Use of a bougainvillea extract for the preparation of a preparation for inhibiting air fungi

Abstract

The invention adopts representative varieties of Bougainvillea glabra, such as water red, Angus and small leaf purple, to carry out antibacterial experiments on Trichocladium, Cladosporium, Aspergillus flavus, Penicillium italicum and Trichoderma hamatum, and finds that the air fungi have good inhibition effect, which provides a new idea and method for the prevention and control of air fungi.

Description

Technical Field

[0001] The invention belongs to the field of antibacterial properties of natural medicines, and particularly relates to the inhibitory effect of water or alcohol extracts of bougainvillea on airborne fungi. Background Art

[0002] Botanical fungicides are a type of natural fungicide that utilizes antimicrobial substances contained in plants or induced plant defenses to kill or inhibit the growth and development of pathogens. These fungicides are made through extraction and processing techniques. Botanical fungicides offer four major advantages: 1) They are environmentally friendly, with active ingredients derived from natural substances that undergo smooth degradation pathways in nature after application, resulting in minimal environmental pollution. 2) They are less susceptible to developing drug resistance, as they contain a high concentration of fungicidal ingredients and have a unique mode of action, making it difficult for pathogens to develop resistance. 3) They are highly selective, with low toxicity to humans, livestock, and natural enemies, effectively protecting ecological balance while ensuring agricultural production. 4) They have diverse activity profiles. In addition to their fungicidal properties, some botanical fungicides also promote plant growth and enhance plant immunity, such as fungicides made from extracts of Psoralea corylifolia seeds, which exhibit broad-spectrum bactericidal activity, immune induction, and growth promotion.

[0003] Currently, the main means of preventing and controlling airborne fungi include environmental control, air purification, and chemical control. However, these methods each have their own shortcomings: environmental control has limited effects on old buildings and high-humidity climates, and is difficult to maintain long-term stability; HEPA and UV-C equipment have high operating costs and require regular replacement of filters or lamps, otherwise they are prone to failure; chemical disinfectants may leave toxic residues, causing secondary harm to humans and pets, and can cause fungi to develop drug resistance.

[0004] Therefore, it is urgent to find a new method to inhibit airborne fungi to reduce the impact on the environment and health. Summary of the Invention

[0005] The present invention provides a bougainvillea water extract or alcohol extract which can achieve an inhibitory effect only on airborne fungi, providing a new idea and method for inhibiting airborne fungi.

[0006] The invention provides use of a bougainvillea extract for preparing a preparation for inhibiting airborne fungi. The bougainvillea extract is a water extract or an alcohol extract.

[0007] The water extract of the present invention can be extracted by hot water extraction, cold water immersion, distillation extraction, etc.; the alcohol extract of the present invention can be extracted by hot alcohol extraction, cold alcohol immersion, reflux extraction, etc.

[0008] Furthermore, the airborne fungi include Trichoderma fragilis, Cladosporium cladosporium, Aspergillus flavus, Penicillium italicum and Trichoderma hooked.

[0009] Furthermore, the preparation also includes pharmaceutically acceptable excipients.

[0010] The present invention uses extracts of representative varieties of Bougainvillea, such as Water Red, Angus, and Small Leaf Purple Bracts and flowers, to carry out antibacterial experiments on Trichoderma fragilis, Cladosporium cladosporium, Aspergillus flavus, Penicillium italicum, and Trichoderma hooked, and finds that they have a good inhibitory effect on airborne fungi, providing a new idea and method for the prevention and control of airborne fungi. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 is the colony diameter of Mucor fragilis

[0012] Figure 2 Cladosporium cladosporum ( cladosporium cladosporioides ) colony diameter

[0013] Figure 3 Aspergillus flavus ( Aspergillus flavus ) colony diameter

[0014] Figure 4 Penicillium italicum ( penicillium italicum ) colony diameter

[0015] Figure 5 For Trichoderma hook trichoderma hamatum ) colony diameter

[0016] Figure 6 For Mucor fragilis ( Mucor fragiliss ) colony diameter

[0017] Figure 7 Cladosporium cladosporum ( Cladosporium cladosporioides ) colony diameter

