Determination of total a atoxin in rice consumption in

Yasuj, Iran

Ali Mousavizadeh

, Amir Peikar

, Ahmad Hoseinian

, Emambakhsh Ghaydie

and

Azizollah Pourmahmoudi

PhD in Epidemiology, Social Determinants of Health Research Center, Yasuj University of Medical Sciences

Yasuj, Iran

Laboratory Manager Of Food And Drug Institute Attached To Yasuj University Of Medical Science, Iran

Expert Laboratory Of Food And Drug Institute Attached To Yasuj University Of Medical Science, Iran

Experimental Expert of Food And Drug Institute Attached To Yasuj University Of Medical Science, Iran

Ph.D – Nutrition, Determinants of Nutrition,School of Health, Yasuj University of Medical Sciences, Yasuj, Iran

ABSTRACT

A atoxins are the secondary metabolitesof Aspergillus  avus and Aspergillus parasiticus which are often produced

by molds. The consumption of food contaminated with a atoxin poison, which is associated with many humans and

A atoxicosis, in some cases can lead to death.Nowadays, rice is the foodstuff for half of theworld’spopulation. Rice is

exposed to fungal and a atoxin contaminations like other cereal crops. This study aimed toinvestigate and compare

the amount of a atoxin and the level of toxin in rice samples in Yasuj. In this study, 45rice samples collected from

Yasuj supermarketsand the amount of a atoxin contamination were tested by Elisa. The results showed that total

a atoxincontamination in all samples was lower than the national standard in Iran. The average amount of total

a atoxin in types such as Gerdeh, Champa, Shamim and Fajr, is6.53- 6.14- 4.54and 6.12 ng /g, respectively. Compar-

ing the results of this study and other studies, it can be concluded that rice examined in this study is more desirable.

KEY WORDS: TOTAL AFLATOXIN, RICE, ELISA

195

ARTICLE INFORMATION:

*Corresponding Author: pourmahmoudi@gmil.com

Received 27

Dec, 2016

Accepted after revision 2

March, 2017

BBRC Print ISSN: 0974-6455

Online ISSN: 2321-4007

Thomson Reuters ISI ESC and Crossref Indexed Journal

NAAS Journal Score 2017: 4.31 Cosmos IF : 4.006

reserved.

Online Contents Available at: http//www.bbrc.in/

Biosci. Biotech. Res. Comm. Special Issue No 1:195-198 (2017)

196 DETERMINATION OF TOTAL AFLATOXIN IN RICE CONSUMPTION IN YASUJ, IRAN BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

Ali Mousavizadeh et al.

INTRODUCTION

Rice is one of the most important crops. Grain dark is

the staple food for half of the world’s population (Payan,

2011). Rice cultivation has existed in Iran since the

Achaemenid (Khodabande, 2010). It is cultivated widely

in certain areas of the world such as China, India, Japan,

Uruguay and parts of South Africa (Payan, 2011). Many

agricultural products from the cultivation to consumption

are prone to contaminate with fungi. If the fungipollu-

tion-causing strains of toxin,it may produce mycotoxin

in some stage of its growth as a secondary metabolite.

(Reza, Tahereh, Alireza, & bahar, 2011) Liu et al, 2015).

Despite numerous studies during the  rst half of the

last century, only in the 1960s, itwas found that metabo-

lites of fungi are responsible for the illness and death

of livestock. Currently, it is well established that toxic

mold metabolites are responsible for many epidemics in

human and animal population (Reza, Tahereh, Alireza,

& bahar, 2011). Mycotoxins are a natural group of toxic

compounds produced by several species of fungi. These

compounds have different chemical structuresandlow

molecular weight that are found in a large number of

agricultural products and food. In general, many types

of grains, oilseeds, nuts and dried fruit are prone to fun-

gal pollution(Majid, et al., 2008). The most important

mycotoxins involve a atoxins, patulin, ochratoxin A

and zearalenone (Abdel-Wahhab & Kholif, 2010).

