Egyptian Journal of Natural Toxins, Vol. 2, 39-56 (2005).

DETECTION AND ESTIMATION OF AFLATOXIN B1

IN FEEDS AND ITS BIODEGRADATION BY
BACTERIA AND FUNGI
Rania M. Azab1; Wael M. Tawakkol2*; Abdel-Rahman M.
Hamad1; Mohamed K. Abou-Elmagd3; Hassan M. El-Agrab4
and Mohamed K. Refai5
1
Department of Mycology & 3Department of Biochemistry, Animal Health
Research Institute, Dokki; 2Department of Microbiology, Faculty of Pharmacy,
Cairo University; 4Department of Veterinary Hygiene and Management &
5
Department of Microbiology, Faculty of Veterinary Medicine, Cairo University
Received: 13/10/2004 Accepted: 17/03/2005

ABSTRACT

Mycological examination of 500 samples of animal feeds and feed

ingredients for Aspergillus flavus and Aspergillus parasiticus revealed
that 180 (36%) samples yielded isolates of Aspergillus flavus and 65 (13%)
samples gave isolates of Aspergillus parasiticus. Testing of the same
samples for AFB1 contamination showed that 99 samples (19.8%) contained
AFB1 at a rate of 125 ppb, 45 samples (9%) at a rate of 25-50 ppb, 32
samples (6.4%) at a rate of 101-200 ppb and 19 samples (3.8%) contained
aflatoxin B1 at a rate of 201-2000 ppb. Screening of isolated strains of
Aspergillus flavus and Aspergillus parasiticus for aflatoxin B1 production by
culturing on YES medium supplemented with 0.019% P-cresol revealed that
81 (45%) out of 180 isolates of Aspergillus flavus and 16 (24.62%) out of
65 isolates of Aspergillus parasiticus produced aflatoxin B1. Testing the
ability of 4 Lactobacillus strains for removal of aflatoxin B1 from liquid
media after physical and chemical treatments revealed that the acidic and
heat treatments of bacterial pellets significantly enhanced their ability to
bind aflatoxin B1 but heat treatment was not as effective as acidic treatment.
Screening the ability of either intact mycelium or fragmented mycelium or
culture cell - free system of non - aflatoxin B1 producing Aspergillus flavus
and Aspergillus parasiticus indicated that fragmentation increased the
ability of tested strain to degrade aflatoxin B1. Culture cell free system
showed the highest percent of aflatoxin B1 degradation. Aspergillus flavus
showed higher percent of degradation than Aspergillus parasiticus.

Key words: Aflatoxin B1, Aspergillus flavus, Aspergillus parasiticus, Biodegradation

* Corresponding author: e-mail: wtawakkol@hotmail.com

40 Detection and Estimation of Aflatoxin B1 in Feeds and its
Biodegradation By Bacteria and Fungi

INTRODUCTION
The problem of fungal contamination of foods and feeds in Egypt has
been already discussed by Refai et al. (1968), Refai and El-Bahay (1968),
Refai and Sadek (1968), Refai and Loot (1970) and Refai et al. (1990). The
mould contamination, particularly with mycotoxin-producing fungi is a
world problem. Approximately 25% of the world crops are affected by
mycotoxins annually. This causes significant economic losses due to loss of
crops and animals. The aflatoxins are of great concern because of their
determintal effects on the health of domestic animals and humans
(Anonymous, 1989) as well as the economic loss resulting from the
condemnation of contaminated products. Experimental data gathered during
the last 3 decades on the loss of productivity in farm animals consuming
contaminated feeds and the carcinogenicity in experimental animals provide
sufficient evidence regarding the hazardous nature of aflatoxins (Palmgren
and Hayes, 1987). The positive correlation between the consumption of
aflatoxin-contaminated foods and the increase of the incidence of liver
cancer in several populations in South East Asia and Africa suggests the
threat posed to human health by aflatoxin (Peers and Linsell, 1973).
Aflatoxins are unique in being resistant to degradation under normal
food processing conditions (Ciegler and Vesonder, 1983). This makes the
selection of proper degradation methods that will effectively decompose
aflatoxins, while retaining the nutritive quality and palatability of the treated
food, a continuous challenge. Preventing the contamination of food by the
toxigenic fungi, Aspergillus flavus and Aspergillus parasiticus, is the most
rational and economic approach to avoid the potential hazards. However,
prevention is not always possible under certain agronomic and storage
practices. Therefore, decontamination has gained importance in salvaging
food already contaminated with toxic fungal metabolites. Detoxification of
food containing aflatoxins is a problem of current concern. It can be
accomplished by a variety of methods. Segregation of contaminated and
non-contaminated products is sometimes carried out by hand sorting. While
this method is effective it is very labor intensive and may only be
economical, where labor costs are not expensive (Brackett, 1987). Physical
approaches to aflatoxins destruction generally involve treating with heat,
UV, light or ionizing radiation (Aziz and Refai, 1989; Aziz et al., 1990;
Refai et al., 1995 a,b and 2003). Chemical degradation of aflatoxins is
usually carried out by chlorination, oxidizing or hydrolytic agents. Of these
methods ammoniation is the most widely accepted, however, effective
ammoniation can require expensive equipment and may result in losses in
 Azab, R.M. et al. 41