[0018] Figure 8 For Trichoderma hook Trichoderma hamatum ) colony diameter

[0019] Figure 9 Fusarium spp. Fusarium proliferatum ) colony diameter

[0020] Figure 10 Alternaria alternata Alternaria alternata ) colony diameter

[0021] Figure 11 Fusarium solani ( Fusarium solani ) colony diameter

[0022] Figure 12 Fusarium oxysporum ( fusarium oxysporum ) colony diameter DETAILED DESCRIPTION

[0023] Example 1 Preparation of Bougainvillea water extract and alcohol extract

[0024] Experimental materials: bracts of three varieties of Bougainvillea: Water Red, Angus, and Small Leaf Purple. Flatten and dry in the shade for later use.

[0025] Collect bougainvillea bracts, air-dry the plants, and then dry them in a 28°C oven until crisp. Use a high-speed plant grinder (DWF-100) to grind them into a 60-mesh fine powder, weigh them, and set aside. Place 50g of the dry powder in a 1L beaker and add 500mL of distilled water. Soak overnight, then boil for 2 hours and filter. The filtrate is concentrated to 100mL in a flask to prepare a 0.5g / mL aqueous extract of bougainvillea stems. Place the extract in a sealed bag, label it, and store frozen.

[0026] Weigh 50 g of plant dry powder into a conical flask, add 250 mL of analytical grade ethanol, and perform ultrasonic-assisted extraction. The extract is filtered under reduced pressure and placed on a rotary evaporator (Shanghai Yarong Biochemical Instrument Factory). Concentrate to a paste under reduced pressure at 42°C and 247 MPa, transfer it to a brown wide-mouth bottle, weigh it, label it, and store it in a 4°C refrigerator for later use.

[0027] Example 2 Preparation of culture medium containing aqueous extract and alcoholic extract

[0028] After sterilizing the Bougainvillea aqueous extract, weigh 60mL and 120mL of each using a graduated cylinder in a clean bench. Cool the sterilized PDA medium to approximately 50°C, add the weighed aqueous extract, and mix thoroughly to achieve concentrations of 0.1g / mL and 0.2g / mL, respectively. Pour the diluted medium onto plates and let cool before use.

[0029] Remove the bougainvillea paste and weigh 6g of the paste into a small vial. Add 6mL of analytical grade ethanol and sonicate for 3 hours to thoroughly mix. Prepare a 1g / mL stock solution (①), label it, and set aside. Take 3mL of stock solution (①), add 3mL of analytical grade ethanol, mix thoroughly, and prepare a 0.5g / mL stock solution (②), label it, and set aside. Prepare and sterilize PDA culture medium. In a laminar flow hood, add stock solutions (①) and (②) to the PDA culture medium, bringing the ethanol extract concentrations to 0.01g / mL and 0.005g / mL, respectively. Pour the diluted culture medium into a plate and let it cool before use.

[0030] Example 3 Antibacterial Experiment

[0031] 1. Activation of pathogens

[0032] Under the aseptic operation of the clean bench, the target strain (Mucor fragilis ( Mucor fragilis ), Cladosporium cladosporium ( Cladosporium cladosporioides )、Aflatoxin ( Aspergillus flavus )、Penicillium italicum( Penicillium italicum )、Trichoderma hooked ( Trichoderma hamatum )) Inoculate onto a culture dish, culture upside down at 25℃ for 5-7 days, and set aside.

[0033] 2. Inoculation and measurement of colonies

[0034] Under aseptic conditions in a clean bench (Suzhou Antai Air Technology Co., Ltd.), the target bacterial strain was evenly punched along the edge of the bacterial colony with a microporator. A bacterial plate was picked with an inoculating needle and inoculated into the center of the culture dish. For strains with a high number of spores, two drops of 0.5% Tween reagent were added to emulsify the solution. 50 μL (or 1-2 drops) of the bacterial solution was pipetted from the tube where the solution had been dissolved and dripped into the center of the PDA culture medium. Each gradient was repeated three times.

