Commensalism during submerged mixed culture of Geotrichum candidum and Penicillium camembertii on glutamate and lactate

Geotrichum candidum and Penicillium camembertii were cultivated in pure and mixed cultures on glutamate- and lactate-based medium. In pure culture, P. camembertii assimilated simultaneously glutamate, as a nitrogen and carbon source for biosynthesis, and lactate as an energy source. On the contrary, G. candidum grew on glutamate alone. The mixed culture led to higher growth rates and then higher rates of substrate consumption and metabolite production than each pure culture; however, the behaviour recorded was similar to that observed during G. candidum pure culture, in particular the absence of lactate assimilation during growth, illustrating a commensalism between both species. The presence of G. candidum induced a form of “competition” and then a better assimilation by P. camembertii of the sole nitrogen source, glutamate, which was therefore used as an energy source in addition to be a carbon (and nitrogen) source. Lactate was only used for energy supply during stationary state, as also recorded during G. candidum pure culture.     

Optimization of anaerobically digested distillery molasses spent wash decolorization using soil as inoculum in the absence of additional carbon and nitrogen source

The aim of this study was to achieve maximum decolorization of molasses spent wash (MSW) in absence of any additional carbon or nitrogen source using soil as inoculum. Soil samples were collected from the MSW disposal site. Colored soil samples exhibited higher pH, sugar and protein as compare to less colored samples. A decolorization of 69% was obtained using 10% (w/v) soil and 12.5% (v/v) MSW after 7 days incubation. Optimized parameters including days--6 days, pH--6, MSW--12.5% and soil concentration--40%, were obtained for maximum decolorization. A decolorization of 81% was achieved using 10% soil and 12.5% MSW after 18 days incubation in absence of any media supplement. Nearly 12% reduction in decolorization activity of the soil sample was observed over a period of 12 months when stored at 6 degrees C. It could be concluded that the decolorization of MSW might be achieved using soil as inoculum without addition of chemical amendments.     

亚热带红壤侵蚀区马尾松不同套种模式生态系统碳平衡

以福建省长汀县红壤侵蚀区马尾松低效林套种杨梅、无患子、油茶及黄栀子的改造模式林分为研究对象,对林分各组分生物量年净生长量、含碳率及土壤异养呼吸进行定位观测,分析套种模式对低效马尾松林分生态系统碳储量格局及碳平衡的影响。结果表明: 杨梅、无患子、油茶、黄栀子和马尾松不同器官含碳率的变化范围分别为41.1%～50.1%、42.2%～50.6%、45.1%～48.9%、44.7%～49.6%和46.1%～51.9%。不同树种同一器官之间的含碳率存在显著差异。马尾松套种杨梅及马尾松套种无患子模式植被层碳储量及年净增碳储量最高,分别为67.62～68.42 t·hm^-2和9.21～9.45 t·hm^-2·a^-1,马尾松套种油茶和马尾松套种黄栀子模式较小,分别为31.96～36.24 t·hm^-2和4.09～4.16 t·hm^-2·a^-1,马尾松纯林对照最小,分别为17.01 t·hm^-2和2.00 t·hm^-2·a^-1。土壤异养呼吸年通量从高到低依次为马尾松套种杨梅模式(7.41 t·hm^-2·a^-1)>马尾松套种油茶模式(5.89 t·hm^-2·a^-1)>马尾松套种无患子模式(5.86 t·hm^-2·a^-1)>马尾松套种黄栀子模式(4.95 t·hm^-2·a^-1)>马尾松纯林对照(2.45 t·hm^-2·a^-1)。马尾松套种杨梅和马尾松套种无患子模式的年净生态系统碳平衡分别为2.04和3.27 t C·hm^-2·a^-1,表现为“碳汇”,马尾松套种油茶和马尾松套种黄栀子模式及马尾松纯林对照的年净生态系统碳平衡分别为-1.80、-0.80和-0.45 t C·hm^-2·a^-1,表现为“碳源”。总体上,短期内马尾松低效林套种杨梅或无患子能够提升林分的固碳增汇效益。     

