Abstract
Control of Aspergillus flavus in corn using natural materials I would like to thank SfAM for awarding me a President’s Fund grant to attend the 9th European Conference on Fungal Genetics held in Edinburgh, UK on 5-8 April 2008. I presented my paper on the organisms that produce the mycotoxin, aflatoxin, especially Aspergillus flavus. This can infect important food and feed crops, including corn, before, during and after harvest. These fungi are normal soil-borne inhabitants in our environment, growing on both living and decaying plant matter. Mycotoxins include metabolic by-products produced by a number of different fungi. The importance of this study comes from the prevalence of Aspergillus flavus producing aflatoxin in corn, wheat and barley crops. It is possible to control contamination by preventing the presence of the fungus (Aspergillus flavus) and removing or affecting its capability to produce the toxin. This could be done by using natural materials and may lead to the production of toxinfree food and feed and increased safety from a health point of view whilst preserving grain food wealth. I studied the control of Aspergillus flavus using natural materials namely, the essential oil marjoram (Origanum majorana L.) and salt (sodium chloride) and their effect on fungal growth and the quantity of aflatoxin production. Effect of different concentrations of marjoram (Origanum majorana L.) essential oil and salt (sodium chloride) on Aspergillus flavus: One millilitre of Aspergillus flavus containing 5.0x104cfu/ml was added to 100 grams of corn (sterilized in a 250 ml conical flask) and distributed without the addition of marjoram or salt (control sample). Also, one millilitre of the same Aspergillus flavus with the addition of different volumes of marjoram essential oil (0.05, 0.1, 0.2, 0.3, 0.5 and 0.7 ml) and one ml of the same Aspergillus flavus with the addition of different weight of salt (0.2, 0.4, 0.6, 1.0, 1.5 and 2.0 grams) were each added to 100 grams of sterilized corn (placed in a 250 ml sterilized conical flask) and mixed. Sterilised distilled water was added to raise the moisture content to 18% and the mixture was shaken to make it homogenous. These were stored at 25OC for 21 days. Sampling to determine Aspergillus flavus count and aflatoxin concentration was made at intervals of 7 days until 21 days. Adding 0.05 ml of marjoram essential oil did not stop the growth of Aspergillus flavus during storage and up to 21 days, in inoculated corn. At the beginning of storage Aspergillus flavus count was 4.0x102 cfu/g and gradually increased on day 21 to reach 1.5x103cfu/g. The corresponding values in the control sample were 4x102 Aspergillus flavus count and 0 ppb aflatoxin production at the beginning of storage and 3.2x103 Aspergillus flavus and 150 ppb aflatoxin production at the end of storage (21 days). This indicates that the addition of 0.05 ml marjoram caused a reduction of Aspergillus flavus count of 62.5% at 21 days of storage. Raising the level of marjoram to 0.1 ml inhibited the growth of Aspergillus flavus and aflatoxin production completely at 7 days until the end of storage. These results are in agreement with those of Paster et al., (1995) who reported that the essential oil of marjoram has been shown to inhibit mycelial growth and aflatoxin production in Aspergillus parasites and Aspergillus flavus — both fungi which grow on stored grains. This observation was noted clearly with other higher marjoram levels. It can be observed from the effect of different salt concentrations on A. flavus count during storage of inoculated corn up to 21 days that the control sample gave a gradual increase in A. flavus number during storage from 0 to 21 days. It increased from 6x102 cfu/g to 5x103cfu/g at 0 and 21 days storage period respectively. The corresponding values, in inoculated corn sample treated with 0.2 grams of salt were 6x102cfu/g to 9x102cfu/g at the same periods of storage. Although the salt concentration (0.2 gram) was quite low it still produced a slight effect towards reducing the rate of growth of A. flavus during storage. This trend increased during storage, and was more obvious when 0.4 grams of salt was added. At this concentration the number of A. flavus at 0, 7, 14 and 21 days