Influence of fungicide residues on the primary fermentation of young lager beer

The effect of four sterol biosynthesis-inhibiting fungicides added to the pitching wort on the evolution of several organoleptic parameters during the primary fermentation of young lager beer was assessed. Pyrimidine (nuarimol and fenarimol) and triazole (myclobutanil and propiconazole) fungicides were individually supplied to the pitching wort to obtain a concentration of 1 mg/L. A marked influence in the fermentation rate was observed for the samples with propiconazole residues. From the fourth day onward, the fermentation prematurely ceased (stuck fermentation), and therefore, statistical significant differences were found in fermented extract, alcohol content, fermentable carbohydrates, pH, color, and total polyphenol and flavonoid contents of beer. Myclobutanil residues are only influenced in the total polyphenol and flavonoid contents, while differences in the analyzed parameters were not noticeable for the samples containing nuarimol and fenarimol residues in comparison with the blank sample.     

Residue analysis and degradation studies of fenbuconazole and myclobutanil in strawberry by chiral high-performance liquid chromatography-tandem mass spectrometry

A simple and sensitive enantioselective method for the determination of fenbuconazole and myclobutanil in strawberry was developed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Fenbuconazole and myclobutanil residues in strawberry were extracted with acetonitrile containing 1% acetic acid, and an aliquot was cleaned up with PSA (primary and secondary amine) and C(18) sorbent. The direct resolution of fenbuconazole and myclobutanil enantiomers was performed on a cellulose tris (3,5-dimethylphenylcarbamate) column using acetonitrile-0.1% formic acid solution (60:40, v/v) as the mobile phase. Quantification was achieved using matrix-matched standard calibration curves, and the limits of quantification for fenbuconazole and myclobutanil enantiomers in strawberry were both 2 μg/kg. The method was successfully utilized to investigate the probable enantioselective degradation of fenbuconazole and myclobutanil in strawberry. The results showed that the degradation of the fenbuconazole and myclobutanil enantiomers in strawberry followed pseudofirst-order kinetics (R(2) > 0.97). The results from this study revealed that the degradation of fenbuconazole in strawberry was not enantioselective, while the degradation of myclobutanil was enantioselective, and the (+)-myclobutanil showed a faster degradation than (-)-myclobutanil in strawberry, resulting in the relative enrichment of (-)-myclobutanil in residue. The results could provide a reference to fully evaluate the risks of these two fungicides.     

Fate of pesticides during beer brewing

The fates of more than 300 pesticide residues were investigated in the course of beer brewing. Ground malt artificially contaminated with pesticides was brewed via steps such as mashing, boiling, and fermentation. Analytical samples were taken from wort, spent grain, and beer produced at certain key points in the brewing process. The samples were extracted and purified with the QuEChERS (Quick Easy Cheap Effective Rugged and Safe) method and were then analyzed by LC-MS/MS using a multiresidue method. In the results, a majority of pesticides showed a reduction in the unhopped wort and were adsorbed onto the spent grain after mashing. In addition, some pesticides diminished during the boiling and fermentation. This suggests that the reduction was caused mainly by adsorption, pyrolysis, and hydrolysis. After the entire process of brewing, the risks of contaminating beer with pesticides were reduced remarkably, and only a few pesticides remained without being removed or resolved.     

Method development and fate determination of pesticide-treated hops and their subsequent usage in the production of beer