AM真菌控制蚕豆枯萎病发生的根际微生物效应

在温室盆栽条件下,研究了接种尖孢镰刀菌蚕豆专化型和丛枝菌根(AM)真菌(摩西球囊霉Gm、扭形球囊霉Gt、根内球囊霉Gi及幼套球囊霉Ge)对灭菌连作土壤中蚕豆幼苗生长、枯萎病发生、根际镰刀菌数量和微生物代谢功能多样性的影响.结果表明: 接种AM真菌能显著增加蚕豆幼苗地上部和地下部鲜质量;接种Gm、Gt、Gi和Ge真菌使蚕豆枯萎病病情指数分别显著降低94.0%、60.0%、64.0%和94.0%,使蚕豆根际镰刀菌数量分别显著降低98.6%、74.3%、77.8%和90.4%,以Gm和Ge真菌对蚕豆枯萎病的抑制效应最好.接种Gm、Gt和Ge显著提高了根际微生物对糖类(CH)、氨基酸类(AA)、羧酸类(CA)和酚酸类(PA)碳源的利用,使蚕豆根际微生物的AWCD值分别显著提高34.4%、31.5%和50.8%;而接种Gi对AA、CA、PA类碳源利用和AWCD值均无显著影响,使微生物对CH类碳源的利用显著降低59.3%.主成分和相关分析表明, 接种AM真菌明显改变了蚕豆根际微生物的群落结构.接种Gt真菌提高了蚕豆根际微生物对AA类碳源(β-甲基-D-葡萄糖苷、D-半乳糖酸-γ-内酯、D-甘露醇、N-乙酰基-D-葡萄糖胺、D-纤维二糖)和CA类碳源(D-半乳糖醛酸)的利用,而接种Gi真菌降低了微生物对以上碳源的利用;接种Gm和Ge真菌提高了根际微生物对L-精氨酸和4-羟基苯甲酸的利用.表明糖类和羧酸类是接种Gt真菌后蚕豆根际土壤微生物利用的主要碳源, 氨基酸和酚酸类是接种Gm和Ge后蚕豆根际土壤微生物利用的主要碳源.接种AM真菌显著提高了蚕豆根际微生物的活性,改变了微生物群落结构并抑制病原菌的增殖,是AM真菌减轻了蚕豆枯萎病的发生的重要原因.不同AM真菌减轻了蚕豆枯萎病的发生与其改变微生物的碳源利用密切相关.     

Prey-Dependent Life Attributes of an Aphidophagous Ladybird Beetle, Propylea dissecta (Coleoptera: Coccinellidae)

Predation potential, development, immature survival and reproduction of an aphidophagous ladybeetle, Propylea dissecta (Mulsant) was studied when fed on seven aphid prey, viz. Aphis gossypii, Aphis craccivora, Lipaphis erysimi, Uroleucon compositae, Brevicoryne brassicae, Rhopalosiphum maidis and Myzus persicae. A. gossypii was most suitable and consumed by the larvae and adults of P. dissecta, while M. persicae, the least. Pre-imaginal development of P. dissecta was fastest (0.080 day^-1) when A. gossypii was used as prey, whilst slowest (0.061 day^-1) on M. persicae. The immature survival, adult emergence, adult male and female longevity of P. dissecta was maximal (i.e., 77.10±0.04 and 93.21±0.79%, 57.10±1.62 and 62.40±1.93 days, respectively) on A. gossypii and minimal (i.e., 63.01±1.87 and 81.73±1.79%, 42.50±1.21 and 49.40±2.32 days, respectively) when M. persicae was provided as prey. Oviposition period, fecundity, percent egg viability and mean reproductive rate was maximum (i.e., 50.30±2.03 days, 856.00±30.00 eggs, 96.40±0.31% and 17.02 eggs per day) on A. gossypii, and minimum (i.e., 18.00±1.40 days, 212.00±18.21 eggs, 72.46±2.81% and 11.78 eggs per day) on M. persicae. Adult weight and developmental rate of P. dissecta have a positive correlation, which suggests that if immature stages of ladybeetle developed faster, they should grow into heavier adults. Female longevity and fecundity also have a positive correlation. The findings also reveal that all seven aphid species tested are essential food. Rank order of prey species was consistent in all experimental parameters.     

Critical thermal maximum and body water loss in first instar larvae of three Cetoniidae species (Coleoptera)

Critical thermal maximum (CT_max) and body water losses were measured in first instar larvae of Gnorimus nobilis, Osmoderma eremita (Trichiinae) and Cetonischema aeruginosa (Cetoniinae) when air temperature was increased gradually (0.5 °C/min) from 20 °C to the critical thermal maximum (CT_max), in dry air (near 0% R.H.).