nutritional quality of the treated feed (Samarajeewa et al., 1990; Refai et al.,
1995a).
Many microorganisms including bacteria, yeasts, moulds, actinomycetes
and algae are able to remove or degrade aflatoxins in foods and feeds (Line
and Brackett, 1995). Some strains of lactic acid bacteria have been reported
to be effective in removing aflatoxin B1 from contaminated liquid media
(El-Nezami et al., 1998). Nakazato et al. (1990) found that four fungal
strains namely Aspergillus niger, Eurotium herbarioum, Rhizopus sp. and
non-aflatoxin producing Aspergillus flavus could convert aflatoxin B1 to
aflatoxicol and also convert AFL to AFB1.
So, this preliminary study was carried out in order to determine the level
of aflatoxin B1 contamination in animal feeds, to isolate Aspergillus flavus
and Aspergillus parasiticus from the examined feed samples, to test the
toxigenicity of the isolated strains of Aspergillus flavus and Aspergillus
parasiticus, to artificially produce aflatoxin B1 on semisynthetic media by
standard toxigenic strains of Aspergillus flavus and to investigate the ability
of dairy strains of lactic acid bacteria and also some fungal strains to
degrade aflatoxin B1.

MATERIALS AND METHODS

A total of five hundred samples of different feedstuffs received by the
Animal Health Research Institute to detect aflatoxin contamination was used
in this study. These were poultry ration, concentrates, processed animal
feeds, corn, rice, barely, soya bean, wheat bran, rice straw and hay.
The media used were Sabouraud’s dextrose agar medium (Cruickshank
et al., 1975), Czapek-Dox agar (Raper and Fennel, 1965), malt extract agar
(Raper and Fennel, 1965), potato-dextrose agar medium (Shotwell et al.,
1966), yeast extract sucrose medium (Bauer et al., 1983), de Man-Rogosa-
Sharpe (MRs) broth (Oxoid, UK), glucose-salts medium (Doyle and Marth,
1978) and aflatoxin salts medium (Doyle and Marth, 1978).
Standard aflatoxins B1, B2, G1 and G2 were purchased from Sigma
(USA). Toxigenic strain of Aspergillus flavus (ATCC 28542) was purchased
from M.R.C. (Microbial Research Center, Faculty of Agriculture, Ain
Shams University, Cairo, Egypt).
Lactobacillus casi, L. bulgaricus, L. halvaticus were obtained from Ch.
Hansen’s Laboratories, Copenhagen, Denmark.
Isolation of moulds from feeds was carried out as described by Refai
(1979). Suspected Aspergillus isolates were identified into species according
42 Detection and Estimation of Aflatoxin B1 in Feeds and its
Biodegradation By Bacteria and Fungi

to Raper and Fennel (1965). The isolates were subcultured onto malt extract
agar and Czapek-Dox agar then incubated at 25 °C for 10 days.
Identification of the colonies was carried out by careful observation of the
macroscopic and microscopic characteristics of the mould colonies.
Screening of isolated strains of Aspergillus flavus and Aspergillus
parasiticus for aflatoxin B1 production was carried out according to Bauer et
al. (1983), while extraction of aflatoxin B1 from feed samples was done
according to Roberts and Patterson (1975) and production of aflatoxin by
standard toxigenic strain of A. flavus was carried out according to Bauer et
al. (1983). Qualitative determination of aflatoxin was achieved by thin layer
chromatography (T.L.C.), (Anonymous, 1975), while quantitative
estimation of aflatoxin B1 by a fluorometeric method according to
Anonymous (1990).
Bacterial biotransformation of aflatoxin B1 was carried out according to
Haskard et al. (2001). All strains of lactobacilli were cultured for 24 h in de
Man Rogosa sharpe (MRS) broth under aerobic conditions at 37°C. The
suspensions were centrifuged at 1.700 X g for 15 minutes. The supernatant
was discarded and the bacterial pellets were washed twice with phosphate
buffered saline (PBS; pH 7.3, 0.01 M) and the concentration was adjusted to
2 X 1010 bacteria per 4 ml PBS (per tube). The bacterial pellets were divided
into 5 parts, which were subjected to different treatments: the first part was
treated by heat (boiled in 4 ml PBS for 1 hour), the second part by acid
(incubated in 4 ml of 2 M HCl for 1 hour), the third part by ethanol
(incubated in 4 ml of 70% ethanol for 1 hour) the fourth part by alkaline
(incubated in 4 ml of 1 N NaOH for 1 hour), while the fifth part was used as
control and it was incubated in 4 ml PBS for 1 hour. Acid, alkali and
ethanol treated bacterial pellets were then washed twice with 4 ml PBS then
centrifuged. The treated bacterial pellets were then re-suspended in 1.5 ml
of PBS containing 5 µg of AFB1, incubated for 4 hours at 37C and the
supernatants were analyzed for AFB1 residues by fluorometer. All assays
were performed in triplicate.
Fungal biodegradation of aflatoxin B1 was done according to Doyle and
Marth (1978).
Data were statistically analysed using analysis of variance (ANOVA)
and comparing between groups were performed using least significant
difference (LSD) and Duncan Multiple Range test for comparative of mean
at P < 0.05. Pearson correlation was performed at P < 0.05 and P < 0.01
according to Petrie and Watson (1999) and computerized using SPSS 11
(Anonymous, 2001).
 Azab, R.M. et al. 43