[0035] Seal the inoculated culture dish and place it in an incubator, inverted, for 4-7 days. Observe the colonies on the dish and measure their diameters with a ruler. Enter these diameters into a table and calculate statistical data such as the mean diameter and standard deviation.

[0036] 3. Experimental results

[0037] 3.1 Experimental results of Bougainvillea water extract on target bacterial species

[0038] 3.1.1 Experimental results of culture medium

[0039] 5 days after inoculation, Trichoderma fragilis ( Mucor fragilis ) The average colony diameter in PDA medium reached 4.4 cm. The colony diameters in the medium containing 0.1 g / mL and 0.2 g / mL of the water extract of Red Globe were slightly smaller than those in the control group, at 3.8 cm and 3.6 cm, respectively. The colony diameters in the medium containing 0.1 g / mL and 0.2 g / mL of the water extract of Purple Globe were 3.8 cm and 3.0 cm, respectively; and the colony diameters in the medium containing 0.1 g / mL and 0.2 g / mL of the water extract of Angus were 3.1 cm and 2.0 cm, respectively. Figure 1 It can be seen that the water extract of Bougainvillea has a significant inhibitory effect on Trichoderma fragilis.

[0040] 5 days after inoculation, Cladosporium cladosporium ( cladosporium cladosporioides) The average diameter of the colonies in PDA medium reached 3.1333 cm. The colony diameters in the medium with the concentration of water red extract of 0.1g / mL and 0.2g / mL were smaller than those in the control group, which were 2.9333cm and 0.5 cm respectively; the colony diameters in the medium with the concentration of water purple extract of 0.1g / mL and 0.2g / mL were 2.3cm and 1.7cm respectively; the colony diameters in the medium with the concentration of Angus extract of 0.1g / mL and 0.2g / mL were 2.1cm and 2.3cm respectively. Figure 2 It can be seen that the water extract of Bougainvillea has a significant inhibitory effect on Alternaria alternata.

[0041] After 5 days of inoculation, Aspergillus flavus ( Aspergillus flavus ) The average diameter of the colonies in PDA medium reached 3.4 cm, and it did not grow in Angus water extract concentrations of 0.1g / mL and 0.2g / mL. Figure 3 It can be seen that the water extract of Bougainvillea has a significant inhibitory effect on aflatoxin.

[0042] 5 days after inoculation, Penicillium italicum ( penicillium italicum ) The average diameter of the colonies in PDA medium reached 3.9 cm, while the diameters of the colonies in the medium with the concentration of water red extract of 0.1g / mL and 0.2g / mL were smaller than those in the control group, at 3.8cm and 3.4cm respectively. Figure 4 It can be seen that the water extract of Bougainvillea has a significant inhibitory effect on Penicillium italicum.

[0043] After 5 days of inoculation, Trichoderma hookii ( trichoderma hamatum ) The average diameter of the colonies in PDA medium reached 5.3 cm. The diameters of the colonies in the medium with the concentration of water red extract of 0.1g / mL and 0.2g / mL were significantly smaller than those in the control group, at 5.0cm and 2.3cm respectively; the diameters of the colonies in the medium with the concentration of water purple extract of 0.1g / mL and 0.2g / mL were 4.9cm and 3.3cm respectively; the diameters of the colonies in the medium with the concentration of water Angus extract of 0.1g / mL and 0.2g / mL were 5.0cm and 3.4cm respectively. Figure 5 It can be seen that the water extract of Bougainvillea has a significant inhibitory effect on Trichoderma hooked.

