine environments.     

Antibacterial activity of the extracts from the fruit rinds of Garcinia cowa and Garcinia pedunculata against food borne pathogens and spoilage bacteria

The crude hexane and chloroform extracts from the fruit rinds of Garcinia cowa and Garcinia pedunculata were studied for their antibacterial activity against some foodborne pathogens and spoilage bacteria such as Bacillus cereus, Bacillus coagulans, Bacillus subtilis, Staphylococcus aureus and Escherichia coli. The minimum inhibitory concentrations (MICs) of the extracts determined by the agar dilution method were ranging from 15 to 500 μg/ml and 300 to 1250 μg/ml for G. cowa and G. pedunculata, respectively. However, the hexane and chloroform extracts from the fruit rinds of G. cowa exhibited marked inhibitory effect against all the test organisms and were more effective than that of G. pedunculata extracts. The antibacterial activity of all the extracts was more pronounced against the tested Gram-positive bacteria than the tested Gram-negative bacterium. Furthermore, this study is the first report on the in vitro antibacterial activity of extracts from the fruit rinds of G. cowa and G. pedunculata.     

Glucosinolate content and myrosinase activity evolution in three maca (Lepidium meyenii Walp.) ecotypes during preharvest,harvest and postharvest drying

Glucosinolate profiles, glucosinolate contents and myrosinase activity were evaluated in yellow, red and black hypocotyls of maca during pre-harvest, at harvest and during post-harvest drying. At harvest, six glucosinolates (GLs) were identified: 5-methylsulfinylpentyl, 4-hydroxybenzyl, benzyl, 3-methoxybenzyl, 4-hydroxy-3-indolylmethyl and 4-methoxy-3-indolylmethyl, of which benzyl glucosinolate was the most abundant in the three ecotypes, representing 80% of the total GLs. A significant increase in GLs was observed for the three ecotypes during the 90 days before harvest and during the 15–30 days of post-harvest drying. This was followed by an important decrease of GLs during the 30–45 day period, which was attributed to cell breakdown, due to fluctuations in temperatures during the drying process, and was correlated with a high myrosinase action. During the last period of post-harvest drying, GLs were much lower and correlated to lower myrosinase activity and lower maca hypocotyl humidity. A combination of artisanal and other processing techniques should be utilised, in order to best preserve maca glucosinolates.     

Phytochemicals and antioxidant activity of different parts of bambangan (Mangifera pajang) and tarap (Artocarpus odoratissimus)

Mangifera pajang (family: Anacardiaceae; local name: bambangan) and Artocarpus odoratissimus (familiy: Moraceae; local name: tarap) are popular edible fruits in Sabah, Malaysia. The flesh, kernel and peel from M. pajang; seed and flesh from A. odoratissimus were analysed for total antioxidant activity, total polyphenol, total flavonoid and total anthocyanins contents. M. pajang kernel extract displayed the highest free radical scavenging and ferric reducing activities. Total phenolic content of the samples were in the range of 5.96–103.3 mg gallic acid equivalent/g. M. pajang kernel and M. pajang flesh contained the highest and lowest total flavonoid content with the values of 10.98 and 0.07 mg rutin equivalent/g, respectively. The antioxidant activities of extracts were significantly correlated with the total phenolic and flavonoid content (but not the anthocyanins content). The phytochemicals and antioxidant properties of M. pajang and A. odoratissimus, especially their by-products (kernel/seed), indicate that they may impart health benefits when consumed and should be regarded as a valuable source of antioxidant-rich nutraceuticals.     

Inhibitory effect of sour orange (Citrus aurantium) juice on Salmonella Typhimurium and Listeria monocytogenes

The sour orange (Citrus aurantium) juice is commonly used as flavoring and acidifying agent for vegetable salads and appetizers in Turkey. It was aimed to determine the survival and growth pattern of Salmonella Typhimurium and Listeria monocytogenes in sour orange juice. Different concentrations of neutralized and un-neutralized juice samples were inoculated with each of the test microorganisms (∼6 log CFU/mL) separately and then incubated at 4 °C and 37 °C for seven days. It was detected both of the test microorganisms could survive and even grow in neutralized juice samples at 37 °C for two days. However, none of them could survive at the end of seventh day of incubation at 37 °C. Low incubation temperature (+4 °C) increased the survival of the tested microorganisms. Also, it was detected that L. monocytogenes were less resistant to the variable conditions than S. Typhimurium. It was concluded that the antimicrobial effect of sour orange juice mainly depends on the low pH value of the product. However, incubation time and temperature are also effective on the survival of the tested pathogens.     