The CT_max was significantly lower in O. eremita (45.6±0.7 °C) than in G. nobilis (48.5±0.6) and C. aeruginosa (51.4±0.9 °C).

An increase of 10 °C (30–40 °C) induced a 2-fold increase of the water loss in C. aeruginosa and O. eremita (Q₁₀=2.10±0.12 and 2.13±0.20, respectively). In the range from 40 to 45 °C to CT_max a strong increase of the water loss was observed in O. eremita and C. aeruginosa, respectively. Body water losses were significantly lower in C. aeruginosa than in O. eremita and G. nobilis over the range 20 °C—CT_max; no significant difference occurred between G. nobilis and O. eremita.     

Bioremediation and decolorization of anaerobically digested distillery spent wash

A facultative anaerobic pure bacterial culture L-2 capable of growth on 12.5% (v/v) diluted digested spent wash supplemented with glucose (10g/l) was isolated from an Indian distillery. It achieved 31% decolorization and 57% COD reduction after 7 days' incubation. The advantages of using such a culture for digested spent wash bioremediation are apparent in providing a realistic approach for decreasing its pollution potential prior to disposal.     

Chemical structure and antiviral activity of carrageenans from Meristiella gelidium against herpes simplex and dengue virus

Chemical and spectroscopic methods showed that the major KCl-precipitated galactans from Meristiella gelidium (Solieriaceae) are iota/kappa/nu-hybrid carrageenans with the former one in higher proportion. These carrageenans showed, by HPSEC-MALLS analysis, unimodal symmetrical peaks with MW of 425.6–956.7 kDa. The effectiveness of the crude extracts from M. gelidium against HSV-2 was higher than the corresponding extract from G. griffithsiae, previously determined. However, when considering the homogeneous carrageenans, the fractions obtained from both seaweeds showed the same level of activity. The extracts and carrageenan derived from M. gelidium were more effective inhibitors of DENV-2 if compared with G. griffithsiae samples and reference polysaccharides. The most active fraction obtained from M. gelidium showed a selectivity index against HSV-2 of 25,000, a value high enough to consider this carrageenan as a promising agent to be evaluated for the treatment of genital HSV-2 infections.     

Non-Ligninolytic TNT Mineralization in Contaminated Soil by Phanerochaete chrysosporium

The explosive 2,4,6-trinitrotoluene (TNT) is widely used and results in widespread soil contamination. The white-rot fungus Phanerochaete chrysosporium has been shown to degrade TNT, using the peroxidase enzyme. In this study, we report peroxidase-independent degradation of TNT by non-ligninolytic P. chrysosporium. Significant disappearance of TNT from highly contaminated soil using P. chrysosporium has been observed. Soil highly contaminated with TNT (2270 ppm [10 mM]) was diluted to 100 ppm (0.44 mM) with malt extract medium. Pregrown (48 hours) mycelial pellets of P. chrysosporium were added in 100 mL malt extract medium and incubated in Gledhill flasks. Analysis by high-performance liquid chromatography (HPLC) was conducted on soil extracts at specific time points to estimate the disappearance of TNT from contaminated soil incubated with P. chrysosporium. When the pregrown mycelial pellets were added, TNT disappeared within 48 hours. The dissolved concentration of 2-amino-4,6-dinitrotoluene (2Am-DNT) increased up to the third day, then declined before its final disappearance by day 10. Results show that the pregrown mycelial pellets of P. chrysosporium mineralized up to 17.3±6.3% [¹⁴C]-TNT within 30 days.     

A Study of the Stereocontrolled Reduction of Aliphatic β-Ketoesters by Geotrichum candidum

The conditions necessary to reduce stereoselectively aliphatic beta-ketoesters by G. candidum have been investigated. Ageing of the mycelium allowed a practical preparation of optically pure (R)-ethyl 3-hydroxybutanoate, as a result of stereoselective reduction, enantioselective metabolism and stereoisomer interconversion. The stereoconversion of 3S-hydroxybutanoate into the 3R-enantiomer via 3-oxoester formation has been demonstrated to be a determining factor in the building of the optical purity of the resulting product. Enantioselective preparations of higher homologous 3R-hydroxyesters are described.     