[0044] 3.1.2 EC50 value (EC50 (effective concentration 50): the half-maximal inhibitory concentration, also known as the half-maximal effect concentration, is often used as an indicator of drug safety. For this test, it refers to the concentration of Bougainvillea extract that inhibits 50% of the test bacteria)

[0045] According to the obtained EC50 values, the inhibitory effects of the aqueous extract of Bougainvillea against the tested fungi were ranked from best to worst as follows: Cladosporium cladosporium > Trichoderma uncinate > Trichoderma fragilis > Penicillium italicum > Aspergillus flavus, as shown in Table 1 below, indicating that the aqueous extract of Bougainvillea had significant inhibitory effects on these five airborne fungi.

[0046] Table 1 EC50 values ​​of fungi under the water extract of Polygonum multiflorum

[0047]

[0048] According to the obtained EC50 values, the inhibitory effects of the water extract of Gies against the test fungi were ranked from best to worst as follows: Aspergillus flavus > Trichoderma fragilis > Trichoderma cladocerum > Cladosporium cladocerum, as shown in Table 2 below, indicating that the water extract of Bougainvillea had a significant inhibitory effect on these five air fungi.

[0049] Table 2 EC50 values ​​of fungi under Angus water extract

[0050]

[0051] From the above experimental results, it can be seen that the water extract of Bougainvillea has a significant inhibitory effect on five common airborne fungi.

[0052] 3.2 Experimental results of Bougainvillea alcohol extract on target bacterial species

[0053] 3.2.1 Experimental results of culture medium

[0054] After 5 days of inoculation, the average colony diameter of Trichoderma fragilis in PDA medium reached 4.9 cm, and the colony diameters in the medium with 0.005 g / mL and 0.1 g / mL of water red alcohol extract were 4.0 cm and 3.96 cm, respectively. Figure 6 It can be seen that the ethanol extract of Bougainvillea showed an inhibitory effect on Trichoderma fragilis.

[0055] Four days after inoculation, the average colony diameter of Cladosporium cladosporium in PDA medium reached 3.7 cm, while that in the dilutions of the three varieties of Bougainvillea ethanol extracts was 0.5 cm, indicating abnormal growth. Figure 7 It can be seen that the ethanol extract of Bougainvillea showed obvious inhibitory effect on Cladosporium cladosporium.

[0056] After 4 days of inoculation, the average diameter of the hooked Trichoderma colonies in PDA medium reached 8.5 cm. In the medium with the concentration of 0.005 g / mL and 0.1 g / mL of the alcohol extract of water red, the colony diameters were 2.0 cm and 1.7 cm respectively; in the medium with the concentration of 0.005 g / mL and 0.1 g / mL of the alcohol extract of small leaf purple, the colony diameters were 4.7 cm and 3.1 cm respectively; in the medium with the concentration of 0.005 g / mL and 0.1 g / mL of the alcohol extract of Angus, the colony diameters were 3.5 cm and 1.97 cm respectively. Figure 8 It can be seen that the ethanol extract of Bougainvillea showed obvious inhibitory effect on Trichoderma hookii.

[0057] 3.2.2 EC50 value (EC50 (effective concentration 50): the half-maximal inhibitory concentration, also known as the half-maximal effect concentration, is often used as an indicator of drug safety. For this test, it refers to the concentration of Bougainvillea extract that inhibits 50% of the test bacteria)

[0058] According to the obtained EC50 values, the inhibitory effects of the alcohol extract of Bougainvillea rutaecarpa on the tested fungi were ranked from best to worst as follows: Cladosporium cladosporium > Penicillium italicum > Trichoderma hookedum > Aspergillus flavus > Trichoderma fragilis, as shown in Table 3 below, indicating that the alcohol extract of Bougainvillea rutaecarpa had significant inhibitory effects on these five air fungi.

[0059] Table 3 EC50 values ​​of fungi under the influence of water red alcohol extract

[0060]

[0061] According to the obtained EC50 values, the inhibitory effects of Angus alcohol extract on the test fungi were ranked from best to worst as follows: Trichoderma fragilis > Cladosporium cladosporium > Aspergillus flavus > Trichoderma hooked > Penicillium italicum, as shown in Table 4 below, indicating that Bougainvillea alcohol extract had significant inhibitory effects on these five airborne fungi.