RESULTS
Incidence of Aspergillus flavus and Aspergillus parasiticus in the
examined feed samples:
The results in table (1) show that 180 (36%) feed samples yielded
isolates of A. flavus and 65 (13%) samples gave isolates of A. parasiticus.
The highest percentage of recovery was observed in corn (60%) for A.
flavus and in ration concentrate (14.85%) for A. parasiticus.

The incidence of aflatoxin B1 in the examined feed samples:

Table (2) shows the incidence of aflatoxin B1 in contaminated samples,
where 237 out of the 500 feed samples (47.4%) were positive for aflatoxin
B1 contamination, of which 99 samples (19.8%) contained aflatoxin B1 at a
rate of 1-25 ppb, 45 samples (9%) 26-50 ppb, 32 samples (6.4%) 101-200
ppb and 19 samples (3.8%) contained aflatoxin B1 at a rate of 201-2000 ppb.
The remaining 263 samples (52.6%) were negative for aflatoxin B1.

Aflatoxin B1 production by standard toxigenic strain of Aspergillus

flavus:
As shown in figure (1), the yeast extract sucrose (YES) medium
supplemented with sorbic acid or P. cresol was a good medium for
production of aflatoxin B1 by standard toxigenic strain of A. flavus. The
produced aflatoxin B1 started to be detectable only after 10 days of
incubation. The level was very low at this time but increased significantly
on the 14th day. The maximum amount of produced toxins was obtained
after 20 days. It is obvious from the results that the addition of sorbic acid,
even at low concentration affected the production of aflatoxin B1 seriously.
This becomes clear, if it is compared with the medium containing P. cresol
which revealed a production rate starting at the 17th day (223.7 ppb) and
reached the maximum on the 20th day (287.5 ppb).

Aflatoxin B1 production by isolated strains of Aspergillus flavus and

Aspergillus parasiticus:
As seen in table (3), 180 isolates of Aspergillus flavus and 65 isolates of
Aspergillus parasitcus were tested for aflatoxin B1 production by culturing
on YES medium supplemented with 0.019% P. cresol. In case of
Aspergillus flavus 81 (45%) out of 180 tested isolates were found to be
aflatoxin B1 producer, while in case of Aspergillus parasitcus 16 (24.62%)
44 Detection and Estimation of Aflatoxin B1 in Feeds and its
Biodegradation By Bacteria and Fungi

out of 65 tested isolates produced aflatoxin B1. More than one half of the
tested strains could not produce AFB1 in the medium (55% A. flavus and
75.4% A. parasitcus). On the other hand, 35% of A. flavus produced
amounts of AFB1 that ranged from 1-100 µg/ 25 ml YES; in comparison to
16.9% of A. parasitcus. Only less then 2% of both species could produce
more than 200 µg AFB1/ 25 ml YES.

Surface binding of aflatoxin B1 by Lactobacillus strains:

It is clear from table (4) that all strains of lactobacilli had the ability to
bind AFB1, however, they varied in the rate of binding. L. acidophilus was
the most active followed by L. casi, L. halvaticus and L. bulgaricus. This
binding capacity was slightly reduced when the bacterial pellets were treated
with alkaline or ethanol. On the other hand, the best enhancer of binding
was the acid treatment, followed by heating. The latter was most obvious in
case of L. acidophilus which showed the highest rate of binding among all
tested bacterial stains.

Fungal biodegradation of aflatoxin B1:

The percentage of aflatoxin B1 degradation differed with the age of
mycelium for both A. flavus and A. parasiticus. The highest rate of aflatoxin
B1 degradation was associated with 9 days old mycelia for both strains. The
degraded percentage of AFB1 in case of A. flavus was non- significantly
higher than that of A. parasticus. The percentage of aflatoxin B1 degradation
was apparently increased with fragmentation of mycelium for both strains.
The non-heated cell free-system showed higher percentage of aflatoxin B1
than that of fragmented mycelium. On the other hand, the heated cell-free
system showed zero percentage of aflatoxin B1 degradation (Figs. 2, 3 & 4).

