Vol. x(x), pp. xxxxx, x xx, 2016 DOI: xxxxxxxxxxxx Article Number: xxxxx ISSN 1684-5315 Copyright © 2016 Author(s) retain the copyright of this article http://www.academicjournals.org/AJB
African Journal of Biotechnology
Full Length Research Paper
Study of secondary metabolites produced by Aspergillus flavus and evaluation of the antibacterial and antifungal activity Ghaidaa J. Mohammed1, Imad H. Hameed2* and Sabreen A. Kamal2 1
College of Science, Al-Qadisia University, Iraq. Department of Biology, Babylon University, Iraq.
2
Received 25 October, 2015; Accepted 19 February, 2016
The objective of this study was the analysis of the secondary metabolite products and evaluation of antibacterial activity of Aspergillus flavus. Bioactives are chemical compounds often referred to as secondary metabolites. Thirty one bioactive compounds were identified in the methanolic extract of A. flavus. The identification of bioactive chemical compounds is based on the peak area, retention time, molecular weight and molecular formula. Gas chromatography mass spectrometry (GC-MS) analysis of A. flavus revealed the existence of the 1,2-cis-1,5-trans-2,5-dihydroxy-4-methyl-1-(1-htdroxy-1isopropyl)cy, 2-furancarboxaldehyde,5-methyl, 2(5H)-Furanone, 6-hydroxymethyl-5-methylbicyclo[3.1.0]hexan-2-one, D-glucose,6-O-α-D-galactopyranosyl, 2-(3-hydroxy-propyl)-cyclohexane-1,3dione, 9-oxa-bicyclo[3.3.1]nonane-1,4-diol, benzenemethanol,2-(2-aminopropoxy)-3-methyl, 1,2cyclopentanedione,3-methyl, α-D-glucopyranoside, O-α-D-glucopyranosyl-(1.fwdarw.3)-ß-D-fruc, 1-nitro2-acetamido-1,2-dideoxy-d-mannitol, desulphosinigrin, orcinol, bicyclo[2.2.1]heptane-2-carboxylic acid isobutyl-amide, 2H-oxecin-2-one.3.4.7.8.9.10-hexahydro-4-hydroxy-10-methyl-.[4, 2H-pyran,tetrahydro-2(12-pentadecynyloxy), maltol, 2-tridecyl-5-(acetylamino)tetrahydro-γ-pyrone, cycloundecanone , oxime, D-glucose,6-O-α-D-galactopyranosyl, 6-acetyl-ß-d-mannose, 5-hydroxymethylfurfural, 1-gala-l-idooctonic lactone, pterin-6-carboxylic acid, uric acid, acetamide , N-methyl -N-[4-[2-acetoxymethyl-1pyrrolidyl]-2-butynyl], l-(+)-ascorbic acid 2,6-dihexadecanoate, D-fructose , diethyl mercaptal , pentaacetate, 2-bromotetradecanoic acid, octadecanal ,2 –bromo, L-ascorbic acid, 6-octadecanoate, and 18,19-secoyohimban-19- oic acid,16,17,20,21-tetradehydro-16. The FTIR analysis of A. flavus proved the presence of aromatic rings, alkenes, aliphatic fluoro compounds, aromatic nitro compounds, ammonium ions and organic nitrate which shows major peaks at 667.37, 694.37, 904.61, 1028.06, 1145.43, 1205.51, 1234.44, 1273.02, 1311.94, 1377.17, 1413.82, 1450.47, 1492.90, 1600.92, 1629.99, 2852.72, 2922.16, 3024.38, 3271.27 and 3286.7. Datura stramonium was very highly antifungal activity (9.00±0.73) mm while Escherichia coli has maximum zone formation (7.55±0.09) mm against A. flavus. Key words: Aspergillus flavus, antibacterial activity, antifungal activity, Fourier transform infrared spectroscopy (FTIR), gas chromatography mass spectrometry (GC/MS), secondary metabolites. INTRODUCTION The genus Aspergillus belongs to the Deuteromycota
division of the fungi kingdom. The genus comprises