A atoxins are mycotoxins produced by the Aspergil-

lus species and some species of Penicillium and Asper-

gillus  avus (FinkGrernmels, 1999). Different types of

a atoxin are: AFP1, AFG2, AFG1, AFM2, AFM1, AFB2,

AFB1 and AFQ1 that a atoxin B1, biologically is the

most active type between theknown a atoxins (Kazemi,

Mohtadinia, Mahdavi, Akbari, & Salehpour, 2008). The

most important factors in a atoxin production include

temperature, humidity, oxygen concentration, substrate,

PH nutrients, microbial interactions, the presence or

absence of inhibitors such as organic acids as well as

mechanical damage (Amanlou, Rezaei Khkha, Ramazani,

& Meyer, 2014, Liu et al, 2015).

Kohgiluyeh and Boyer-Ahmad is one of the prov-

inces, which has water resources and favorable cli-

matic conditions for rice cultivation (Nader, Kobra, &

Mohammad Javad, 2008). Six common varieties of rice

in this province are Champa, Gerdeh, Tropical Champa,

Shamim, Lenjan and Tarom (Cassel, Campbell, Draper, &

Epperson, A atoxins: Hazards in grain/a atoxicosis and

livestock, 2001), (Arafa, Bloomer, Wilson, Simpson, &

Harms, 1981). Since the review of total a atoxin has not

been conducted in this province and according to the

importance of total a atoxin monitoring in rice, thisre-

search evaluates the total a atoxin in the rice produced

in Kohgiluyeh and Boyer-Ahmad.

MATERIALS AND METHODS

In this study, the rice sample was randomly collected

from Yasuj shops, accordingto the sampling method of

crops.

TOTAL AFLATOXIN ANALYSIS BY ELISA

In recent years ELISA method is used for the determi-

nation of mycotoxins. The ELISA method as a gen-

eral technique for the early diagnosis and detection of

mycotoxins with advantages such as sensitivity, sample

preparation and easy operation, safety, high accuracy

and time saving can be noted. This chemical method

used antigen-antibody technique. This method is sensi-

tive, fast and relatively inexpensive(Institute of Stand-

ards and Industrial Research of Iran, 2001)-(Khosravi,

1998).Total A atoxin competitive immunoassay kit

EuroProxima B.V of the Netherlands was used to deter-

mine how much of the toxin existed in their food.

Extraction and test methods for mycotoxins in the sam-

ples were performed according to the manufacturer’s

instructions.

SAMPLE PREPARATION

10 g ofthe rice powder sample was homogenized with

50ml of acetonitril in room temperature for 30 min-

utes was completely shaken. The extract was  ltered by

No. 42 Whatman paper and then 50 ml of the  ltrate

was diluted with 150 ml of buffer solution.100 ml of

diluted extract was used per sink in the test.The rest was

homogenized for 10 min at roomtemperature and then

the resultant deposit was centrifuged.An aliqout (100

µl) of the supernatant was diluted with600 µl of phos-

phate buffer at pH = 7.2. An aliquot of thissolution (50

µl) or standard solution (50 µl), 50 µl of thea atoxin-

peroxidase conjugate and 50 µl of the mouseantibody

solution against alfatoxin were added to eachsink of the

used plate. The determination was replicatedthree times.

The samples were incubated for 30 min at room tem-

perature in the darkness. The free and peroxidase-com-

bined a atoxins compete for the combining site with

antibodies to mouse antibodies immobilized on theplate.

Next, the plate was emptied and washed  ve times with

phosphate buffer at pH = 7.2. Then, 50 µl of tetramethyl

benzidineAnd 50 µl of urea peroxide were added and

incubated again for 30 min in darkness. The reactionwas

terminated by adding 100 µl of the stop reagent. Theab-

sorbance of the solution was measured at a wavelength

of450 nm, using an ELISA reading apparatus. The con-

tent of a atoxins was calculated using the previously

preparedstandard curve.

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS DETERMINATION OF TOTAL AFLATOXIN IN RICE CONSUMPTION IN YASUJ, IRAN 197

Ali Mousavizadeh et al.

