Occurrence of toxigenic <i>Aspergillus flavus</i> in commercial Bulgur wheat

FARIA, Carla Bertechini; SANTOS, Fabiane Cristina dos; CASTRO, Fausto Fernandes de; SUTIL, Ariadne Ricieli; SERGIO, Luciana Marciano; SILVA, Milena Veronezi; MACHINSKI, Miguel; BARBOSA-TESSMANN, Ione Parra

doi:10.1590/1678-457X.09316

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Research Paper • Food Sci. Technol (Campinas) 37 (1) • Jan-Mar 2017 • https://doi.org/10.1590/1678-457X.09316 copy

Occurrence of toxigenic Aspergillus flavus in commercial Bulgur wheat

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Aflatoxins are mutagenic, carcinogenic, and teratogenic mycotoxins. The objective of this work was to study the presence of aflatoxigenic Aspergillus in commercial Bulgur wheat in the city of Maringá, Paraná, Brazil. Thirty samples of commercial Bulgur wheat, acquired in the period of August 2011 to January 2012, were evaluated. The enumeration analysis showed that samples had up to 273.3 CFU of molds and 133.3 CFU of aflatoxigenic Aspergillus per gram of wheat. Forty-two monosporic isolates were obtained and identified as Aspergillus flavus. The isolates were analyzed regarding their aflatoxigenic potential by culture in coconut milk agar; hydroxide vapor exposure; chromatography; and polymerase chain reaction (PCR) targeting genes that code enzymes of the aflatoxins synthesis pathway. Some of the isolates were confirmed to be aflatoxin producers and several of them presented a genetic profile of aflatoxin synthesis. The obtained results demonstrated that Bulgur wheat A. flavus contamination is concerning.

Keywords:
Aspergillus flavus; aflatoxins; Bulgur wheat; PCR

Month/ year	Sample	Molds CFU/g ^* * The data are given in CFU/g of Bulgur wheat sample and represent the average and the standard deviation obtained in the analysis of three dishes.	*Aspergillus* CFU/g ^* * The data are given in CFU/g of Bulgur wheat sample and represent the average and the standard deviation obtained in the analysis of three dishes.	Isolates of A. flavus	GenBank Acession number ^# # The isolates were identified by using the ITS region.
Aug/2011	1	180 ± 20	40.0 ± 13.3	1-1	KT329248
				1-2	KT329249
				1-3	KT329250
	2	273.3 ± 71.1	73.3 ± 44.4	2-1	KT329251
				2-2	KT329252
				2-3	KT329253
	3	213.3 ± 17.8	80.0 ± 33.3	3-1	KT329254
				3-2	KT329255
				3-3	KT329256
	4	0 ± 0	0 ± 0	-
	5	0 ± 0	0 ± 0	-
Sep/2011	6	33.3 ± 22.2	33.3 ± 22.2	6-2	KT329257
	6	33.3 ± 22.2	33.3 ± 22.2	6-3	KT329258
	7	223.3 ± 35.6	123.3 ± 4.4	7-1	KT329259
				7-2	KT329260
				7-3	KT329261
	8	73.3 ± 31,1	0 ± 0	-
	9	26.7 ± 4.4	0 ± 0	-
	10	16.7 ± 4.4	3.3 ± 4.4	10-3	KT329262
Oct/2011	11	43.3 ± 8.9	16.7 ± 4.4	11-1	KT329263
	11	43.3 ± 8.9	16.7 ± 4.4	11-2	KT329264
	12	53.3 ± 11.1	13.3 ± 4.4	12-1	KT329265
				12-2	KT329266
				12-3	KT329267
	13	26.77 ± 22.2	0 ± 0	-
	14	16.7 ± 8.9	3.3 ± 4.4	14-2	KT329268
	15	226.7 ± 24.4	133.3 ± 4.4	15-1	KT329269
	15	226.7 ± 24.4	133.3 ± 4.4	15-2	KT329270
Nov/2011	16	136.7 ± 37.8	16.7 ± 11.1	16-1	KT329271
	16	136.7 ± 37.8	16.7 ± 11.1	16-3	KT329272
	17	16.7 ± 4.4	10.0 ± 6.7	17-2	KT329273
	17	16.7 ± 4.4	10.0 ± 6.7	17-3	KT329274
	18	110.0 ± 6.7	83.3 ± 15.6	18-1	KT329275
	18	110.0 ± 6.7	83.3 ± 15.6	18-2	KT329276
	19	40.0 ± 6.7	13.3 ± 4.4	19-1	KT329277
				19-2	KT329278
				19-3	KT329279
	20	13.3 ± 8.9	3.3 ± 4.4	20-1	KT329280
Dec/2011	21	0 ± 0	0 ± 0	-
	22	66.7 ± 4.4	46.7 ± 11.1	22-1	KT329281
	22	66.7 ± 4.4	46.7 ± 11.1	22-3	KT329282
	23	10.0 ± 13.3	0 ± 0	-
	24	90.0 ± 20.0	20.0 ± 6.7	24-1	KT329283
	24	90.0 ± 20.0	20.0 ± 6.7	24-2	KT329284
	25	46.7 ± 4.4	0 ± 0	-
Jan/2012	26	210.0 ± 40.0	43.3 ± 15.6	26-1	KT329285
	26	210.0 ± 40.0	43.3 ± 15.6	26-3	KT329286
	27	36.7 ± 11.1	0 ± 0	-
	28	90.0 ± 120.0	0 ± 0	-
	29	273.3 ± 168.9	0 ± 0	-
	30	133.3 ± 24.4	106.7 ±11.1	30-1	KT329287
				30-2	KT329288
				30-3	KT329289
Total		87.8 ± 22.7^a	28.8 ± 7.32^b