approximately 180 species, of which 33 have been associated with human disease (Segal et al., 1998; Perfect et al., 2001). A culture yielding Aspergillus spp., in addition to enabling a diagnosis of invasive aspergillosis, may further define therapeutic options via susceptibility testing or the isolation of a species possessing inherent antifungal resistance; examples of the latter include Aspergillus terreus and Aspergillus nidulans, which are both resistant to amphotericin B (Walsh, 2004; Susca et al., 2010; Al-Marzoqi et al., 2015). Some species of the genus produce secondary metabolites in food as aflatoxins (AFs) which are produced mainly by Aspergillus flavus and Aspergillus parasiticus. A. flavus has been associated with secondary respiratory infections in immuno-compromised human patients (Hedayati et al., 2007; Perrone et al., 2007; Mogensen et al., 2009; Altameme et al., 2015a; Hameed et al., 2015a). Aflatoxins are most toxic and most potent carcinogenic natural compounds that cause aflatoxicosis and induce cancers in mammals. A. flavus is also an opportunistic pathogen and has been isolated from insects, birds, mammals, and plants and widely distributed soil-borne molds and can be found anywhere on earth. It can reproduce abundantly resulting from the production of numerous airborne conidia. The spores can easily disperse by air. Environment has a great impact on mould growth, with humidity being the most important variable (McAlpin et al., 2002; Hedayati et al., 2007). It is a saprophytic fungus that is capable of surviving on many organic nutrient sources like plant debris, tree leaves, decaying wood, animal fodder, cotton, compost piles, dead insect and animal carcasses, outdoor and indoor air environment (air ventilation system), stored grains, and even human and animal patients (Hua et al., 2012). The aims of this study were the analysis of the secondary metabolites and evaluation of the antibacterial and antifungal activity. MATERIALS AND METHODS Growth conditions and determination of metabolites A. flavus was isolated from dried fruit and the pure colonies were selected, isolated and maintained in potato dextrose agar slants (Usha and Masilamani, 2013; Altameme et al., 2015b). Spores were grown in a liquid culture of potato dextrose broth (PDB) and incubated at 25°C in a shaker for 16 days at 130 rpm. The extraction was performed by adding 25 ml methanol to 100 ml liquid culture in an Erlenmeyer flask after the filtration of the culture. The mixture was incubated at 4°C for 10 min and then shook for 10 min at 130 rpm. Metabolites was separated from the liquid culture and evaporated to dryness with a rotary evaporator at 45°C. The residue was dissolved in 1 ml methanol, filtered through a 0.2 μm syringe filter, and stored at 4؛C for 24 h before being used for GC-
MS. The identification of the components was based on comparison of their mass spectra with those of NIST mass spectral library (Altameme et al., 2015b; Hameed et al., 2015b).
Analysis of bioactive compounds by chromatography-mass spectrometry (GC/MS)
using
gas
Bioactive compounds were examined for the chemical composition using GC-MS (Agilent 789A) equipped with a DB-5MS column (30 m × 0.25 mm i.d., 0.25 µm film thickness, J&W Scientific, Folsom, CA). The oven temperature was programmed as for the previous analysis. Helium was used as the carrier gas at the rate of 1.0 ml/min. Effluent of the GC column was introduced directly into the source of the MS via a transfer line (250°C). Ionization voltage was 70 eV and ion source temperature was 230°C (Hameed et al., 2016; Altameme et al., 2015c; Muhanned et al., 2015). Scan range was 41- 450 amu. The constituents were identified after compared with available data in the GC-MS library in the literatures (Hamza et al., 2015; Hameed et al., 2015c).
Fourier transform infrared spectrophotometer (FTIR) The powdered sample of the A. flavus specimen was treated for Fourier transform infrared spectroscopy (Shimadzu, IR Affinity 1, Japan). The sample was run at infrared region between 400 and 4000 nm (Mohammed and Imad, 2013; Idan et al., 2015).