The fate of residues of seven agrochemicals (chlorfenapyr, quinoxyfen, tebuconazole, fenarimol, pyridaben, and E- and Z-dimethomorph) from the treatment on hops to the brewing of beer was studied. First, a multi-residue analytical method was developed for the determination of pesticide residues in spent hops, trub, wort, and beer. Each matrix was validated over at least two levels of fortification, for all seven compounds, in the ranges 0.05-5.0, 0.001-1.0, 0.001-0.05, and 0.0005-1.0 ppm for spent hops, trub, wort, and beer, respectively. Recoveries ranged from 73 to 136%. Second, the matrixes prepared from hops, which were treated under commercial practices with each compound, were analyzed using the method developed. The use of treated hops resulted in the carryover of 0.001 ppm of tebuconazole, 0.008 Z-dimethomorph, and 0.005 ppm of E-dimethomorph into the wort. The bulk of the remaining residues of all seven compounds was found on the spent hops. Following fermentation, all compounds were found in levels less than 0.0005 ppm in beer, except Z- (0.006 ppm) and E-dimethomorph (0.004 ppm). Third, when all seven pesticides were spiked prior to the pitching of yeast into clean wort, most of the nonpolar compounds (chlorfenapyr, quinoxyfen, and pyridaben) partitioned into the organic material (trub) which settled to the bottom, while the more polar compounds (fenarimol, tebuconazole, and E- and Z-dimethomorph) were generally distributed evenly between the beer and the trub.     

Factors affecting imazalil and thiabendazole uptake and persistence in citrus fruits following dip treatments.

The effect of concentration, temperature, and length of treatment with imazalil (IMZ) and thiabendazole (TBZ) was studied with application to citrus fruit. The amount of residues retained by fruit after "home" washing was also monitored. IMZ uptake in citrus fruit was related to treatment duration, whereas TBZ residues was not. Residues of IMZ or TBZ fungicides were significantly correlated with dip temperature (r = 0.943 for IMZ; r = 0.911 for TBZ). Treatment at 50 degrees C produced a deposition approximately 8 and approximately 2.5 times higher than when treatments were carried out at 20 degrees C in IMZ and TBZ, respectively. No significant differences in terms of IMZ deposition were detected after treatments carried out alone or in combination. Uptake of the two fungicides was associated with their physicochemical characteristics as well as different formulation types. IMZ residues showed a great persistence during storage when applied separately, and >83% of active ingredient was present after 9 weeks of storage. IMZ residues increased with dip length, doubling when dip time increased from 0.5 to 3 min. In contrast, TBZ residues did not change with the different dip times. Following postharvest dip treatments of citrus fruit at 20 or 50 degrees C, home washing removed approximately 50% of the IMZ and approximately 90% of the TBZ.     

Occurrence of Azoxystrobin, Propiconazole, and Selected Other Fungicides in US Streams, 2005�C2006

Fungicides are used to prevent foliar diseases on a wide range of vegetable, field, fruit, and ornamental crops. They are generally more effective as protective rather than curative treatments, and hence tend to be applied before infections take place. Less than 1% of US soybeans were treated with a fungicide in 2002 but by 2006, 4% were treated. Like other pesticides, fungicides can move-off of fields after application and subsequently contaminate surface water, groundwater, and associated sediments. Due to the constant pressure from fungal diseases such as the recent Asian soybean rust outbreak, and the always-present desire to increase crop yields, there is the potential for a significant increase in the amount of fungicides used on US farms. Increased fungicide use could lead to increased environmental concentrations of these compounds. This study documents the occurrence of fungicides in select US streams soon after the first documentation of soybean rust in the US and prior to the corresponding increase in fungicide use to treat this problem. Water samples were collected from 29 streams in 13 states in 2005 and/or 2006, and analyzed for 12 target fungicides. Nine of the 12 fungicides were detected in at least one stream sample and at least one fungicide was detected in 20 of 29 streams. At least one fungicide was detected in 56% of the 103 samples, as many as five fungicides were detected in an individual sample, and mixtures of fungicides were common. Azoxystrobin was detected most frequently (45% of 103 samples) followed by metalaxyl (27%), propiconazole (17%), myclobutanil (9%), and tebuconazole (6%). Fungicide detections ranged from 0.002 to 1.15 ??g/L. There was indication of a seasonal pattern to fungicide occurrence, with detections more common and concentrations higher in late summer and early fall than in spring. At a few sites, fungicides were detected in all samples collected suggesting the potential for season-long occurrence in some streams. Fungicide occurrence appears to be related to fungicide use in the associated drainage basins; however, current use information is generally lacking and more detailed occurrence data are needed to accurately quantify such a relation. Maximum concentrations of fungicides were typically one or more orders of magnitude less than current toxicity estimates for freshwater aquatic organisms or humans; however, gaps in current toxicological understandings of the effects of fungicides in the environment limit these interpretations.     

