FI57875B - ANTIFUNGALT PREPARAT - Google Patents

Description

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lU.02.75 Great Britain-Storbritannien (GB) 29503/7 ^, 061 + 50/75 (71) Gist-Brocades nv, Wateringseveg 1, Delft, Netherlands-Nederländerna (NL) (72) Dirk Aart Smink, Leiderdorp, The Netherlands This invention relates to an antifungal preparation for use in agricultural products such as fruit, vegetables, root vegetables, beets, flower bulbs and rhizomes.

Citrus fruits, such as oranges in particular, but also lemons and grapefruits, are exported in large quantities from the countries where they are grown to many other countries. As a result, they have to be transported for long periods of time in all kinds of conditions, many of which cause fruit spoilage due to mold growth. In addition, fruit is often stored for long periods of time before it reaches the consumer, which also contributes to the conditions under which mold growth occurs.

To avoid spoilage, citrus fruits are therefore usually treated with an antifungal agent before transport. However, despite treatment of citrus fruits with an antifungal agent (or fungicide), mold growth often occurs to the extent that 57875 a significant proportion of citrus fruits cannot be sold upon arrival in the recipient country, and for example one moldy orange can have a devastating effect on all surrounding fruits.

It has already been discovered that natamycin (pimaricin, see Merck Index, 8th Edition (1968), page 834) is a highly active antifungal agent. However, for citrus processing, it is necessary to apply sufficient amounts of natamycin to the citrus peel, especially if the infection occurred several days before processing, as is the case in most cases. The use of a natamycin suspension provides insufficient penetration into the peel of citrus fruits and thus insufficient protection against mold growth. We have found that by extending the immersion times of the fruit to at least one hour, the anti-mold efficacy of natamycin could be increased, but this application is still quite impractical and uneconomical. On the other hand, we have found that a solution of natamycin in dimethyl sulfoxide (DMSO) gives the desired effect against mold growth, but the odor of DMSO is therefore permeable that this solution is unsuitable for practice. We have also found that the methyl ester of natamycin as an adduct with undecylenic acid has some improved efficacy, but penetration into the citrus peel is still insufficient for applications due to an apparently unfavorable hydrophilic-lipophilic balance.

We have now found that a very useful and effective preparation can be obtained by combining natamycin or another polyene antibiotic with an alcohol and an organic acid. In such a mixture, the solubility of natamycin or another polyene antibiotic is sufficient to penetrate the peel of the citrus fruit to such an extent that the growth of mold is virtually completely inhibited even during prolonged transport and storage, which are not always unavoidable. Also, if there is mold growth in the citrus fruit, then spore formation is strongly inhibited, so mold growth occurs only locally without the risk of whole boxes of citrus fruits starting to rot.

It has further been found that the above preparation can be used with equal advantage for other agricultural products, such as other fruits, vegetables, root crops, beets, flower bulbs and rhizomes. As with citrus fruits, penetration into the skin or wall of other agricultural products is clearly better than with the compositions presented so far.

According to the present invention, there is thus provided a preparation for treating agricultural products such as fruit, vegetables, roots, beets, flower bulbs and rhizomes, and the preparation of the invention is characterized in that it contains about 0.04-0.2% of a polyene antibiotic containing natamycin. , lucensomycin or aureofungin, dissolved in a mixture containing about 0.1-1% alkanoic acid having 1-8 carbon atoms, 1-3 carboxyl groups and 0-2 hydroxy groups, and 10% to the remainder of the mixture, preferably 15-50% alkanol with 1-3 carbon atoms.

The preferred polyene antibiotic is natamycin or a salt or ester thereof, for example a sodium salt or a methyl ester. The solubility of natamycin in the mixture of lower alkanol and lower alkanoic acid is 500-1000 ppm, and the mixture shown is sufficient to treat said agricultural products to prevent mold effectively and for a long time.

The term "lower" as used herein, in connection with alkanols and alkanoic acids, means that the alkanol or alkanoic acid has at most 8 carbon atoms, preferably 2 or 3 carbon atoms, the carbon atoms may be straight or branched chain, the alkanol may contain at most one hydroxy group per carbon atom, and preferably has one hydroxy group in the molecule Preferred alkanols are methyl, ethyl, and propyl alcohols Alkanoic acids may contain 1-2 carboxy groups Preferred alkanoic acids include acetic acid and propionic acid The alkanoic acid may contain substituents such as 1-2 hydroxy groups, e.g., lactic acid, and / or one or more halogen atoms, preferably chlorine atoms, e.g. chloroacetic acid and trichloroacetic acid.Other more complex acids may also be used, e.g. citric acid and ascorbic acid. no residues are left on agricultural products which are harmful and / or unpleasant when the products are used.