Enhanced activity and stability of immobilized lipases by treatment with polar solvents prior to lyophilization

Lipases from Candida rugosa, Mucor javanicus and Rhizopus oryzae were respectively adsorbed on Amberlite XAD-7 followed by incubation in 2-propanol and then lyophilization. The activities of the immobilized enzymes were 1.6–3.4 times higher than those of the immobilized enzymes without incubation in the organic solvent before lyophilization for esterification of lauric acid (0.1 M) and 1-propanol (0.1 M) in isooctane at 37 °C. The immobilized C. rugosa lipase (Sigma) without the incubation did not show any activity but displayed considerable activity (19.8 μmol h⁻¹ mg⁻¹) after the incubation before lyophilization. Besides 2-propanol, acetone, 1-propanol and ethyl acetate were also found to be good solvents for treating M. javanicus lipase immobilized on Amberlite XAD-7 and acetone was the best among them. When incubated in isooctane at 25 °C for 120 h, the immobilized M. javanicus lipase prepared by incubation in acetone for 1 h before lyophilization retained 70% of its initial activity while the immobilized enzyme without the solvent treatment kept only 50% of its initial activity.     

Preferred temperatures of desert honey ants (hymenoptera: formicidae)

1. 1.Preferred temperature ranges were determined for workers of 5 species of Myrmecocystus in a temperature gradient on wet sand.

2. 2.Workers of the diurnally foraging species, M. mendax Wheeler, M romainei Cole, M. depilis Forel, and M. mimicus Wheeler, were distributed between 7 and 45°C, and workers of one nocturnal species, M. mexicanus Wesmael, were distributed between 5 and 35°C.

3. 3.Mean preferred temperatures of the 5 species increased as follows: mexicanus < mendax < romainei < depilis < mimicus.

4. 4.Preferred temperatures of M. romainei and M. mexicanus were higher in the summer than in the fall.

5. 5.After 2 h in the gradient on dry sand, workers of M. romainei were distributed similarly to those on wet sand, but after 8 h on dry sand, most workers were below 12°C where the air was saturated.

Removal of chlorophenols from synthetic solutions using Phanerochaete chrysosporium

The potential use of the fungus Phanerochaete chrysosporium to remove chlorophenols (phenol, o-chlorophenol, p-chlorophenol and 2,4,6-trichlorophenol) from aqueous solutions was evaluated. The kinetics of both adsorption and desorption of phenolic compounds was rapid for all adsorbates. The maximum adsorptions of phenol and chlorophenols onto the Phanerochaete chrysosporium were 1.23 mmol/g for phenol, 1.49 mmol/g for o-chlorophenol, 1.78 mmol/g for p-chlorophenol and 2.14 mmol/g for 2,4,6-trichlorophenol. The affinity order was as follows: 2,4,6-trichlorophenol > p-chlorophenol > o-chlorophenol > phenol. Phenol and chlorophenols binding with Phanerochaete chrysosporium were clearly pH dependent. The adsorption of phenol and chlorophenols increased as pH increased. Desorption of phenol or chlorophenols was achieved using methanol solution (30% (v/v)). Phanerochaete chrysosporium is suitable for reuse for more than ten cycles without noticeable loss of adsorption capacity.     

Synergistic effects of antagonistic fungi and a plant growth promoting rhizobacterium, an arbuscular mycorrhizal fungus, or composted cow manure on populations of Meloidogyne incognita and growth of tomato