Determination of antibacterial activity The test pathogens (Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, and E. coli) were swabbed in Muller Hinton agar plates. 90 μl of fungal extracts was loaded on the bored wells. The wells were bored in 0.5 cm in diameter. The plates were incubated at 37°C for 24 h and examined (Anupama et al., 2007). After the incubation the diameter of inhibition zones around the discs was measured.
Determination of antifungal activity A. flavus isolate was suspended in potato dextrose broth and diluted to approximately 105 colony forming unit (CFU) per ml. They were flood inoculated onto the surface of Potato dextrose agar and then dried. Standard agar well diffusion method was followed (Rajasekar et al., 2012; Tabaraie et al., 2012; Gebreselema et al., 2012; Usha and Masilamani, 2013). Five-millimeter diameter wells were cut from the agar using a sterile cork-borer, and 25 μl of the samples solutions (Linum usitatissimum, Anastatica hierochuntica, Gramineae poaceae, Nerium olender, Ricinus communis, and Datura stramonium) were delivered into the wells. The plates were incubated for 48 h at room temperature. Antimicrobial activity was evaluated by measuring the zone of inhibition against the test microorganisms. Methanol was used as solvent control. Amphotericin B and fluconazole were used as reference antifungal agent (Anesini and Perez, 1993; Rukayadi et al., 2006; Hameed et al., 2015d; Jasim et al., 2015). The tests were carried out in triplicate. The antifungal activity was evaluated by measuring the inhibition-zone diameter observed after 48 h of incubation (Bellini et al., 2003; Idan et al., 2015).
*Corresponding author. E-mail:
[email protected]. Tel: 009647716150716. Author(s) agree that this article remains permanently open access under the terms of the Creative Commons Attribution License 4.0 International License
Figure 1. Morphological characterization of A. flavus colony.
glucose,6-O-α-D-galactopyranosyl, 2-(3-hydroxy-propyl)cyclohexane-1,3-dione, 9-Oxa-bicyclo[3.3.1]nonane-1,4diol, benzenemethanol,2-(2-aminopropoxy)-3-methyl, 1,2cyclopentanedione,3-methyl, α-D-glucopyranoside, O-αD-glucopyranosyl-(1.fwdarw.3)-ك-D-fruc, 1-nitro-2acetamido-1,2-dideoxy-d-mannitol, desulphosinigrin, orcinol, bicyclo[2.2.1]heptane-2-carboxylic acid isobutylamide, 2H-oxecin-2-one.3.4.7.8.9.10-hexahydro-4hydroxy-10-methyl-.[4,2H-pyran,tetrahydro-2-(12pentadecynyloxy), maltol, 2-tridecyl-5(acetylamino)tetrahydro-γ-pyrone, cycloundecanone , oxime, D-glucose,6-O-α-D-galactopyranosyl, 6-acetyl-كd-mannose, 5-hydroxymethylfurfural, 1-gala-l-ido-octonic lactone, pterin-6-carboxylic acid, uric acid, acetamide , Nmethyl -N-[4-[2-acetoxymethyl-1-pyrrolidyl]-2-butynyl], l(+)-Ascorbic acid 2,6-dihexadecanoate, D-fructose , diethyl mercaptal , pentaacetate, 2-bromotetradecanoic acid, octadecanal ,2 –bromo, L-ascorbic acid , 6octadecanoate, 18,19-secoyohimban-19oic acid,16,17,20,21-tetradehydro-16 (Figures 5 to 35). Many compounds are identified in the present study. Some of them are biological compounds with antimicrobial activities.
Statistical analysis Data were analyzed using analysis of variance (ANOVA) and differences among the means were determined for significance at P < 0.05 using Duncan’s multiple range test (by SPSS software) Version 9.1.