Residue levels of ethoxyquin, imazalil, and iprodione in pears under cold-storage conditions.

A gas chromatographic method is presented for the simultaneous determination of the antiscald ethoxyquin and the fungicides imazalil and iprodione in peel and pulp of Blanquilla pears. Fruits were cold-stored in commercial chambers in normal atmosphere and in controlled atmosphere with low oxygen content (oxygen and carbon dioxide were held at 2.5% and 1.5%, respectively). The method uses gas-liquid chromatography (GLC) with an alkaline flame ionization detector (detector of N-P, NPD) and allows the detection of the mentioned compounds to minimum levels of 0.08-0.12 mg/kg in fresh fruit. With this system the evolution of residues in fruit was monitored throughout the period of cold storage. In the surveys carried out the residue levels of these compounds were found to be below the limits allowed by the legislation of European Union. For the three studied products residues in pulp are lower and disappear more quickly than in peel.     

哈密瓜常温保鲜贮运技术研究

该研究旨在解决新疆哈密瓜采后低温贮运费用高,常温贮运腐烂率高,损失严重的问题。针对哈密瓜采后腐烂的主要病原菌匍枝根霉(R.stolonifer)、链格孢霉(A.Alternata)、半裸镰刀菌(F.semitectum)、脂状青霉(P.digitatum),筛选出对症的抑菌剂,并研究其最适使用浓度;以常温贮藏哈密的瓜失重率、呼吸强度、过氧化氢酶(CAT)、纤维素酶、果胶甲酯酶(PE)、多聚半乳糖醛酸酶(PG)活性等为指标,研究云南虫胶作为被膜对哈密瓜采后生理的影响及其最适使用浓度。含有0.60g/L扑海因、0.8g/L氯硝胺、0.6g/L特克多、0.8g/L施宝功和1∶24(V.V-1)虫胶的混合溶液浸泡哈密瓜10s,于28℃,85%RH,贮藏30d,腐烂率1.87%,同期对照腐烂率49.54%。处理后的哈密瓜在室外温度28～34℃、棚车内温度30～38℃的条件下,经过12～15d由新疆哈密运抵苏州、上海、广州的腐烂率分别为3.37%、4.12%和4.86%。哈密瓜表皮以内0.8cm几乎无抑菌剂残留。用上述配方的虫胶与抑菌剂的混合溶液浸泡哈密瓜,可有效降低哈密瓜采后常温贮运中的腐烂率,保持良好风味和商品质量,大量节省运输费用     

Release of deuterated (E)-2-nonenal during beer aging from labeled precursors synthesized before boiling

Although lipid autoxidation in the boiling kettle is a key determinant of the cardboard flavor of aged beers, recent results show that mashing is another significant source of wort nonenal potential, the well-known indicator of how a beer will release (E)-2-nonenal during storage. Although unstable, deuterated (E)-2-nonenal nitrogen adducts created during mashing can in some cases partially persist in the pitching wort, to release deuterated (E)-2-nonenal during beer aging. In the experiment described here, the relative contributions of mashing and boiling were estimated at 30 and 70%, respectively. The presence of oxygen during mashing and, to a lesser extent, high lipoxygenase activity can intensify the stale cardboard flavor.     

Development of monoclonal immunoassays for the determination of triazole fungicides in fruit juices

Enzyme-linked immunosorbent assays (ELISAs) based on monoclonal antibodies for the detection of triazole fungicides have been developed. With this aim, hapten-protein conjugates, containing the common triazole and chlorinated aromatic moieties, were prepared. From mice immunized with these conjugates, several monoclonal antibodies (MAbs) with the ability to sensitively bind several triazoles with different specificity were obtained. Both analyte- and class-specific ELISAs were developed. The hexaconazole-specific immunoassay can determine this fungicide with a limit of detection of 0.3 mug/L in standard buffer. The so-called triazole-specific immunoassay allowed for the detection of tetraconazole, penconazole, cyproconazole, and myclobutanil, with limits of detection in the 0.1-0.7 mug/L range. These immunoassays were applied to the determination of triazoles in spiked fruit juices. Samples were adequately diluted to minimize the matrix effects. Coefficients of variation were below 30%, and recoveries ranged from 62 to 135%. Therefore, the developed immunoassays can determine triazole fungicides in fruit juices down to the maximum residue limits currently legislated, without any sample treatment other than dilution.     

