Effect of Temperature, Water Activity and Incubation Time on Trichothecene Production by Fusarium cerealis Isolated from Durum Wheat Grains.

Fusarium cerealis is a causal agent of Fusarium Head Blight in wheat, and it produces both deoxynivalenol (DON) and nivalenol (NIV). Nevertheless, the effect of environmental factors on the growth and mycotoxin production of this species has not been studied so far. The objective of this study was to investigate the impact of environmental factors on the growth and mycotoxin production of F. cerealis strains. All strains were able to grow in a wide range of water activity (aW) and temperatures, but their mycotoxin production was influenced by strain and environmental factors. NIV was produced at high aW and temperatures, while optimal conditions for DON production were observed at low aW. Interestingly, some strains were able to simultaneously produce both toxins, which could pose a more significant risk for grain contamination.

Production of type-B trichothecenes by Fusarium meridionale, F. graminearum, and F. austroamericanum in wheat plants and rice medium.

Food security goes beyond food being available; the food needs to be free of contaminants. Trichothecenes mycotoxins, produced by Fusarium fungus, are. among the most frequently found contaminants of wheat. In this study, we evaluated the production of trichothecenes Deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON), and nivalenol (NIV) by Fusarium meridionale, F. austroamericanum, and F. graminearum grown in wheat plants and rice medium. Fusarim meridionale was efficient only in the production of NIV (production range (pr) from 1340 to 2864 µg kg-1 in wheat plant), and F. austroamericanum in the production of 3-AcDON (pr from 50 to 192 µg kg-1 in wheat plant, and from 986 to 7045 µg kg-1 in rice medium) and DON (pr from 4076 to 13,701 µg kg-1 in wheat plant, and from 184 to 43,395 µg kg-1 in rice medium). Already, F. graminearum was efficient in the production of 3-AcDON only in rice medium (pr from 81 to 2342 µg kg-1), 15-AcDON in wheat plant (pr from 80 to 295 µg kg-1) and in rice medium (pr from 436 to 8597 µg kg-1), and DON also in wheat plant (pr from 7746 to 12,046 µg kg-1) and in rice medium (pr from 695 to 49,624 µg kg-1). The specificity of F. meridionale in the production of NIV but not the production of DON could generate a food security problem in regions where this species occurs and the amounts of NIV in grains and derivatives are not regulated in the food chain, as in Brazil.

Survey of Freshly Harvested Oat Grains from Southern Brazil Reveals High Incidence of Type B Trichothecenes and Associated Fusarium Species.

The current study investigated the fungal diversity in freshly harvested oat samples from the two largest production regions in Brazil, Paraná (PR) and Rio Grande do Sul (RS), focusing primarily on the Fusarium genus and the presence of type B trichothecenes. The majority of the isolates belonged to the Fusarium sambucinum species complex, and were identified as F. graminearum sensu stricto (s.s.), F. meridionale, and F. poae. In the RS region, F. poae was the most frequent fungus, while F. graminearum s.s. was the most frequent in the PR region. The F. graminearum s.s. isolates were 15-ADON genotype, while F. meridionale and F. poae were NIV genotype. Mycotoxin analysis revealed that 92% and 100% of the samples from PR and RS were contaminated with type B trichothecenes, respectively. Oat grains from PR were predominantly contaminated with DON, whereas NIV was predominant in oats from RS. Twenty-four percent of the samples were contaminated with DON at levels higher than Brazilian regulations. Co-contamination of DON, its derivatives, and NIV was observed in 84% and 57.7% of the samples from PR and RS, respectively. The results provide new information on Fusarium contamination in Brazilian oats, highlighting the importance of further studies on mycotoxins.

Fusarium cerealis causing Fusarium head blight of durum wheat and its associated mycotoxins.

Fusarium Head Blight (FHB) is a very important fungal disease that affects small grain cereals worldwide. This disease not only causes yield loses but also crops contamination with mycotoxins such as deoxynivalenol (DON) and nivalenol (NIV). Species within the Fusarium graminearum species complex have been described as the main causal agents of this disease, however lately there have been few reports of Fusarium cerealis causing the disease in wheat and barley in different parts of the world. This study evaluated the aggressiveness of F. cerealis to durum wheat cultivars and also mycotoxin production in planta. Moreover, the mycotoxin profile of F. cerealis strains was characterized molecularly and chemically. All durum wheat cultivars showed typical FHB symptoms but the disease severity varied among them in levels up to 66%. In addition, seventeen different compounds were detected in the infected heads including DON, NIV and nivalenol-3-ß-d-glucose (NIV3G). NIV was detected in all cultivars and was the most produced mycotoxin with levels ranging from 1.04 to 6.8 mg/kg. On the other hand, the molecular analysis of F. cerealis strains showed that all of them possessed NIV genotype while the chemical assessment showed that the strains were able to produce not only this toxin in vitro but also DON, zearalenone and other twenty-one secondary metabolites. The increasing incidence of F. cerealis and the possible contamination of crops with the mycotoxins that it produces are of great concern for food security and world cereal trade since it has been reported that NIV is more toxic for humans and animals than DON.