RESULTS AND DISCUSSION Based on morphological characteristics, fungi was isolated in selective media of potato dextrose agar media. Morphological, microscopical and microscopical characteristics of fungal strains were determined using specific media light and compound microscope (Figure 1). The 400 ml of fermentation broth (PDA broth) which contain 200 μl of the standardized fugal suspensions were used to inoculate the flasks and incubated at 37°C on a shaker at 90 rpm for 7 days. After fermentation, the secondary metabolites were produced by isolated microorganisms.
Identify the secondary metabolites from A. flavus Gas chromatography and mass spectroscopy analysis of compounds was carried out in methanolic extract of A. flavus, shown in Table 1. The GC-MS chromatogram of the thirty one peaks of the compounds detected IS shown in Figure 2. The first set up peak were determined to be 1,2-cis-1,5-trans-2,5-dihydroxy-4-methyl-1-(1-htdroxy-1isopropyl)cy (Figure 3). The second peak indicated to be 2-furancarboxaldehyde,5-methyl (Figure 4). The next peaks considered to be 2(5H)-furanone, 6hydroxymethyl-5-methyl-bicyclo[3.1.0]hexan-2-one, D-
Identification of the secondary metabolites from A. flavus by Fourier-Transform Infrared analysis The FTIR analysis of A. flavus proved the presence of aromatic rings, alkenes, aliphatic fluoro compounds, aromatic nitro compounds, ammonium ions and organic nitrate which shows major peaks at 667.37, 694.37, 904.61, 1028.06, 1145.43, 1205.51, 1234.44, 1273.02, 1311.94, 1377.17, 1413.82, 1450.47, 1492.90, 1600.92, 1629.99, 2852.72, 2922.16, 3024.38, 3271.27 and 3286.7 (Table 2; Figure 36). Antibacterial and antifungal activity Clinical pathogens selected for antibacterial activity namely, (Staphylococcus aureus. Pseudomonas aeroginosa, Klebsiella pneumoniae, and E. coli, maximum zone formation against E. coli (7.55±0.09) mm, Table 3, Figure 37. In agar well diffusion method the selected medicinal plants (Nerium olender, Ricinus communis, Datura stramonium, Linum usitatissimum, Anastatica hierochuntica, and Gramineae poaceae) were effective against A. flavus (Figure 38 and Table 4). Datura stramonium was very highly active (9.00±0.73) mm against A. flavus. A. flavus was found to be sensitive to all test medicinal plants and mostly comparable to the standard reference antifungal drug Amphotericin B and fluconazole to some extent. Conclusion The results of this study showed that A. flavus species
Table 1. Major bioactive chemical compounds identified in methanolic extract of A. flavus.
S/N
Bioactive compound
RT (min)
Formula
Molecular weight
Exact mass
1.
1,2-cis-1,5-trans-2,5dihydroxy-4-methyl-1-(1htdroxy-1-isopropyl)cy
3.585
C10H18O3
186
186.125594
59,71,81,95,110,135,1 53,168
2.
2Furancarboxaldehyde,5methyl
3.613
C6H6O2
110
110.0367794
53,81,95,110
3.
2(5H)-Furanone
3.831
C4H4O2
84
84.021129
55,84
4.
6-Hydroxymethyl-5methylbicyclo[3.1.0]hexan-2-one
3.859
C8H12O2
140
140.08373
55,69,81,95,110,125
5.
D-Glucose,6-O-α-Dgalactopyranosyl
3.997
C12H22O11
342
342.11621
60,73,85,110,126,144, 164,182,212,261
Chemical structure
MS Fragment- ions
Table 1. Contd.
6.
2-(3-Hydroxy-propyl)cyclohexane-1,3-dione
4.408
C9H14O3
170
170.094295
55,70,84,112,124,152
7.
9-Oxabicyclo[3.3.1]nonane-1,4diol
4.466
C8H14O3
158
158.094295
58,70,86,97,112,130,1 40,158
8.
Benzenemethanol,2-(2aminopropoxy)-3-methyl
4.546
195
195.125929
58,77,91,121,152,178
9.