Control of postharvest diseases of fruit by heat and fungicides: efficacy, residue levels, and residue persistence. A review

Extensive research has been done in recent years to reduce the heavy dependence on chemical fungicides to control postharvest diseases and disorders of horticultural crops. Alternative strategies were based on improved cultural practices, biological control, plant-defense promoters, and physical treatments such as UV illumination, radiofrequency treatment, heat therapy, and storage technologies. Among these, postharvest heat treatments such as hot water dips, short hot water rinsing and brushing, and hot air conditioning have reduced rot development and enhanced fruit resistance to chilling injury in sensitive cultivars while retaining fruit quality during cold storage and shelf life. Additive or synergistic increases in effectiveness were observed by integrating heat therapy with various chemical compounds, thus leading to significant reductions in the application of active ingredients to protect produce from decay. This paper highlights the knowledge on this topic with emphasis on heat therapy effects and factors affecting the uptake, persistence, and performance of fungicide residues when they are applied in combination with hot water.     

Fate of postharvest-applied dichlorvos in stored and processed dates

The fate of dichlorvos (DDVP) on dates after storage and processing of postharvest-treated fruits has been investigated. Residues were determined using GC-ECD after extraction of the fruits. A postharvest application was made to fruits at different stages of maturity: khalal fruits (mature full colored stage), rutab fruits (soft brown stage), and the mature tamer fruits (hard raisin-like stage). The fate of the residues was followed during the storage at refrigerated, room, and summer average temperatures (3, 22, and 43 degrees C). The amount of residues absorbed varied with the level of maturity. The rate of the loss of the residues was found to follow first-order kinetics. First-order rate constants were calculated for the different levels of maturity. The rate of the loss of DDVP increased as the temperature of storage increased. Also, it decreased with increasing maturation of the dates, which was characterized by a decrease in moisture content as well as water activity and an increase in sugar content. The period of storage study was limited by the time of maturation to the next stage. Most common home-cooking methods, including dehydration, jam-making, and syrup-making, resulted in significant decreases in residue levels. Only 0-13% of initial DDVP residues was detected in final products.     

Effect of pH on the solubilization of brewers' spent grain by microbial carbohydrases and proteases

The potential for enzymatic solubilization of brewers' spent grain by carbohydrases and proteases was examined over a broad pH range (pH 3.2-11.2). Enzymes from Trichoderma (Depol 686) were most efficient at a lower pH, while enzymes from the Humicola preparation (Depol 740) were the best performer over the whole range. Profiling of key glycoside hydrolase, esterase and protease activities across the pH range demonstrated that solubilization of spent grain by the Trichoderma enzymes corresponded to the range of maximum activities. This was not the case with the Humicola enzymes, where maximum solubilization of the substrate occurred at pH 9.1, at which pH the determined activities were low. Protease activity in Depol 740 was associated with a high solubilization, but inhibition of proteolytic activity resulted in only a 5% decrease in spent grain solubilization. These results suggest that while enzymes can be used to exploit agro-industrials byproduct, the use of high pH increases the extent of hydrolysis and an unidentified factor produced by Humicola improves the enzyme-catalyzed solubilization of lignocellulosic material.     