57875

The preparations according to the invention may also contain water and additives, such as coating agents, e.g. plastic emulsions, such as polyvinyl acetate emulsions, antioxidants, e.g. ascorbic acid, wetting agents and thickeners, such as gelatin.

Examples of agricultural products that can be treated with the preparations of this invention include fruits such as citrus fruits, e.g., oranges, lemons and grapefruits, other fruits such as bananas, avocados, pajamas, mangoes, lychees, melons, apples, pears, plums, cherries, peaches. , grapes, tomatoes and cucumbers, vegetables such as cabbage and celery, root crops such as potatoes, sweet potatoes, turnips, celery, radishes, onions and dahlias, beets such as carrots and sugar and white beets, onions, onions, tulips , gladiolus bulbs, hyacinth bulbs and iris bulbs, and rhizomes.

The preparations according to the invention can be used as baths in which agricultural products are immersed. Short immersion times, preferably 1 to 10 minutes, more preferably 1.5 to 3 minutes, are perfectly suited to provide agricultural products with lasting protection against mold. Agricultural products can be immersed directly in the solution. According to another method, the preparations are applied to agricultural products by brushing with brushes; in this case, for example, rotating brushes can be mounted on a somewhat inclined surface, and agricultural products are passed over the brushes. The brushes are moistened, for example, by allowing their wheels to pass through the surface of the preparation. According to yet another method, the preparations can be sprayed with agricultural products.

The preparations according to the invention can be easily prepared by dissolving a polyene antibiotic, for example natamycin, in a lower alkanol and a lower alkanoic acid in the customary manner.

The invention is illustrated by the following examples.

5,57875

Example 1

The experiments were performed on oranges. The peel of the oranges was scratched and the oranges were immersed for 15 seconds in a suspension of about 150,000 spores / ml (a spore mixture of the molds of Penicillium digitatum and P. italicum commonly found in oranges), and the oranges were then allowed to dry overnight.

The oranges thus treated were dipped in the solutions shown below for 2 minutes, then stored for 7 days at 26 ° C and 95% relative humidity. 10 oranges were used for each experiment, and a further 10 oranges were used for the control. The results were as follows: Treatment Number of oranges with mold growth control 10 methyl alcohol + 0.1% lactic acid + 0.1% natamycin 1 water + 0.1 55 lactic acid + 0.1% natamycin 6 water + 0.1% Mertect 340 "(" Mertect "is a trademark of 2,4-thiazolylbenzimidazole, also known as thiobendazole or TBZ) 6

It is obvious that the best prevention is obtained with a solution of natamycin in alcohol and organic acid.

Example 2 In this example the so-called "Jaffa" oranges, and were infected in the same manner as in Example 1, except using a suspension containing 120,000 spores / ml. The storage conditions were also the same as in Example 1. The following results were obtained: 6 57875 Treatment Number of oranges with mold growth control 10 methyl alcohol + 0.1 ί lactic acid + 0.1% natamycin 0 ethyl alcohol + 0.1% lactic acid + 0.1% natamycin 1 n-propyl alcohol + 0.1 / 5 lactic acid + 0.1% natamycin 1 methyl alcohol + 0.1% acetic acid + 0.1% natamycin 0 methyl alcohol + 0.1% propionic acid + 0.1% natamycin 0 water + 0 , 1¾ "Mertect 3 ^ 0" 8 This example shows that good results are obtained using several different alcohols and several different acids.

Example 3 Jaffa oranges were used in this example and were infected as described in Example 1. The storage conditions were also otherwise the same as in Example 1, except that the storage time was 8 days. The following results were obtained: Treatment Number of oranges with mold growth control 8 ethyl alcohol-water (1: 1) + 0.1% acetic acid + 0.1 5 natamycin 6 ethyl alcohol-water (1: 1) + 0.5% acetic acid + 0.1 55 natamycin 3 ethyl alcohol-water (1: 1) + 1.0 55 acetic acid + 0.1 55 natamycin 2

The results show that increasing the acid concentration provides better prevention of mold growth.

Example U

In this example, the immersion treatment was compared to the brushing treatment. The oranges were infected as described in Example 1, except that a suspension containing 120,000 spores / ml was used. The storage conditions were the same as in Example 1, except that the storage time was 10 days. 20 oranges were used in each experiment.

Treatment Number of oranges with mold growth control 13 immersion for 2 minutes in: ethyl alcohol-water (1: 1) + 1% acetic acid + 0.1% natamycin 2 brushed with: ethyl alcohol-water (1: 1) + 1 K * acetic acid + 0.2 55 natamycin x '2 brushed with a mixture of: s water + 0.2 $ "Seas 340nX' 8 This example shows that the brushing treatment gives the same result as the immersion treatment when twice the amount of antibiotic is used for the brushing treatment. The preparations according to the invention also gave clearly better results than the known "Mertect" at the same concentration.