1,2-Cyclopentanedione,3methyl
4.712
C6H8O2
112
112.0524297
55,83,97,112
10.
α-D-Glucopyranoside, Oα-D-glucopyranosyl(1.fwdarw.3)-ß-D-fruc
4.820
C18H32O16
504
504.169035
60,73,85,97,113,126,1 45,163,181,199
Table 1. Contd.
11.
1-Nitro-2-acetamido-1,2dideoxy-d-mannitol
4.901
252
252.095751
60,73,86,98,114,130,1 61,188,219,252
12.
Desulphosinigrin
5.009
279
279.077658
60,73,85,103,127,145, 163,213,262
13.
Orcinol
5.175
C7H8O2
124
124.0524297
67,95,107,124
14.
Bicyclo[2.2.1]heptane-2carboxylic acid isobutylamide
5.284
C12H21NO
195
195.162314
57,67,79,95,128,140,1 54,166,180,195
Table 1. Contd.
15.
2H-Oxecin-2one.3.4.7.8.9.10hexahydro-4-hydroxy-10methyl-.[4
5.341
C10H16O3
184
184.109944
55,70,81,112,124,142, 166,184
16.
2H-Pyran,tetrahydro-2(12-pentadecynyloxy)
5.536
C20H36O2
308
308.27153
55,67,85,101,161,184, 208,255,279,307
17.
Maltol
5.616
C6H6O3
126
126.031694
55,71,80,97,111,126
18.
2-Tridecyl-5(acetylamino)tetrahydro-γpyrone
5.782
C20H37NO3
339
339.277344
55,72,85,99,114,128,1 86,210,237,250,281,30 9,321,339
19.
Cycloundecanone , oxime
5.890
C11H21NO
183
183.162314
55,73,82,95,124,140,1 51,166
20.
6-Acetyl-ß-d-mannose
6.245
C8H14O7
222
222.073953
60,73,81,97,109,126,1 44,173,192
Table 1. Contd.
21.
5-Hydroxymethylfurfural
7.149
C6H6O3
22.
1-Gala-l-ido-octonic lactone
8.660
C8H14O8
23.
Pterin-6-carboxylic acid
24.
25.
126
126.031694
53,69,81,97,126
238
238.068868
61,73,84,112,127,142, 159,189,220
8.820
207
207.039239
57,69,105,163,207
Uric acid
9.701
168
168.02834
54,69,82,91,97,125,14 0,168
Acetamide , N-methyl -N[4-[2-acetoxymethyl-1pyrrolidyl]-2-butynyl]-
14.908
266
266.163042
55,67,82,91,124,141,1 65,193,251
Table 1. Contd.
652
652.49142
57,73,85,98,115,129,1 43,157,185,199,213,22 7,256,297,322,353
15.349
496
496.14369
60,69,113,129,154,185 ,213,,245,273,316
2-Bromotetradecanoic acid
16.694
306
306.119442
55,73,83,99,111,138,1 53,201,227,249,306
29.
Octadecanal ,2 –bromo
16.860
346
346.187128
57,83,95,109,122,137, 166,224,249,281,304,3 46
30.
L-Ascorbic acid , 6octadecanoate
17.084
442
442.293055
57,69,85,97,111,129,1 43,171,185,199,227,24 1,255,267,284,327,352
31.
18,19-Secoyohimban-19oic acid,16,17,20,21tetradehydro-16
17.186
352
352.178692
57,69,85,95,111,126,1 49,167,221,256,279,29 3,352
26.
l-(+)-Ascorbic acid 2,6dihexadecanoate
15.183
27.
D-fructose , diethyl mercaptal , pentaacetate
28.
C38H68O8
C24H42O7
Figure 2. GC-MS chromatogram of methanolic extract of A. flavus.
Figure 3. Mass spectrum of 1,2-cis-1,5-trans-2,5-dihydroxy-4methyl-1-(1-htdroxy-1-isopropyl)cy with retention time (RT)= 3.585.