storage period became 6x102, 6.3x102, 6.6x102 and 7x102cfu/g, respectively. At 0.6% salt concentration growth inhibition reached 83.3% and aflatoxin production reached 50 ppb between 0 and 21 days while the control sample reached 180 ppb during the same period. Hence this concentration is not suitable for controlling A. flavus growth during storage. Raising salt concentration to 1 gram caused a complete inhibition of A. flavus growth and aflatoxin production during the whole experimented storage periods (7, 14, and 21 days). The same observation was noted when 1.5 and 2 grams of salt were used. Accordingly the minimum inhibitory concentration to prevent A. flavus growth in corn is 1 gram salt/100 gram corn and can be successfully used in the storage of corn up to 21 days. It is also suitable to prevent aflatoxin production. Moreover salt is traditionally used as a food, feed ingredient and food preservative, so the hazard would appear negligible. It is possible to conclude that using marjoram at concentration of 0.1 ml and above per 100g corn at 18% moisture inhibited the growth of Aspergillus flavus completely at 7 days and up to the end of storage also prevented aflatoxin production. Also it can be concluded that the minimum inhibitory concentration to prevent A. flavus growth in corn is 1 gram salt/100 gram corn. This concentration can be successfully used in the storage of corn up to 21 days. It is also suitable to prevent aflatoxin production.
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Claims 13
Claim · model-written summary
At 0.05 ml marjoram oil, Aspergillus flavus counts rose during 21 days of corn storage.
- Taxon
- corn
Zea mays subsp. mayse0 - Treatment
- 0.05 ml of marjoram essential oile0
- Interactor
- Aspergillus flavus
Aspergillus flavuse0pathogen - Outcome
- Aspergillus flavus counte0
- Direction
- increasede0
- Value
- 1.5x103cfu/ge0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Adding 0.05 ml of marjoram essential oil did not stop the growth of Aspergillus flavus during storage and up to 21 days, in inoculated corn. At the beginning of storage Aspergillus flavus count was 4.0x102 cfu/g and gradually increased on day 21 to reach 1.5x103cfu/g.”
Claim · model-written summary
The 0.05 ml marjoram treatment reduced Aspergillus flavus count by 62.5% at 21 days.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- 0.05 ml marjorame0
- Interactor
- Aspergillus flavusAspergillus flavuse0pathogen
- Outcome
- Aspergillus flavus counte0
- Direction
- reductione0
- Value
- 62.5%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“addition of 0.05 ml marjoram caused a reduction of Aspergillus flavus count of 62.5% at 21 days of storage”
t0“corn”
Claim · model-written summary
At 0.1 ml, marjoram completely inhibited Aspergillus flavus growth in stored corn.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- marjoram to 0.1 mle0
- Interactor
- Aspergillus flavusAspergillus flavuse0pathogen
- Outcome
- growth of Aspergillus flavuse0
- Direction
- inhibitede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Raising the level of marjoram to 0.1 ml inhibited the growth of Aspergillus flavus and aflatoxin production completely at 7 days until the end of storage.”
t0“corn”
Claim · model-written summary
At 0.1 ml, marjoram completely inhibited aflatoxin production in stored corn.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- marjoram to 0.1 mle0
- Interactor
- Aspergillus flavusAspergillus flavuse0pathogen
- Outcome
- aflatoxin productione0
- Direction
- inhibitede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Raising the level of marjoram to 0.1 ml inhibited the growth of Aspergillus flavus and aflatoxin production completely at 7 days until the end of storage.”
t0“corn”
Claim · model-written summary
A 0.2 gram salt treatment slightly reduced the rate of Aspergillus flavus growth.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- salt concentration (0.2 gram)e0
- Interactor
- A. flavusAspergillus flavuse0pathogen
- Outcome
- rate of growth of A. flavuse0
- Direction
- reducinge0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Although the salt concentration (0.2 gram) was quite low it still produced a slight effect towards reducing the rate of growth of A. flavus during storage.”