The in vitro and in vivo effects of yerba mate (Ilex paraguariensis) extract on adipogenesis

The aim of this study was to evaluate the effects of yerba mate extract and its principal bioactive compounds on adipogenesis. The anti-adipogenic effects of yerba mate, chlorogenic acid, quercetin and rutin were evaluated in 3T3-L1 cells using a PCR array. The results obtained in vitro were validated in vivo in a high-fat diet-induced model of obesity. The in vitro and in vivo results demonstrated that yerba mate extract down-regulated the expression of genes that regulate adipogenesis, such as Creb-1and C/EBPα, and the extract up-regulated the expression of genes related to the inhibition of adipogenesis, including Dlk1, Gata2, Gata3, Klf2, Lrp5, Pparγ₂, Sfrp1, Tcf7l2, Wnt10b, and Wnt3a. In summary, it was demonstrated that yerba mate and its bioactive compounds regulate the expression of genes related to in vitro adipogenesis. Furthermore, yerba mate might regulate adipogenesis through the Wnt pathway.     

Influences of preharvest spraying Cryptococcus laurentii combined with postharvest chitosan coating on postharvest diseases and quality of table grapes in storage

The effects of preharvest spray with Cryptococcus laurentii combined with chitosan coating after harvest on decay and quality of table grapes during storage periods were evaluated in the present study. Preharvest spray with C. laurentii (PreA) significantly decreased decay index (DI), and postharvest chitosan coating (PCC) enhanced the effectiveness of the pre-harvest spray when fruits were stored at 0 °C. PreA combination with PCC increased the activities of polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) of fruit in storage. PreA + PCC treatment was effective in reducing weight loss of fruits by 85% at 17 d storage and 38% at 42 d storage as compared to PreA alone at the same stage. In addition, PreA enhanced the ratio of soluble solids content (SSC) to titratable acid (TA) by 12% at harvest time, 7% at 17 d storage and 25% at 42 d storage, mainly by increasing SSC and decreasing TA in fruit stored at 0 °C. These results suggested that integration of preharvest spray with C. laurentii and postharvest chitosan coating treatment may be a promising management strategy for decay control and quality maintenance of table grapes.     

Studies on use of Enteromorpha in snack food

Seaweeds, also known as sea vegetables, are of nutritional interest, as they are rich in vitamins, minerals and dietary fiber. Enteromorpha compressa (Linnaeus), green seaweed (chlorophyta), which is a rich source of iron and dietary fibre was used as an ingredient in the preparation of Pakoda, a common traditional snack food in India. Pakoda samples showed increases in ash, protein and total dietary fibre contents with increase in Enteromorpha level, accompanied by a nearly fivefold increase in iron content (26.4–126 mg/100 g) and fourfold increase in calcium content (30.1–124 mg/100 g). Bioavailability of iron in Enteromorpha, and Pakoda containing 7.5% of Enteromorpha, did not show any difference (55–56%) at pH 7.5 (intestinal condition). But, at pH 1.35 (gastric condition) the bioavailability of iron in Pakoda containing Enteromorpha was found to be slightly higher (27.1%) than that in Enteromorpha. Reducing power (155–222 μg/g) increased as the Enteromorpha level increased from 0% to 15%. But the addition of Enteromorpha was found to decrease free radical-scavenging activity and the total phenol content. Pakoda containing up to 7.5% of Enteromorpha was found to have a sensory quality comparable with that of Pakoda without Enteromorpha.     

A laboratory assessment of the potential of different strains of the entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) to control Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) in stored cowpea

Twelve indigenous and exotic isolates of Beauveria bassiana and Metarhizium anisopliae were evaluated for their virulence and their ability to suppress populations of Callosobruchus maculatus in stored cowpea. LT₅₀ values ranged from 3.11 to 6.12 days following immersion in aqueous suspensions containing 1×10⁸ conidia ml⁻¹. Indigenous isolates that had been recovered from C. maculatus were more virulent in laboratory bioassays than exotic isolates from other insects. The two isolates with the shortest LT₅₀ values were compared in dose-response assays by immersion and by exposure to cowpea grains treated with dry conidia. In both assays B. bassiana 0362 was consistently more virulent than M. anisopliae 0351. By immersion, LC₅₀ values on day 6 post-treatment were 9.10×10⁴ and 7.10×10⁵ conidia ml⁻¹ for B. bassiana 0362 and M. anisopliae 0351, respectively. Exposure to treated grains gave LC₅₀ values of 1.15×10⁷ and 4.44×10⁷ conidia g⁻¹ grain for B. bassiana 0362 and M. anisopliae 0351, respectively. In 1 kg batches of cowpea stocked with 50 adult C. maculatus, B. bassiana 0362 at both 1×10⁷ and 1×10⁸ conidia g⁻¹ grain led to significant adult mortality and reduced F₁ emergence relative to untreated populations. At 1×10⁸ conidia g⁻¹ the effect of the fungus persisted into the F₁ generation. The net reproductive rates, R₀, measured 26 days after insects were released were 5.16 and 7.32 for the high and low doses compared to 9.52 for the untreated control.