Figure 4. Mass spectrum of 2-Furancarboxaldehyde,5-methyl with retention time (RT)= 3.613.
Figure 5. Mass spectrum of 2(5H)-Furanone with retention time (RT)= 3.831.
Figure 7. Mass spectrum of D-Glucose,6-O-α-Dgalactopyranosyl with retention time (RT)= 3.997.
Figure 6. Mass spectrum of 6-Hydroxymethyl-5-methylbicyclo[3.1.0]hexan-2-one with retention time (RT)= 3.859.
Figure 8. Mass spectrum of 2-(3-Hydroxy-propyl)-cyclohexane1,3-dione with retention time (RT)= 4.408.
Figure 9. Mass spectrum of 9-Oxa-bicyclo[3.3.1]nonane-1,4-diol with retention time (RT)= 4.466.
Figure 10. Mass spectrum of Benzenemethanol,2-(2aminopropoxy)-3-methyl with Retention Time (RT)= 4.546.
Figure 11. Mass spectrum of 1,2-Cyclopentanedione,3-methyl with retention time (RT)= 4.712.
Figure 12. Mass spectrum of α-D-Glucopyranoside, O-α-Dglucopyranosyl-(1.fwdarw.3)-ß-D-fruc with retention time (RT)= 4.820.
Figure 13. Mass spectrum of 1-Nitro-2-acetamido-1,2-dideoxy-dmannitol with retention time (RT)= 4.901.
Figure 14. Mass spectrum of Desulphosinigrin with retention time (RT)= 5.009.
Figure 15. Mass spectrum of Orcinol with Retention Time (RT)= 5.175.
Figure 16. Mass spectrum of Bicyclo[2.2.1]heptane-2-carboxylic acid isobutyl-amide with retention time (RT)= 5.284.
Figure 17. Mass spectrum of 2H-Oxecin-2-one.3.4.7.8.9.10hexahydro-4-hydroxy-10-methyl-.[4with Retention Time (RT)= 5.341.
Figure 18. Mass spectrum of 2H-pyran, tetrahydro-2-(12pentadecynyloxy) with retention time (RT)= 5.536.
Figure 19. Mass spectrum of maltol with retention time (RT)= 5.616.
Figure 20: Mass spectrum of 2-tridecyl-5-(acetylamino)tetrahydroγ-pyrone with retention time (RT)= 5.782.
Figure 21. Mass spectrum of Cycloundecanone, oxime with Retention Time (RT)= 5.890.
Figure 22. Mass spectrum of D-glucose,6-O-α-Dgalactopyranosyl with retention time (RT)= 6.165.
Figure 23. Mass spectrum of 6-Acetyl-ß-d-mannose with retention time (RT)= 6.245.
Figure 24. Mass spectrum of 5-hydroxymethylfurfural with retention time (RT)= 7.149.
Figure 25. Mass spectrum of 1-gala-l-ido-octonic lactone with retention time (RT)= 8.660.
Figure 27. Mass spectrum of uric acid with retention time (RT)= 9.701.
Figure 26. Mass spectrum of Pterin-6-carboxylic acid with Retention Time (RT)= 8.820.
Figure 28. Mass spectrum of 1-gala-l-ido- octonic lactone with retention time (RT)= 11.704.
Figure 29. Mass spectrum of acetamide, N-methyl -N-[4-[2acetoxymethyl-1-pyrrolidyl]-2-butynyl] with retention time (RT)= 14.908.
Figure 30. Mass spectrum of l-(+)-ascorbic dihexadecanoate with retention time (RT)= 15.183.
acid
2,6-
Figure 31. Mass spectrum of D-fructose, diethyl mercaptal , pentaacetate with retention time (RT)= 15.349.
Figure 32. Mass spectrum of 2-bromotetradecanoic acid with retention time (RT)= 16.694.
Figure 33. Mass spectrum of octadecanal ,2 –bromo with retention time (RT)= 16.860.
Figure 34. Mass spectrum of L-Ascorbic acid , 6-octadecanoate with retention time (RT)= 17.048.