Isolate	Fluorescence in CMA^* * Analysis in triplicate.	Ammonium hydroxide vapour^* * Analysis in triplicate.	TLC		HPLC	PCR
Isolate	Fluorescence in CMA^* * Analysis in triplicate.	Ammonium hydroxide vapour^* * Analysis in triplicate.	MGA	CMA	YES	apa-2	omt-A	ver-1	nor-1
1-1	-	-	-	-	-	+	+	+	+
1-2	-	-	-	-	-	+	+	+	+
1-3	-	-	-	-	-	-	-	+	-
2-1	-	-	-	-	-	+	+	+	+
2-2	-	-	-	-	-	+	+	+	+
2-3	-	-	-	-	-	+	+	+	+
3-1	-	-	-	-	-	-	+	+	+
3-2	-	-	-	-	-	+	+	+	+
3-3	-	-	-	-	-	+	+	+	+
6-2	-	-	-	-	-	-	+	+	+
6-3	-	-	-	-	-	-	+	+	+
7-1	-	-	-	-	-	+	+	+	+
7-2	-	-	-	-	-	+	+	+	+
7-3	-	-	-	-	-	+	+	+	+
10-3	-	-	-	-	-	+	+	+	+
11-1	-	-	-	-	-	+	+	+	+
11-2	-	-	-	-	-	-	+	+	+
12-1	-	-	-	-	-	+	+	+	+
12-2	-	-	-	-	-	+	+	+	+
12-3	-	-	-	-	+	+	+	+	+
14-2	-	-	-	-	-	+	+	+	+
15-1	-	-	-	-	+	+	+	+	+
15-2	-	-	-	-	-	-	+	+	+
16-1	-	-	-	-	-	+	+	+	+
16-3	-	-	-	-	-	+	+	+	+
17-2	-	-	-	-	-	-	+	+	+
17-3	-	-	-	-	-	+	+	+	+
18-1	-	-	-	-	-	+	+	+	+
18-2	-	-	-	-	-	+	+	+	+
19-1	-	-	-	-	-	+	+	+	+
19-2	-	-	-	-	-	+	+	+	+
19-3	-	-	-	-	-	+	+	+	+
20-1	-	-	-	-	+	+	+	+	+
22-1	-	-	-	-	-	-	+	+	-
22-3	-	-	-	-	-	+	+	+	+
24-1	+	+	-	+	-	+	+	+	+
24-2	-	-	-	-	-	-	-	+	-
26-1	-	-	-	-	-	+	+	+	+
26-3	-	-	-	-	-	+	+	+	+
30-1	-	-	-	-	+	-	-	+	+
30-2	-	-	-	-	-	+	+	+	+
30-3	-	-	-	-	-	-	+	+	-
443^# # A. parasiticus UEM 443 and A. flavus NRRL 5940.	+	+	+	+	+	+	+	+	+
5940^# # A. parasiticus UEM 443 and A. flavus NRRL 5940.	+	+	+	+	+	+	+	+	+

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[1] *Corresponding author: ipbtessmann@uem.br