DISCUSSION
Mycotoxins are considered unavoidable contaminants in foods and feed
stuffs because agronomical technology has not yet advanced to stage at
which preharvest infection of susceptible crops by fungi can be eliminated
(Clark et al., 1980). Aflatoxin B1 received greater attention than any other
mycotoxins because of its demonstrable carcinogenic effect in susceptible
animals and its acute toxic effects in human (Bressac et al., 1991).
Mycological examination of feeds revealed the recovery of 180 isolates
(36%) of Aspergillus flavus and 65 isolates (13%) of Aspergillus parasiticus
from the examined samples. Connole et al. (1981) reported that 200 isolates
 Azab, R.M. et al. 45

(56.33%) of Aspergillus flavus were recovered from 355 animal feed

samples, while Youssef et al. (1985) found that one sample out of 10
samples of corn and wheat was contaminated with aflatoxigenic strains of
Aspergillus flavus and Aspergillus parasiticus.
Results obtained in this study demonstrate the existence and distribution
of aflatoxin B1 in different examined feeds and feed ingredients, where
47.4% of the examined samples contained aflatoxin B1. The content of
aflatoxin B1 varied from 1 to 2000 ppb. Such high aflatoxin content is
alarming, as the maximal level allowed by the Food and Drug
Administration (FDA) is 20 ppb for all feeds and foods and 0.5 ppb for fluid
milk (Schuller et al., 1983). Similar results were reported by Girgis et al.
(1977) and Badria et al. (1993), who detected aflatoxin B1 at different
percentage in examined samples of feeds and feed ingredients at levels
above or below the permissible limit. This slight variation in percentages of
aflatoxin B1 contaminating samples may be due to differences in locality
from which samples were collected or differences in storage condition.
Screening of obtained isolates of A. flavus and A. parasiticus for
aflatoxin B1 production revealed that 81 (45%) out of 180 tested isolates
were found to be aflatoxin B1 producers, while in case of A. parasiticus 16
(24.62%) out of 65 tested isolates were found to be aflatoxin B1 producer.
This was in agreement with those reported by Diener and Davis (1968), who
mentioned that 60% of isolates of A. flavus group produced some aflatoxins.
On the other hand, Hassan (1990) screened 55 isolates of A. flavus and A.
parasiticus isolated from corn in Egypt. Forty isolates of A. flavus produced
both aflatoxin B1 and aflatoxin B2, while A. parasiticus isolates were non-
toxigenic.
The results of surface binding of aflatoxin B1 by Lactobacillus strain
revealed that physical and chemical treatments would induce marked
alteration in the removal of aflatoxin B1 by tested strains. The results have
clearly indicated that bacterial viability was not a prerequisite to aflatoxin
B1 binding. It was reported that the binding of aflatoxin B1 to lactic acid
bacteria is a physical phenomenon instead of metabolic degradation reaction
that is associated with the cell wall (Thyagaraja and Hosono, 1994). On the
other hand, Tanabe et al. (1991) suggested that such binding may take place
through binding to cell wall peptidoglycan. These two possibilities are
greatly affected by the type of treatments particularly acid and heat
treatments which may alter the membrane bilayers, exposing their inner
faces, reducing cross link and/ or increasing pore size.
46 Detection and Estimation of Aflatoxin B1 in Feeds and its
Biodegradation By Bacteria and Fungi

Acid treatment was the most effective, significantly increasing the

ability of tested isolates to remove more aflatoxin B1. These results agree
with that reported by El-Nezami et al. (1998), who found that HCl treatment
of L. rhamnosus GG and L. rhamnosus 705 pellets significantly enhanced
the binding ability of it toward aflatoxin B1. Also, Haskard et al. (2001)
observed that specific lactic acid bacterial strains removed aflatoxin B1
from liquid medium by physical binding. The acid treated bacterial pellets
retained the highest amount of aflatoxin B1. Binding of aflatoxin B1 was
suggested to be intracellular in case of acid treatments which allow more
stable AFB1-bacterial complex.
Similarly, heat treated bacteria removed more aflatoxin B1 than non-
heated bacteria although heat treatment was not as effective as HCl
treatment. Such finding has also been reported by Thyagaraja and Hosono
(1994). Moreover, El-Nezami et al. (1998) revealed that heat treatment of
bacterial pellets of Lactobacillus strains by autoclaving or boiling at 100C
in water bath significantly enhanced its binding ability to aflatoxin B1 but
not effective as acid treatment. Similar observation was also noticed by
Pierides et al. (2000) and Haskard et al. (2001), while Line and Brackett
(1995) found that heat treatment or inactivation of Flavobacterium
aurantiacum made it unable to remove aflatoxin B1 from phosphate buffer.
This disagreement may be due to difference in type of bacteria as the
mechanism of aflatoxin B1 removal in case of Flavobacterium aurantiacum
is enzymatic.
The result of fungal biodegradation revealed that the percentage of
aflatoxin B1 degradation differed with the age of mycelium for both A.
flavus and A. parasiticus. The highest rate of aflatoxin B1 degradation was
associated with nine days old mycelia for both fungi. The percentage of
aflatoxin B1 degradation significantly increased with fragmentation of
mycelium for both fungi. The non-heated-cell-free system showed higher
percentage of aflatoxin B1 degradation than that of fragmented mycelium.
On the other hand, the heated cell-free system showed zero percentage of
aflatoxin B1 degradation. These finding indicate that fragmentation of
mycelium is essential for biodegradation of aflatoxin B1. So fragmentation
was suggested to release intracellular factors, which are responsible for
biodegradation of aflatoxin B1 as suggested by Doyle and Marth (1978). On
the other hand, Nakazato et al. (1990) found that non-aflatoxin producing A.
flavus convert aflatoxin B1 to aflatoxicol and also convert AFL to AFB1.
These conversion were distinctly observed in cell free system obtained from
disrupted mycelia of A. flavus but not observed in culture filtrates from
intact mycelia (non- disrupted) of the same isolates and not observed when
 Azab, R.M. et al. 47