Figure 35. Mass spectrum of 18,19-Secoyohimban-19acid,16,17,20,21-tetradehydro-16with retention time (RT)= 17.186.
oic
Table 2. Fourier-transform infrared spectroscopy peak values of A. flavus.
S/N 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22.
Peak (Wave number cm-ˡ) 667.37 694.37 746.45 904.61 1028.06 1068.56 1145.43 1205.51 1234.44 1273.02 1311.94 1377.17 1413.82 1450.47 1492.90 1600.92 1629.99 2852.72 2922.16 3024.38 3271.27 3286.70
Intensity 65.830 44.119 66.019 77.899 60.696 63.577 75.195 80.251 79.990 80.114 78.582 74.443 76.239 70.075 74.585 76.732 74.056 80.017 72.767 85.338 81.757 81.815
Bond C-H C-H C-H C-F stretch C-F stretch C-F stretch C-H -CH3 O-H O-H
Functional group assignment Unknown Aromatic rings Aromatic rings Alkenes Aliphatic fluoro compounds Aliphatic fluoro compounds Aliphatic fluoro compounds Tetiary amine, C-N stretch Unknown Unknown Aromatic nitro compounds Aromatic nitro compounds Ammonium ions Methyl-CH. asym Unknown Unknown Organic nitrate Methylene-CH. asym Methylene-CH. asym Unknown Normal polymeric O-H stretch Normal polymeric O-H stretch
Group frequency 690-900 690-900 675-995 1000-10150 1000-10150 1000-10150 1150-1207 1310-1390 1310-1390 1390-1430 1430-1470 1620-1640 2840-2860 2915-2935 3200-3400 3200-3400
Figure 36. Fourier-transform infrared spectroscopy peak values of A. flavus.
Table 3. Zone of inhibition (mm) of test bacterial strains to Aspergillus flavus bioactive compounds and standard antibiotics.
Fungal / Products antibiotics Streptomycin Rifambin Kanamycin Cefotoxime Fungal products
Staphylococcus aureus 1.77±0.22 1.60±0.09 0.58±0.09 1.99±0.25 6.41±0.32
Bacteria Pseudomonas Klebsiella eurogenosa pneumonia 1.31±0.09 1.00±0.21 1.76±0.13 1.05±0.09 0.55±0.38 0.93±0.29 1.65±0.18 1.74±0.39 5.17±0.08 7.22±0.09
Proteus mirabilis 1.08±0.13 0.5±0.11 0.5±0.22 1.31±0.34 6.71±0.14
Escherichia coli 1.91±0.05 0.86±0.03 0.44±0.09 1.38±0.01 7.55±0.09
produce many important secondary metabolites with high biological activities. Based on the significance of employing bioactive compounds in pharmacy to produce drugs for the treatment of many diseases, the purification of compounds produced by A. flavus species can be useful.
Conflict of Interests The authors have not declared any conflict of interests.
ACKNOWLEDGEMENT
Figure 37. Antibacterial activity of A. flavus.
Author sincerely wish to thank Dr. Ali Al-Marzuqi for providing the opportunity to work on this project. Am thankful to you for helping me through the various analysis stages, and for providing helpful criticism and feedback throughout the writing process.
Figure 38. Antifungal activity of extract plant on A. flavus.
Table 4. Zone of inhibition (mm) of test different bioactive compounds and standard antibiotics of plants to A. flavus.
S/N 1. 2. 3. 4. 5. 6. 7. 8. 9.
Plant Linum usitatissimum(اCrude) Anastatica hierochuntica (Crude) Gramineae poaceae (Crude) Nerium olender (Alkaloids) Ricinus communis (Alkaloids) Datura stramonium(Alkaloids) Amphotericin B Fluconazol Control
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Zone of inhibition (mm) 5.00±0.90 4.18±0.12 8.32±0.09 6.21±0.17 3.62±0.19 9.00±0.73 6.10±0.31 8.04±0.21 0.00
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