[0062] Table 4 EC50 values ​​of fungi under Angus ethanol extract

[0063]

[0064] From the above experimental results, it can be seen that the ethanol extract of Bougainvillea has a significant inhibitory effect on five common airborne fungi.

[0065] Comparative Example 1 Effect of Bougainvillea Water Extract on Plant Pathogens

[0066] The antibacterial effects of the water extracts of three varieties of Bougainvillea, namely, Water Red, Angus, and Small Leaf Purple, on common pathogens were statistically analyzed. The experimental steps in this experiment were consistent with those in Examples 1-3.

[0067] After 5 days of inoculation, Fusarium spp. Fusarium proliferatum ) The average diameter of the colonies in PDA medium reached 3.8667 cm. The diameters of the colonies in the medium with the concentration of water red extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 4.7667cm and 4.8667 cm respectively. The diameters of the colonies in the medium with the concentration of water purple extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 4.4667cm. The diameters of the colonies in the medium with the concentration of Angus extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 5.1667cm and 4.9333 cm respectively. Figure 9 It can be seen from the results that the water extract extracted from Bougainvillea has no inhibitory effect on Fusarium spp. and promotes the growth of Fusarium spp.

[0068] 5 days after inoculation, Alternaria alternata ( Alternaria alternata ) The average diameter of the colonies in PDA medium reached 4.0 cm. The diameters of the colonies in the medium containing 0.1g / mL and 0.2g / mL of water extract of Red Globe were slightly larger than those in the control group, at 4.0cm and 4.3cm respectively. The diameters of the colonies in the medium containing 0.1g / mL and 0.2g / mL of water extract of Purple Globe were slightly larger than those in the control group, at 4.5cm and 5cm respectively. The diameters of the colonies in the medium containing 0.1g / mL and 0.2g / mL of water extract of Angus were slightly larger than those in the control group, at 3.9cm and 4.2cm respectively. Figure 10 It can be seen that the water extract extracted from Bougainvillea has no inhibitory effect on Alternaria alternata and promotes the growth of Alternaria alternata in the two water extract concentration gradients.

[0069] 5 days after inoculation, Fusarium solani ( Fusarium solani ) The average diameter of the colonies in PDA medium reached 3.8667 cm. The diameters of the colonies in the medium with the concentration of water red extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 4.2667 cm and 3.9333 cm respectively; the diameters of the colonies in the medium with the concentration of water purple extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 4.2cm and 3.9cm respectively; the diameters of the colonies in the medium with the concentration of Angus extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 4.3 cm. Figure 11 It can be seen that the water extract extracted from Bougainvillea has no inhibitory effect on Fusarium solani and promotes the growth of Fusarium solani in the two water extract concentration gradients.

[0070] 5 days after inoculation, Fusarium oxysporum ( fusarium oxysporum) The average diameter of the colonies in PDA medium reached 4.2 cm. The diameters of the colonies in the medium with the concentration of water red extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 5.5cm and 5.6cm respectively; the diameters of the colonies in the medium with the concentration of water purple extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 5.0cm and 5.4cm respectively; the diameters of the colonies in the medium with the concentration of water Angus extract of 0.1g / mL and 0.2g / mL were slightly larger than those in the control group, at 5.4cm and 4.7cm respectively. Figure 12 It can be seen that the water extract extracted from Bougainvillea has no inhibitory effect on Fusarium oxysporum and promotes the growth of Fusarium oxysporum.

Claims

1. A use of a Bougainvillea bract extract for preparing a preparation for inhibiting airborne fungi, characterized in that: The bougainvillea extract is a water extract or an ethanol extract, and the air fungus is Trichoderma fragilis, Cladosporium cladosporium, Aspergillus flavus, Penicillium italicum or Trichoderma hooked.

2. The use according to claim 1, characterized in that: The preparation also includes pharmaceutically acceptable excipients.

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