Effects of four antagonistic fungi (Paecilomyces lilacinus, Pochonia chlamydosporia, Trichoderma harzianum and Gliocladium virens) alone and together with a plant growth promoting rhizobacterium Pseudomonas putida, an arbuscular mycorrhizal fungus Glomus intraradices or with composted cow manure (CCM) were assessed on the growth of tomato and on the reproduction of Meloidogyne incognita in glasshouse experiments. Application of all antagonistic fungi (except G. virens), P. putida, G. intraradices or CCM caused a significant increase in the growth of plants without nematodes. However, use of either of these fungi, P. putida, G. intraradices and CCM against plants with nematodes caused a significant increase in tomato growth. Paecilomyces lilacinus caused a 42% increase in the growth of nematode-inoculated plants followed by P. chlamydosporia (36%), T. harzianum (18%) and G. virens (15%). CCM caused about 57% increase in the growth of nematode-inoculated plants followed by P. putida (37%) and G. intraradices (31%). Maximum increase (71%) in the growth of nematode-inoculated plants was observed when CCM was used with P. lilacinus. Moreover, P. lilacinus caused a high reduction (55%) in galling and nematode multiplication, while G. virens the least (25%). Use of P. putida also caused a 39% reduction in galling and nematode multiplication followed by CCM (34%) and G. intraradices (32%). Combined use of CCM with P. lilacinus caused maximum reduction (79%) in galling and nematode multiplication. Re-isolation of antagonistic fungi from nematodes revealed that P. lilacinus parasitised more females and eggs than other antagonistic fungi. Root colonisation by P. putida was increased with P. lilacinus, while colonisation by G. intraradices was reduced in the presence of antagonistic fungi.     

Effect of different operational conditions on the decolorization of molasses spent wash using once developed soil inoculum

A stirred vessel coupled with membrane unit containing cellulose acetate (0.45 μm) membrane was used to study the decolorization of anaerobically digested molasses spent wash (MSW). The soil collected from the MSW disposal site was used as inoculum to study the decolorization without addition of any additives. The same inoculum was used over a period of 163 days at room temperature to study the decolorization of 12.5–50% (v/v) MSW using different operational conditions. The reactor was entered in to the inhibition mode after the feeding of 50% MSW, which was restored 100% without changing any operational condition. The maximum decolorization obtained for 12.5% (v/v) MSW was 77.22 ± 0.13%. The decolorization achieved for 25, 37.5, and 50% (v/v) MSW was 70.41 ± 0.12, 56.47 ± 0.17, and 48.78 ± 0.09%, respectively. Increase in the utilization of protein and reducing sugar was observed up to 25% MSW whereas, higher concentration showed decrease in the utilization. Results indicate 63% removal of chemical oxygen demand for 12.5% (v/v) MSW. Membrane flux which was significantly reduced after the feeding of 50% MSW was regenerated without changing the washing procedure, however, 35% decrease in sample flux was observed over the continuous use of membrane for the period of 198 days.     

Flavonoid levels in roots ofMedicago sativa are modulated by the developmental stage of the symbiosis and the root colonizing arbuscular mycorrhizal fungus

Abundant data on the effect of flavonoids on spore germination, hyphal growth and root colonization by AMF are available. Moreover, the flavonoid pattern in mycorrhizal roots changes, thus flavonoids have been suggested as arbuscular mycorrhizal signalling compounds. In our work we studied the accumulation of flavonoids in roots of Medicago sativa i) after the exposure of uncolonized roots to sterile solutions containing Glomus intraradices tissue, ii) at three different stages of colonization by G. mosseae, iii) colonized by G. mosseae, G. intraradices or Gigaspora rosea.

We could show that flavonoid accumulation in M. sativa roots i) is induced before root colonization, pointing towards the presence of a fungal-derived signal, ii) depends on the developmental stage of the symbiosis and iii) depends on the root-colonizing arbuscular mycorrhizal fungus. The data presented indicate not only a time-specificity of the flavonoid accumulation during the mycorrhizal association, but also an arbuscular mycorrhizal fungal-specificity. The possible functions of the flavonoid pattern changes are discussed.     

Gelling properties of water-soluble polysaccharides from proliferating marine green seaweeds (Ulva spp.)

Water-soluble polysaccharides (12·2% of the algal dry weight) were extracted from marine green seaweed (Ulva spp.) which proliferate along the Brittany shores of France. They were composed of 18·4% rhamnose, 4·4% glucose, 1·9% xylose, 0·9% mannose, 0·9% galactose, 15·2% uronic acid, 15·8% sulphate and 23·7% ash based on the extract dry weight. These polysaccharides, formed a weak gel (about 3 Pa) at a concentration of 1·6% (w/v) in deionized water. The elastic modulus increased to about 160 Pa when boric acid (15–33 m ) was added and reached 250 Pa when both boric acid (7 m ) and calcium chloride (7 m ) were present. Adjusting the pH to 7·5 or higher by sodium tetraborate, phosphate or Tris-HCl buffers was detrimental to the gel. These results demonstrate that the poorly exploited biomass of Ulva spp. is a source of gelling polysaccharides of potential economical value. Mechanisms for gel formation which unusually involve both boron and calcium ions are proposed and will be studied further.     