the cell free system was preheated at 100°C. These finding strongly
suggested that the inter conversion of AFB1 and AFL is mediated by intra
cellular enzymes of A. flavus.
Finally, it can be concluded that the study revealed a high incidence of
aflatxoin contaminated feed and feed ingredients and that bacterial removal
of AFB1 from food and feed may be used on large scale to minimize
economic loss due aflatoxin contamination and to improve animal health
condition but this will involve additional processing steps.

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428.
Tanabe, T.; Otani, H. and Hosono, A. (1991): Binding of mutagens with
cell wall peptidoglycan of Leuconostoc mesenteroides sub sp.
dextranicum T- 180. Milchwissenschaft, 46(4): 622-625.
Thyagaraja, N. and Hosono, A. (1994): Binding properties of lactic acid
bacteria from "Idly" towards food-borne mutagens. Food Chem.
Toxicol., 32: 805-809.
Youssef, N.; Refai, M.K.; Molla, A.H.; Kandil, S. and Gammal, H.F.
(1985): Mycotic flora and aflatoxin contamination of grains. J. Egypt.
Vet. Med. Ass., 45(4): 69-81.
 Azab, R.M. et al. 51

Table (1): The incidence of Aspergillus flavus and Aspergillus parasiticus

in the examined feed samples
No. of Aspergillus
Aspergillus flavus
Type of sample examined parasiticus
samples Number % Number %
Poultry ration 150 57 38.0 % 21 14.0 %
Corn 25 15 60.0 % 3 12.0 %
Ration concentrate 175 51 29.1 % 26 14.8 %
Processed animal feed 50 19 38.0 % 5 10.0 %
Rice straw and hay 25 7 28.0 % 3 12.0 %
Barley 25 12 48.0 % 2 8.0 %
Soya bean 25 11 44.0 % 2 8.0 %
Wheat bran 25 8 32.0 % 3 12.0 %
Total 500 180 36.0 % 65 13.0 %
52 Detection and Estimation of Aflatoxin B1 in Feeds and its
Biodegradation By Bacteria and Fungi
 Azab, R.M. et al. 53

Table (3): Aflatoxin B1 production by isolated strains of Aspergillus

flavus and Aspergillus parasiticus.

Zero µg Amount of AFB1/25 ml YES broth

No. of AFB1/25
Type of
tested ml YES
isolate 1- 100 µg 101- 200 > 200 µg
isolates broth
AFB1 µg AFB1 AFB1

No. % No. % No. % No. %

Aspergillus
180 99 55 63 35 15 8.3 3 1.7
flavus
Aspergillus
65 49 75.4 11 16.9 4 6.2 1 1.5
parasiticus

Table (4): The effect of physical and chemical treatments of

lactobacillus strains on their ability to bind aflatoxin B1.

Pre- % AFB1 removed

F-
treatment of L. L. casi L. halvaticus L. bulgaricus
calculated
lactobacilli acidophilus
Non 56.6 ± 1.19 22.40 ± 0.47 17.80 ± 0.56 16.30 ± 1.25 abD 139.265*
AD aBD abD
Heat 71.90 ± 2.26 41.80 ± 0.50 28.50 ± 2.65 33.50 ± 0.95 152.127*
AdE aBdE abdE abdE
Ethanol 46.50 ± 1.99 21.80 ± 0.51 18.00 ± 0.86 15.90 ± 0.32 329.557*
AdeF aBeF abeF abeF
Acid 87.00 ± 0.97 43.10 ± 0.72 56.30 ± 1.02 58.60 ± 1.90 365.718*
AdefG aBdefG abdefG abdefG
Alkaline 27.40 ± 1.13 12.00 ± 0.46 9.10 ± 0.30 8.30 ± 0.51 173.526*
Adefg aBdefg abCdefg abcdefg
F-calculated 394.04* 31.129* 185.108* 73.28* --