X) these solutions also contained 10% polyvinyl acetate solution. Example 5 Untreated lemons were scratched and then immersed for 15 seconds in a suspension of about 900,000 spores / ml. The lemons were allowed to dry overnight. The dried lemons were brushed with the preparations shown below. The storage conditions were otherwise the same as in Example 1, except that the storage time was 17 days instead of 7.

Treatment Number of lemons with mold growth 30% ethyl alcohol + 1% acetic acid + 10% polyvinyl acetate 7 same + 0.2% natamycin 2 same + 0.2% aureofungin 1 same + 0.2% lucensomycin 2 This table shows that the polyene antibiotics disclosed provide good protection against mold growth.

5 7 8 7 5 8

Example 6

Freshly harvested banana bunches were washed under running water and dried. The bananas were then treated with the preparations shown below by immersing them in them for about 15 seconds, then allowed to drain and packed after drying in cardboard boxes. The boxes were kept at 12.5 ° C for 14 days, and the bananas were then ripened for 7 days. The degree of stem rot in each bunch was estimated to be zero (no fungal growth) or 1-4, with numbers indicating an increasing degree of fungal growth. For this treatment, 6 boxes were used, each with 8 bunches.

Treatment Mushroom rot,% in the body of the bunch in the banana stem water 46 22 30% ethyl alcohol + 0.5% acetic acid + distilled water to 100 5S 44 23 30% ethyl alcohol + 0.5% acetic acid + 0.2 K natamycin + distilled water $ 100: ksi 8 7 water + 0.2% natamycin 34 18 This example clearly shows an improvement in mold prevention when alcohol and an organic acid are used in the preparation.

Example 7

The oranges were scratched and then immersed in a suspension of 500,000 spores / ml for 15 seconds. Infected oranges were allowed to dry overnight. The oranges were then brushed with the solutions shown below and stored for 6 weeks at 18 ° C and $ 95 relative humidity. Fourteen oranges were used for each experiment. The results are as follows: Treatment Number of spoiled fruits control 12 propanol-water (3: 7 v / v) + 1% gelatin + 1 i acetic acid 10 propanol-water (3: 7 v / v) + 1 Ϊ gelatin + 1% acetic acid + 0.2% natamycin 0 propanol-water (2: 8 v / v) + 1% gelatin + 1% acetic acid li 9 57875 propanol-water (2: 8 v / v) + 1% gelatin + 1% acetic acid + O ', 2% natamycin 2 $ 0.2 "Mertect 340" 12 0.2% natamycin in water 13 This example shows that good results are obtained even after longer storage times when the antifungal preparations used contain alcohol and an organic acid.

Example 8

Potato disinfection A. Rhizoctonia solani * 11a infected potatoes were left to germinate in the dark at room temperature. The sprouts were drilled from the potatoes with a cork drill and, after drying, the pieces were dipped in molten paraffin to protect the pieces from the outside. The 50 pieces were then treated for 30 minutes with the disinfectant solutions shown in the following table. After treatment, the pieces were dried and implanted in sterile soil. The seedlings were allowed to grow for 21 days at 20 ° C and 90-95% relative humidity. After this time, the sprouts were examined for early disease (sprouts completely rotted) and subsequent infection (mycelium and / or small brown lesions in the sprouts).

% suspended or dissolved in water:% of germs affected: natamycin ethyl alcohol acetic acid early late not affected 0.1 50 0.1 2 8 84 50 0.1 24 60 14 - 54 30 16 0.1 - - 18 42 40

The table shows that the disinfection of potatoes with natamycin in the presence of ethyl alcohol and acetic acid is clearly better than with natamycin alone.

B. Mycelium cut from potatoes infected with Rhizoctonia solani * 11a was used in this experiment. 100 mg of mycelium was suspended in the solutions shown in the following table for 2 minutes and then filtered onto cotton wool and dried. After 3 days, the mycelium wafers were placed in petri dishes with a water-agar layer so that each petri dish had 25 large and 25 small (obtained by cutting large pieces into pieces) 10,57875 mycelium wafers. Petri dishes were examined after 21 days and the number of mycelium with growth of Rhizoctonian hyphae was calculated.

% number of live mycelium suspended or dissolved in water natarrycin ethyl alcohol acetic acid large small 0.2 50 0.1 0 0 0.2 10 0.1 0 0 0.2 0.1 3 7 untreated 23 23

The table shows that the combination of natamycin, ethyl alcohol and acetic acid is better than the combination of natamycin with acetic acid alone.