Dissipation of fungicides in a vineyard soil amended with different spent mushroom substrates

The degradation kinetics and formation of metabolites for fungicides of different chemical classes (iprovalicarb, metalaxyl, penconazole, and pyrimethanil) and determination of bound residues for metalaxyl and penconazole were studied in both an unamended vineyard soil and in the same soil amended with two spent mushroom substrates (composted (C-SMS1) and fresh (F-SMS2)). The degradation kinetics was fitted to single first-order or first-order multicompartment patterns. Degradation rates decreased in C-SMS1-amended soils for all fungicides as compared to unamended soil, but in F-SMS2-amended soils, they decreased only for iprovalicarb and penconazole. The DT(50) values were higher by up to 1.8 (metalaxyl), 3.8 (pyrimethanil), 4.1 (iprovalicarb), and >1000 (penconazole) times in the soil plus C-SMS1 compared to those for soil plus F-SMS2 or unamended soil. The dissipation mechanism recorded the highest mineralization in the unamended soil for (14)C-metalaxyl and (14)C-penconazole, with the highest formation of nonextractable residues in the F-SMS2-amended soil for (14)C-metalaxyl. The results are consistent with (1) the chemical characteristics of each SMS (total and soluble organic carbon) controlling sorption and the bioavailability of fungicides and (2) the microbial activity of SMS-amended soils, which affects fungicide biodegradation. The findings of this work highlight the potential of SMS amendments with different characteristics to decrease or increase the degradation rate of a fungicide in a vineyard soil.     

Storage Conditions Affecting Increase in Falling Number of Soft Red Winter Wheat Grain

Falling number (FN) of wheat grain, a measure of preharvest sprouting, tends to increase during storage; however, grain and storage conditions that impact FN changes are poorly understood. Wheat grain samples of varying FN from several cultivars were obtained by artificial sprouting, by incubating wheat stalks, or directly from the field and were used to determine the effects of cultivar, storage temperature, grain moisture content, and initial FN on changes in FN and α‐amylase activity. Increases in FN of artificially sprouted grain during storage were affected by temperature but not evidently by grain moisture in the range of 10.0–13.0%. The FN of artificially sprouted grain increased when stored at 5, 23, and 35°C for 20 weeks by averages of 9.4, 24.1, and 34.4, respectively. The influences of storage temperature and initial FN of grain obtained from incubated stalks were different between cultivars when stored for 8 weeks. Wheat grain obtained directly from the field also exhibited significant increases in FN during 8 weeks of storage at 5, 23, and 35°C with average increases of 10.0, 27.1, and 38.5, respectively. The impact of α‐amylase activity on the increase in FN during storage was evident only for field‐harvested grain of varying FN. α‐Amylase activity exhibited a negative logarithmic relationship (R² = 0.87) with FN in field‐harvested grain. The magnitude of the changes in α‐amylase activity varied by cultivar.     

Enzymatic and chemical treatment limits on the controlled solubilization of brewers' spent grain

The enzymatic hydrolysis of brewers' spent grain (BSG) has been investigated through treatment with commercial carbohydrases and proteases. Resultant residues were then chemically fractionated and delignified. Enzymatic treatments released 25-30% of the BSG mass and yielded precursors suitable for subsequent conversion to potentially value-added products. Controlled chemical fractionation selectively solubilized arabinoxylan but with no differences apparent due to prior enzyme treatment. The loss of non-polysaccharide components during alkali treatment suggests the presence of a high proportion of alkali-soluble lignin. Further delignification of the alkali-insoluble residues and further chemical fractionation released the remaining hemicellulose, to yield a residue which was >90% cellulose. Further knowledge of the properties and interaction between BSG polymers will facilitate an improved enzyme-assisted total deconstruction of BSG and hence the exploitation of its biomass.     

Residues of vinclozolin and procymidone after treatment of greenhouse grown lettuce,tomato and cucumber

Residues of the fungicides vinclozolin and procymidone, used to control the Botrytis cinerea disease, were studied in greenhouse grown lettuce, tomato and cucumber. Residue concentrations differed between the species of treated vegetables, and depended mostly on time of picking (harvest), size of fruits and mode of chemical application. The differences in concentration between one and two applications became smaller with time after application. The greatest differences occurred in lettuce. The residue contents in cucumber fruits assigned both for industrial and home processing (pickling) were higher than the legal limit (3 mg/kg). As a result of washing cucumbers, 22–24% of the residue was eliminated, and by peeling them 79–85% of the vinclozolin and procymidone residues were eliminated. Therefore it is reasonable that one or two applications of these fungicides should be used, with a time lapse of 14 days, but only with cucumbers intended for salad consumption. Vinclozolin residues, when used as vaporisation tablets, were in all cases 35–65% lower compared with the wettable powder (WP) formulation; however, the rate of of residue decrease was much slower. When applied in the form of vaporable tablets, the vinclozolin residue concentration was lower in all the sampled fruits in comparison with the WP formulation. The concentration of fungicide residues detected in winter tomato fruits was higher than that of the spring crop. Irrespective of vegetable the detected levels of fungicide residues were lower in ripe and bigger fruits than in green and smaller ones.     