t0“corn”
Claim · model-written summary
At 0.4 grams salt, Aspergillus flavus count reached 7x102 cfu/g at day 21.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- 0.4 grams of salte0
- Interactor
- A. flavusAspergillus flavuse0pathogen
- Outcome
- number of A. flavuse0
- Direction
- becamee0
- Value
- 7x102cfu/ge0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“when 0.4 grams of salt was added. At this concentration the number of A. flavus at 0, 7, 14 and 21 days storage period became 6x102, 6.3x102, 6.6x102 and 7x102cfu/g, respectively.”
t0“corn”
Claim · model-written summary
At 0.6% salt, Aspergillus flavus growth inhibition reached 83.3%.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- 0.6% salt concentratione0
- Outcome
- growth inhibitione0
- Direction
- inhibitione0
- Value
- 83.3%e0
- Comparator
- control samplee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“At 0.6% salt concentration growth inhibition reached 83.3% and aflatoxin production reached 50 ppb between 0 and 21 days while the control sample reached 180 ppb during the same period.”
t0“corn”
Claim · model-written summary
At 0.6% salt, aflatoxin production reached 50 ppb versus 180 ppb in the control.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- 0.6% salt concentratione0
- Outcome
- aflatoxin productione0
- Direction
- reachede0
- Value
- 50 ppbe0
- Qualifier
- 180 ppbe0
- Comparator
- control samplee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“At 0.6% salt concentration growth inhibition reached 83.3% and aflatoxin production reached 50 ppb between 0 and 21 days while the control sample reached 180 ppb during the same period.”
t0“corn”
Claim · model-written summary
One gram salt completely inhibited Aspergillus flavus growth across storage periods.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- salt concentration to 1 grame0
- Interactor
- A. flavusAspergillus flavuse0pathogen
- Outcome
- A. flavus growthe0
- Direction
- inhibitione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Raising salt concentration to 1 gram caused a complete inhibition of A. flavus growth and aflatoxin production during the whole experimented storage periods (7, 14, and 21 days).”
t0“corn”
Claim · model-written summary
One gram salt completely inhibited aflatoxin production across storage periods.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- salt concentration to 1 grame0
- Outcome
- aflatoxin productione0
- Direction
- inhibitione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Raising salt concentration to 1 gram caused a complete inhibition of A. flavus growth and aflatoxin production during the whole experimented storage periods (7, 14, and 21 days).”
t0“corn”
Claim · model-written summary
Salt at 1.5 grams also completely inhibited Aspergillus flavus growth and aflatoxin production.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- 1.5e0
- Interactor
- A. flavusAspergillus flavuse1pathogen
- Outcome
- A. flavus growth and aflatoxin productione1
- Direction
- inhibitione1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The same observation was noted when 1.5 and 2 grams of salt were used.”
e1“Raising salt concentration to 1 gram caused a complete inhibition of A. flavus growth and aflatoxin production”
t0“corn”
Claim · model-written summary
Salt at 2 grams also completely inhibited Aspergillus flavus growth and aflatoxin production.
- Taxon
- cornZea mays subsp. mayst0
- Treatment
- 2 grams of salte0
- Interactor
- A. flavusAspergillus flavuse1pathogen
- Outcome
- A. flavus growth and aflatoxin productione1
- Direction
- inhibitione1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The same observation was noted when 1.5 and 2 grams of salt were used.”
e1“Raising salt concentration to 1 gram caused a complete inhibition of A. flavus growth and aflatoxin production”
t0“corn”
Claim · model-written summary
The authors identify 1 gram salt per 100 gram corn as the minimum inhibitory concentration for storage up to 21 days.
- Taxon
- cornZea mays subsp. mayse0
- Interactor
- A. flavusAspergillus flavuse0pathogen
- Outcome
- 1 gram salt/100 gram corne0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“minimum inhibitory concentration to prevent A. flavus growth in corn is 1 gram salt/100 gram corn and can be successfully used in the storage of corn up to 21 days”
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