Purification and partial characterization of manganese peroxidase from immobilized Phanerochaete chrysosporium

Solid-state culture of the white-rot fungus Phanerochaete chrysosporium BKMF-1767 (ATCC 24725) has been carried out, using an inert support, polystyrene foam. Suitable medium and culture conditions have been chosen to favor the secretion of manganese peroxidase (MnP). The enzyme was isolated and purified from immobilized P. chrysosporium and partially characterized. Partial protein precipitation in crude enzyme was affected using ammonium sulphate, polyethylene glycol, methanol, and ethanol methods. Fractionation of MnP was performed by DEAE-Sepharose ion exchange chromatography followed by Ultragel AcA 54 gel filtration chromatography. This purification attained 23.08% activity yield with a purification factor of 5.8. According to data on gel filtration chromatography and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), the molecular weight of the enzyme was 45 000±1000 Da. The optimum pH and temperature of purified MnP were 4.5 and 30 °C, respectively. This enzyme was stable in the pH range 4.5–6.0, at 25 °C and also up to 35 °C at pH 4.5 for 1 h incubation period. MnP activity was inhibited by 2 mM NaN₃, ascorbic acid, β-mercaptoethanol and dithreitol. The K_m values of MnP for hydrogen peroxide and 2.6-dimetoxyphenol were 71.4 and 28.57 μM at pH 4.5, respectively. The effects of possible inhibitors and activators of enzyme activity were investigated.     

外部干扰对根茎型克隆植物甘草自然种群植株生长及主要药用成分含量的影响

克隆植物, 特别是游击型克隆植物, 其分株在一定时间段往往通过连接结构(即间隔子)连接在一起形成克隆网络, 基于克隆网络的克隆植物特性显著提高了克隆植物应对外部干扰的能力。长期封育禁牧使得甘草(Glycyrrhiza uralensis)自然种群免受外部干扰, 却并没有促使受破坏的甘草自然种群快速恢复。因为克隆整合、克隆储存及克隆分株的选择性放置等克隆植物特性的存在, 适度的外部干扰有利于根茎型克隆植物甘草的生长与繁衍。但外部干扰与克隆植物特性在甘草种群恢复中的作用及其机理目前并不清楚。该研究以甘草自然种群为研究对象, 通过野外控制实验进行模拟动物采食和人工采挖干扰, 探讨不同程度干扰对甘草自然种群生长和主要药用成分含量的影响。结果表明在中度放牧采食干扰或者人工采挖干扰下, 甘草自然种群的分株密度、高度和地上生物量与对照相比均没有显著差异, 甘草表现出等补偿生长; 而在不同类型的重度干扰下, 甘草产生的响应不一致。重度放牧采食干扰下甘草表现出欠补偿生长, 而重度人工采挖干扰下甘草表现出超补偿生长。克隆植物特性在甘草应对外部干扰过程中发挥着重要作用。外部干扰一定程度上提高了甘草中甘草苷和甘草酸的含量, 从而提升了药材的品质。进一步的研究应该关注于甘草自然种群与环境因子之间的关系, 解析甘草自然种群退化的原因, 提出相应的复壮技术, 探讨可持续利用的管理模式。     

Phenols removal in musts: Strategy for wine stabilization by laccase

The potential of laccase from Trametes versicolor for phenolic removal in must for wine stabilization was evaluated through a combination of an analytical methodology (capillary zone electrophoresis) and kinetics of phenols removal as the total antioxidant potential variation. Total phenolic content, total antioxidant potential and polyphenols were monitored from 0 to 3 h of must treatment. The results indicated that the treatment of a red must with laccase affect mainly the phenolic compounds responsible for the must antioxidant properties. The treatment of white musts with laccase showed higher reduction in total phenol than in the total antioxidant potential. Phenol degradation by laccase was very fast for catechins, and slowly for stilbenes (cis- and trans-resveratrol) and derivatives of cinnamic (ferulic and caffeic) and benzoic (syringic, vanillic, and gallic) acids. It is possible to conclude in this case that the use of laccase in white wines is perfectly feasible. This would allow softer and ecologically correct treatments, which would diminish the cost of processing and avoid deterioration of wines for long storage times.