RESULTS AND DISCUSSION

The results are shown in Table 1. The  ndings of this

study show that of the 45 samples all samples had total

a atoxin contamination, but it was lower than the stand-

ard limit. The limit of the toxin in different countries var-

ies from 30 g / kg to50 g food. For example, in Iran it

is 30 g / kg of food material (Food corporation of India,

2011)-(Jonathan, et al., 2004).The maximum amount of

a atoxin 6.9 and the lowest amount of poison was 0.06

.The average amount of poison in the tested samples was

measured 5.83 g / kg. Average total a atoxin concen-

tration of toxin in the samples of rice Gerdeh, Champa,

Shamim and Fajr, 6.53,6.14,4.54 and 6.12 g / kg,

respectively.The maximum data of The Gerdeh, Champa,

Shamim and Fajr, sample, 6.62,6.42,6.68 and 6.9 g / kg

respectivelyand minimum amount of them 6.42,5.24,0.06

and 5.09 respectively (Table 1). Figure 1 shows Average

total a atoxin in the rice samples.

Rice is among the food items in the basket of Irani-

ans. Due to the high consumption of rice by the Iranians

considering quality and health, it can have an important

role in maintaining and promoting overall health.How-

ever, due to the high volume of product, product diver-

sity, different cultures, different weather conditions and

other causes of contamination and harmful substances,

a variety of factors exist. Among the most important

and common infections, fungal infections and contam-

inated mold can be noted. A atoxins and Ochratoxin

by a group of the secondary metabolites of fungi that

are particularly Aspergillus arise Azqarch. Studies have

shown that post-harvest processing as well as applying

proper drying and storage to reduce pollution and pre-

vent its spread areis is required (Rustom, 1997)-(Sama-

rajeewa, Sen, & Cohen, 1990). In a study by Kazemi

et al on rice East Azerbaijan, 23% of the samples were

infected. (Department of Planning & Economic Infor-

mation and Communication Technology Center, 2013-

2014). In another study by Rezai on imported rice in

Zabol, 6.27% of thesamples hadfungal infection (Center

for Statistics and Information Technology Ministry of

Agriculture , 2013-2014).

A recent study by (Liu et al, 2015) on the 370 rice

schema from six provinces in China was conducted to

determine a atoxin and Ochratoxin A 5/63 percent and

9.4 percent, The results showed that the samples con-

tained Ochratoxin a 4.1% of samples were a atoxin

and a atoxin and 3.0% of samples above the limit

Ochratoxin a Europe Union (Farsiani, Marziye, Mahsa,

Mohamad reza, & Mahmoud, 2014).Figure 1 shows the

present study.Most of that total a atoxin investigate the

prevalence of less than 30 ng per mg, based on Table 1,

Table 1. Analysis of total a atoxin in the samples of

rice in the city of Yasooj.

Mim Max SD Average number of

samples

Samples

6.42 6.63 0.09 6.53 7 Gerdeh

5.24 6.42 0.21 6.14 17 Champa

0.06 6.48 2.02 4.54 11 Shamim

5.09 6.9 0.58 6.12 9 Fajr

FIGURE 1. Comparison of the average total a atoxin in the samples with standard limit

Ali Mousavizadeh et al.

198 DETERMINATION OF TOTAL AFLATOXIN IN RICE CONSUMPTION IN YASUJ, IRAN BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

the average total a atoxin in samples examined 83/5

on the ng/mg. In a study by Elena Suárez-Bonnet et al

(2013), based on the total a atoxin contamination (B1,

G1, B2, and G2) in rice from Mexico and Aspanya, the

results showed that the average total a atoxin of Span-

ish rice was 37 / 3 micrograms per kg, in the range of 6.1

to 1383 mg/ kg (SuárezBonnet, et al., 2013).

CONCLUSION

Comparing the results of this study and studies in the

country and other countries it can be concluded that the

rice examined in this study is more desirable. However,

the potential pollution. to the fungus and toxins result-

ing from it and universal standards in the harvest of

all itemsshould beapplied to the provide perfect condi-

tions for transportation and time- consuming mainte-

nance. Another important point is that the cultivation

and harvest time or until you quit at the end of bran or

less is consumed, the possibility of less contamination

is more. The role of temperature in mold growth and

toxin production after harvesting isproven and it must

be maintained at the right temperature as much as the

mold growth is prevented.

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