* Significantly different using ANOVA test at P < 0.05, Aa, Bb, Cc, Dd, Ee, Ff, Gg
Significantly different between two comparison groups against capital liter using LSD
at P < 0.05
 54 Detection and Estimation of Aflatoxin B1 in Feeds and its
Biodegradation By Bacteria and Fungi

350

300
Aflatoxin concentartion (ug) .

250

200 Control
0.025 sorbic acid
150 0.019 % P. cresol

100

50

0
3 7 10 14 17 20 23
Time of incubation (days)

Figure (1): Aflatoxin B1 production in yeast extract sucrose YES

medium supplemented with a specific concentration of antimicrobial
agents during 23 days incubation at room temperature

4.5
4

3.5
% of degraded AFB1

3
A. flavus
2.5
A. parasticus
2

1.5
1

0.5
0
5 6 7 8 9 10 11 12 13 14 15 16 17 18
Age of mycelia

Figure (2): Percentage of degraded aflatoxin B1 by non-fragmented

mycelium of Aspergillus flavus and Aspergillus parasiticus.
 Azab, R.M. et al. 55

12

10

% of degraded AFB1 8

A. flavus
6
A. parasticus

0
5 6 7 8 9 10 11 12 13 14 15 16 17 18
Age of mycelia

Figure (3): Percentage of degraded aflatoxin B1 by fragmented

mycelium of Aspergillus flavus and Aspergillus parasiticus.

25

20
% of degraded AFB1

15
A. flavus
A. parasticus
10

0
5 6 7 8 9 10 11 12 13 14 15 16 17 18
Age of mycelia

Figure (4): Percentage of degraded aflatoxin B1 by non-heated cell free

system of Aspergillus flavus and Aspergillus parasiticus.
‫‪56‬‬ ‫‪Detection and Estimation of Aflatoxin B1 in Feeds and its‬‬
‫‪Biodegradation By Bacteria and Fungi‬‬

‫اﻟﻣﻠﺧص اﻟﻌرﺑﻰ‬
‫اﻟﻛﺷف ﻋن اﻻﻓﻼﺗوﻛﺳﯾن ب‪ ١‬وﺗﺣدﯾد ﻧﺳب ﺗواﺟده ﻓﻲ اﻟﻌﻠﯾﻘﺔ وﺗﺣﻠﻠﮫ اﻟﺑﯾوﻟوﺟﻰ‬
‫ﺑﺎﻟﺑﻛﺗﯾرﯾﺎ واﻟﻔطرﯾﺎت‬

‫راﻧﯾﺎ ﻣﻣﺗﺎز ﻋزب‪ ، ١‬واﺋل ﻣﺻطﻔﻰ ﺗوﻛل‪ ، ٢‬ﻋﺑداﻟرﺣﻣن ﻣﺻطﻔﻰ ﺣﻣﺎد‪ ،١‬ﻣﺣﻣد ﻛﻣﺎل اﺑو‬
‫‪٥‬‬
‫اﻟﻣﺟد‪ ، ٣‬ﺣﺳن ﻣﺻطﻔﻰ اﻷﺟرب‪ ، ٤‬ﻣﺣﻣد ﻛﻣﺎل رﻓﺎﻋﻰ‬
‫‪ ١‬ﻗﺳم اﻟﻔطرﯾﺎت ﺑﻣﻌﮭد ﺑﺣوث ﺻﺣﺔ اﻟﺣﯾوان‪ ٢ ،‬ﻗﺳم اﻟﻣﯾﻛروﺑﯾوﻟوﺟﯾﺎ ﺑﻛﻠﯾﺔ اﻟﺻﯾدﻟﺔ ﺟﺎﻣﻌﺔ اﻟﻘﺎھرة‬
‫‪ ٣،‬ﻗﺳم اﻟﻛﯾﻣﯾﺎء اﻟﺣﯾوﯾﺔ ﺑﻣﻌﮭد ﺑﺣوث ﺻﺣﺔ اﻟﺣﯾوان‪ ٤ ،‬ﻗﺳم اﻟﺻﺣﺔ واﻟرﻋﺎﯾﺔ اﻟﺑﯾطرﯾﺔ ﻛﻠﯾﺔ اﻟطب‬
‫اﻟﺑﯾطرى ﺟﺎﻣﻌﺔ اﻟﻘﺎھرة ‪ ٥ ،‬ﻗﺳم اﻟﻣﯾﻛروﺑﯾوﻟوﺟﯾﺎ ﻛﻠﯾﺔ اﻟطب اﻟﺑﯾطرى ﺟﺎﻣﻌﺔ اﻟﻘﺎھرة‬