C. In another experiment, potatoes were immersed in the test solution compositions shown in the following table for 5 minutes at 12 ° C. The potatoes (Sientje) were then dried and allowed to germinate for 7 days at 25 ° C. The potatoes were then planted in glass cups on moist fine sand and kept at 14 ° C for 3 weeks. The moisture content of the sand was 20%.

After 3 weeks, each potato was examined for the presence of mycelium in the peel and germ and possible typical brown lesions caused by Rhizoctonia.

Some natamycin compositions, one crude crystalline natamycin and a product called 'kArdisar /' (based on ethyl mercuric bromide) were used as test disinfectants. There were 15 strongly infected potatoes in each experimental group. Potatoes with one damaged or mycelial sprout were considered disinfected.

11 57875

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The table shows that combinations with 0.2% natamycin in combination with alcohol and acetic acid have a disinfectant effect against Rhizoctonia solani that is at least comparable to standard treatment with a known composition. However, the standard treatment used in this experiment contains a mercury compound, and such compositions should preferably be avoided because of their toxicity. There is some evidence that propyl alcohol would be slightly better than ethyl alcohol.

Example 9 Iris bulbs, variety ‘’ Prof. Blaauw ", size 10. One experiment was performed in a greenhouse and the other outdoors.

For the greenhouse experiment, the bulbs were infected with a suspension containing 1.8 x 10 6 Fusarium oxysporium iris seed / ml.

One control group was left uninfected. Disinfection was performed in a 10 liter tank containing the disinfectant solutions shown in the following table. After disinfection, the bulbs were placed in small bags and air-dried at 17 ° C and 60% relative humidity for 11 days. After drying, the bulbs were planted in a greenhouse. The soil temperature was 18-21 ° C. Prior to flowering, the bulbs were examined for Fusarium infection.

Group% suspended or dissolved in water: natase streptomycin ethyl alcohol acetic acid "Benlat11 (1) infected onions A 0,04 0,008 - - - 42 B 0,04 0,008 20 0,1 - 8 C - - - 0,2 30 D - - - 309 E uninfected, non-disinfected 9 (1) water-miscible powder containing 50% methylbutylcarbamoylbenzimidazolylcarbamate.

Disinfectant solutions included some streptomycin as sulfate to avoid bacterial infections.

13 '57875

In the open-air experiments, the bulbs were treated in the same way, except that the inoculum suspension contained 1.5 x 10 disease seeds / ml. The temperature of the soil in which the bulbs were planted in the open air was 14-17 ° C. The results are shown in the table below.

Group% suspended or dissolved in water: natanyin streptonysin ΊΜΌ ethyl alcohol acetic acid "Benlate" infected onions (2) A 0,04 0,008 0,3 - - 17 B 0,04 0,008 0,3 20 0,1 - 4 C - -0 , 3 - - 0.2 10 D 0.3 168 E non-infected, non-disinfected 0 (2) water-miscible thiuramide sulfide composition added to avoid Pythium infection.

Both tables show that compositions B according to the invention are clearly superior to natamycin compositions without ethyl alcohol and acetic acid (compositions A) and that they are also better than "Benlate", which is another composition often used with flower bulbs.

Claims (7)

57875 li | The # numbers given in this patent application are generally volume / volume # 'and, except for the # number for natamycin or another polyene antibiotic, in which case it is weight / volume #. 1. An antifungal preparation for use in agricultural products such as fruits, vegetables, roots, beets, flower bulbs and rhizomes, characterized in that it contains about 0.04-0.2% of a polyene antibiotic containing natamycin, luzomomycin or aureofungin, dissolved to a mixture containing about 0.1-1% alkanoic acid having 1-8 carbon atoms, 1-3 carboxyl groups and 0-2 hydroxy groups, and from 10 # to the remainder of the mixture, preferably 15-50% alkanol having 1-3 carbon atoms. Preparation according to Claim 1, characterized in that the polyene antibiotic is natamycin or a salt or ester thereof. Preparation according to Claim 1 or 2, characterized in that the alkanol is ethyl alcohol. Preparation according to Claim 1, 2 or 3, characterized in that the alkanoic acid is acetic, propionic, lactic, citric or ascorbic acid. Preparation according to one of the preceding claims, characterized in that it further contains water and additives such as coating components, e.g. plastic emulsions, such as polyvinyl acetate emulsions, antioxidants, e.g. ascorbic acid, wetting agents and thickeners, such as gelatin. 1. Antifungals for the preparation and use of other products, as a whole, in the form of fruit, crumbs, crumbs, buttermilk and rotary grains, in the form of a product containing about 0.04 to 0.2 g of a polyanantibiotic, with the aid of natamycin , lucenzo-mycin or aureofungin, preferably in the range of about 0.1-1% with alkanes having 1-8 cholaters, 1-3 carboxyl groups and 0-2 hydroxyl groups as well as 10% up to the rest of the bleaching,