Dissipation rates of cyprodinil and fludioxonil in lettuce and table grape in the field and under cold storage conditions

Two fungicides (cyprodinil and fludioxonil) have recently been used in southeast Spain to control disease in lettuce and grape. Gas chromatography with a nitrogen-phosphorus detector (GC-NPD) was used to study the disappearance of these compounds from crops under field conditions and during refrigeration. Residual values 21 days after application were below the maximum residue limit (MRL = 0.05 mg kg(-1)) established by Spanish law in the field experiment for both compounds. However, with the exception of fludioxonil in lettuce, residues were above the MRL in the refrigerated farm produce for both fungicides. The half-lives were 3-6 times greater under refrigeration.     

Effect of rice straw management on nitrogen balance and residual effect of urea-N in an annual lowland rice cropping sequence

Two field experiments were conducted in 1999 (wet season) and 2000 (dry season) on a Ustic Endoaquerts in central Thailand to examine the impact of rice straw management practices on rice yield, N uptake and fertilizer-N use efficiency. Treatments included a combination of urea broadcast at a rate of 70 kg N ha^у with either straw or compost which were incorporated at a rate of 5 Mg ha^у. At maturity of the wet season rice, ¹⁵N recovery by the grain was low (11-14%) as well as straw-N derived from labeled N (5-7%). After harvest, 25-29% of applied N still remained in the soil, mainly in the 0 to 5-cm layer. Large amounts of fertilizer-N (53-55%) were lost (unaccounted for) from the soil/plant system during the first crop. Residual fertilizer-N recovery in the second rice crop was less than 3% from the original application. During both fallow seasons NO₃^m-N remained the dominant form of mineral N (NO₃^m + NH₄⁺) in the soil but its concentration was low. In the wet season grain yield response to N application was significant (P =0.05). Organic material sources did not significantly change grain yield and N accumulation in rice. In terms of grain yield and N uptake at maturity, there was no significant residual effect of fertilizer-N on the subsequent rice crop. These results indicated that the combined use of organic residues with urea did not decrease total N losses or increase crop yield or uptake of N compared to urea alone.     

Flocculation and Haze Removal from Crude Beer Using In-House Produced Laccase from Trametes versicolor Cultured on Brewer's Spent Grain

The potential of brewer's spent grain (BSG), a common waste from the brewing industry, as a support-substrate for laccase production by the well-known laccase producer Trametes versicolor ATCC 20869 under solid-state fermentation conditions was assessed. An attempt was made to improve the laccase production by T. versicolor through supplementing the cultures with inducers, such as 2,2-azino bis(3-ethylbenzthiazoline-6-sulfonic acid), copper sulfate, ethanol, gallic acid, veratryl alcohol, and phenol. A higher laccase activity of 13506.2 ± 138.2 IU/gds (gram dry substrate) was obtained with a phenol concentration of 10 mg/kg substrate in a tray bioreactor after 12 days of incubation time. The flocculation properties of the laccase treated crude beer samples have been studied by using various parameters, such as viscosity, turbidity, ζ potential, total polyphenols, and total protein content. The present results indicated that laccase (25 IU/L) showed promising results as a good flocculating agent. The laccase treatment showed better flocculation capacity compared to the industrial flocculation process using stabifix as a flocculant. The laccase treatments (25 IU/L) at 4 ± 1 °C and room temperature have shown almost similar flocculation properties without much variability. The study demonstrated the potential of in-house produced laccase using brewer's spent grain for the clarification and flocculation of crude beer as a sustainable alternative to traditional flocculants, such as stabifix and bentonite.