‫‪ ٥٠٠‬أوﻋ ﻣﻛ ون ﻏ ذاﺋﻰ ﻟﻺﺻ ﺎﺑﺔ ﺑﻔط ر‬ ‫رى ﻟﻌ ﺔ ﻏدد ذاء‬

‫ﯾﻧ‬ ‫ﺗ م اﻟﻔﺣ ص اﻟﻔط‬
‫‪ ١٨٠‬ﻣ ن ﻓط ر‬ ‫زل رة‬ ‫ﺑرﺟﯾﻠس ﻓﻼﻓ س واﻻﺳ ﺑرﺟﯾﻠس ﺑ ﺎرازﺗﯾﻛس وﻗ د ﺗ م ﻋ ﻋﺗ‬ ‫اﻻﺳ‬
‫ر‬ ‫ن ﻓط‬
‫م ﻋ ﻣزل ‪٦٥‬‬ ‫ﺎﺗ‬ ‫ﺎر ﺑﯾﻧﻣ‬ ‫ت اﻻﺧﺗﺑ‬ ‫ﺎت ﺗﺣ‬ ‫ن اﻟﻌﯾﻧ‬ ‫سﻣ‬ ‫س ﻓﻼﻓ‬
‫ﺳم‬ ‫ﺎت ﻟ‬ ‫س اﻟﻌﯾﻧ‬ ‫اﻟﻌﯾﻧ ﻓﺣ ﺎت‪.‬ص ﻧﻔ‬‫ن ﻧﻔ د ﺗس م‬ ‫وﻗ‬ ‫ﺎرازﺗﯾﻛس ﻣ‬ ‫ﺑرﺟﯾﻠس ﺑ‬ ‫اﻻﺳ‬
‫‪٤٧.٤‬ن‪%‬ھ ذه اﻟﻌﯾﻧ ﺎت ﺗﺣﺗ وى ﻋﻠ ﻰ‬ ‫اﻷﻓﻼﺗوﻛﺳﯾن ب‪١‬وﻛﺎﻧت ﻧﺗﯾﺟ ﺔ اﻟﻔﺣ ص أن ﻣ‬
‫اﻷﻓﻼﺗوﻛﺳﯾن ب‪ ١‬وأن ‪ %١٩.٨‬ﻣن اﻟﻌﯾﻧﺎت اﻹﯾﺟﺎﺑﯾﺔ ﺗﺣﺗوى ﻋﻠﻰ أﻓﻼﺗوﻛﺳﯾن ب‪ ١‬ﻣﺎ‬
‫ﺑﯾن ‪ ٢٥ -١‬ﺟزء ﻓﻰ اﻟﺑﻠﯾون ﺑﯾﻧﻣﺎ ‪ %٩‬ﺗﺣﺗوى ﻋﻠﻰ أﻓﻼﺗوﻛﺳﯾن ب‪ ١‬ﻣﺎ ﺑﯾن ‪٥٠ -٢٦‬‬
‫‪١٠٠‬زء ﻓ ﻰ اﻟﺑﻠﯾ ون و ‪%٦.٤‬‬‫‪٨.٤‬وى ﻋﻠ ﻰ ﻣ ﺎ ﺑ ﯾن ‪ -٥١‬ﺟ‬ ‫ﺟزء ﻓﻰ اﻟﺑﻠﯾ ون وﺗﺣﺗ‬
‫وى‪%‬ﻋﻠ ﻰ ﻣ ﺎ ﺑ ﯾن ‪-٢٠١‬‬ ‫‪ ٢٠٠‬ﻓ ﻰ اﻟﺑﻠﯾ ونﺗﺣﺗو ‪٣.٨‬‬ ‫ﺟ‪ -‬زء‬ ‫ﻋﻠ ﻰ ﻣ ﺎ ﺑ ﯾن ‪١٠١‬‬
‫ون‪.‬اﺧﺗﺑ ﺎر اﻟﻌﺗ رات اﻟﻣﻌزوﻟ ﺔ ﻹﻓ راز اﻷﻓﻼﺗوﻛ ﺳﯾن ب‪١‬‬ ‫وﻋﻧ د‬‫‪٢٠٠٠‬ﺟزء ﻓﻰ اﻟﺑﻠﯾ‬
‫س وﻣ‪ ١٦‬ن‬ ‫ﺑرﺟﯾﻠس‬
‫ﻓﻼﻓ‬ ‫ﻻﺳ ا‬
‫ر‬ ‫ن ﻓط‬ ‫‪(%‬‬
‫‪٤٥)١٨٠‬رة ﻣ‬‫‪ ٨١‬ن ﻋﺗ‬ ‫د أن ﻣ‬ ‫وﺟ‬
‫ﺑرﺟﯾﻠس ﻟ دﯾﮭﺎ اﻟﻘ درة ﻋﻠ ﻰ إﻓ راز‬‫اﻻﺳ ﺎرازﺗﯾﻛس‬ ‫رة ﻣ ن ﻓط ﺑر‬ ‫‪(%٢٤,٦٢‬‬
‫‪ )٦٥‬ﻋﺗ‬
‫أﻓﻼﺗوﻛﺳﯾن ب‪.١‬‬
‫ﻛﻣﺎ ﺗم اﺧﺗﺑﺎر ﻣﻘدرة أرﺑﻌﺔ ﻋﺗرات ﻣن ﻣﯾﻛ روب اﻟﻼﻛﺗوﺑﺎﺳ ﻠس ﻋﻠ ﻰ إزاﻟ ﺔ ﺳ م‬
‫اﻷﻓﻼﺗوﻛﺳﯾن ب‪١‬‬
‫اﻟﻣﺳﺗﻧﺑت اﻟﺳﺎﺋل ﺑﻌ د ﻣﻌﺎﻟﺟﺗﮭ ﺎ ﺣرارﯾ ﺎ وﻛﯾﻣﯾﺎﺋﯾ ﺎ وﻗ د أظﮭ رت‬ ‫ﻣن‬
‫ﺣﻣ ﺿﯾﺎ أو ﺣرارﯾ ﺎ ﻟ دﯾﮭﺎ ﻣﻘ درة أﻛﺑ ر ﻋﻠ ﻰ إزاﻟ ﺔ ﺳ م‬ ‫أن اﻟﻌﺗ رات اﻟﻣﻌﺎﻟﺟاﻟﻧﺗ ﺔ ﺎﺋﺞ‬
‫ﺿﯾﺎ‪.‬‬ ‫ﺔ ﺣﻣ‬ ‫ﺎءة ﻣ ن اﻟﻣﻌﺎﻟﺟ‬ ‫ل ﻛﻔ‬ ‫ﺳﯾن ب‪١‬‬
‫ﺣرارﯾ ﺎ أﻗ‬ ‫ﺔ‬ ‫اﻷﻓﻼﺗوﻛ‬
‫اﻟﻣﻌﺎﻟﺟ‬ ‫ت‬ ‫وإن ﻛﺎﻧ‬
‫رت ﻧﺗ ﺎﺋﺞ اﺧﺗﺑ ﺎر ﻣﻘ درة اﻟﻧ ﺳﯾﺞ اﻟﻔط رى اﻟ ﺳﻠﯾم واﻟﻧ ﺳﯾﺞ اﻟﻔط رى اﻟﻣﺟ زء‬
‫واﻟرﺷﯾﺢ اﻟﻧﺎﺗﺞ ﻣن اﻟﻧﺳﯾﺞ اﻟﻔطرى اﻟﻣﺟزء ﻟﻛل ﻣن ﻓطر اﻻﺳﺑرﺟﯾﻠسﻓﻼﻓس و ﻓط ر‬
‫اﻷﻓﻼﺗوﻛ ﺳﯾن ب‪١‬أنوﺗﺟزﺋ ﺔ‬ ‫ﺎرازﺗﯾﻛس ﺎم ﻋﻠ ﻰ إزاﻟ ﺔ ﺳ م‬ ‫اﻟﻐﯾ ر ﺳ‬‫ﺑرﺟﯾﻠس‬
‫ﺑ‬ ‫اﻻﺳ‬
‫اﻟﻧﺳﯾﺞ اﻟﻔطرى ﯾزﯾد ﻣن إﻣﻛﺎﻧﯾﺔ اﻟﻌﺗرات اﻟﻣﺧﺗﺑ رة ﻋﻠ ﻲ إزاﻟ ﺔ ﺳ ماﻷﻓﻼﺗوﻛ ﺳﯾن ب‪١‬‬
‫وﻛﺎن اﻟرﺷﯾﺢ اﻟﺧﺎﻟﻲ ﻣن اﻟﺧﻼﯾﺎ ھو اﻷﻛﺛر ﻓﺎﻋﻠﯾﺔ ﻓﻲ إزاﻟﺔ ﺳﻣوم اﻷﻓﻼﺗوﻛ ﺳﯾن ب‪.١‬‬
‫م‬ ‫ﺔﺳ‬ ‫ﺑرﺟﯾﻠس ﻰ إزاﻟس‬
‫درة ﻓط اﻻﺳر ﻋﻠ ﻓﻼﻓ‬ ‫ﺔﺣأن ﻣﻘ‬ ‫أوﺿ‬
‫ﻛﻣ ت ﺎاﻟدراﺳ‬
‫اﻷﻓﻼﺗوﻛﺳﯾن ب‪ ١‬أﻛﺛر ﻣن ﻓطر اﻻﺳﺑرﺟﯾﻠس ﺑﺎرازﺗﯾﻛس‪.‬‬