The Dominance of Fusarium meridionale Over F. graminearum Causing Gibberella Ear Rot in Brazil May Be Due to Increased Aggressiveness and Competitiveness.

In Brazil, Gibberella ear rot (GER) of maize is caused mainly by Fusarium meridionale, whereas F. graminearum is a minor contributor. To test the hypothesis that F. meridionale is more aggressive than F. graminearum on maize, six experiments were conducted in the south (summer) and one in the central-south (winter), totaling seven conditions (year × location × hybrid). Treatments consisted of F. graminearum or F. meridionale (two isolates of each) inoculated once 4 days after silk, inoculated sequentially and alternately (F. graminearum → F. meridionale or F. meridionale → F. graminearum) 6 days apart, or (in the central-south) inoculated sequentially without alternating species (F. meridionale → F. meridionale or F. graminearum → F. graminearum). Overall, severity was two times greater in the south (37.0%), where summer temperatures were warmer (20 to 25°C) than in central-south. In the south, severity was greatest in F. meridionale treatments (67.8%); followed by F. meridionale → F. graminearum (41.1%), then F. graminearum → F. meridionale (19.4%), and lowest in F. graminearum (2.1%), suggesting an antagonistic relationship. In the central-south (15 to 20°C), severity was generally higher in the sequential nonalternating inoculation treatments (F. meridionale → F. meridionale or F. graminearum → F. graminearum) than when either species was inoculated only once. Only nivalenol (NIV) or deoxynivalenol was detected when F. meridionale or F. graminearum, respectively, was inoculated singly, or sequentially with no alternation. Both toxins were found in grains harvested from the F. meridionale → F. graminearum treatment, whereas only NIV was found in kernels from the F. graminearum → F. meridionale treatment, suggesting that F. meridionale was more competitive than F. graminearum in coinoculations. The dominance of F. meridionale as a cause of GER in Brazil may be due in part to its higher aggressiveness and competitiveness compared with F. graminearum.

Effect of water activity and temperature on growth and trichothecene production by Fusarium meridionale.

Fusarium meridionale has been frequently isolated from soybean in Argentina and showed similar pathogenicity as F. graminearum sensu stricto. However, no data on their growth and mycotoxin production under different environmental conditions are yet available. The aims of this study were: to determine the effect of temperature, water activity (aW) and strain on growth of F. meridionale and to evaluate deoxynivalenol (DON) and nivalenol (NIV) production in a soybean based medium. The results showed that optimal conditions for F. meridionale growth were at 25 °C and 0.98-0.99 aW. Deoxynivalenol production was favored at 25 °C and 0.96 aW while NIV production was strain-dependent, being 30 °C and 0.98 aW optimal conditions for F. meridionale B2300 strain and 20 °C and 0.98 aW for F. meridionale F5043 and F. meridionale 5048 strains. These conditions are similar to those observed at pre-harvest stage in soybean crop, thus control strategies need to be considered to reduce the risk of the occurrence of DON and NIV in harvested grains.

Deoxynivalenol alone or in combination with nivalenol and zearalenone induce systemic histological changes in pigs.

Deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEA) are mycotoxins commonly produced by Fusarium species. The purpose of the present study was to investigate the effects of DON alone and in combination with NIV and ZEA on several parameters including weight gain and histological aspects of pigs submitted to chronic intoxication. Twenty, 5-week-old piglets received for 28 days one of the following diets: a control diet, a diet mono- contaminated with DON (1.5mg/kg), a diet multi-contaminated with DON (2mg/kg)+NIV (1.3mg/kg)+ZEA (1.5mg/kg) or a diet contaminated with DON (3mg/kg)+NIV (1.3mg/kg)+ZEA (1.5mg/kg). Animals fed the multi-contaminated diets presented a significant decrease in weight gain over the total period. The chronic ingestion of the contaminated diets induced a significant increase on histological changes on the intestine, liver and lymphoid organs. In addition, a significant increase on lymphocyte apoptosis was observed in lymph nodes and spleen in the animals receiving the contaminated diets. These data provide a better understanding of the possible effects of Fusarium toxins, alone or in combinations on the morphology of the intestine and lymphoid organs, which would contribute to the risk assessment of these toxins.

Incidence and multiplex PCR based detection of trichothecene chemotypes of Fusarium culmorum isolates collected from freshly harvested Maize kernels in Southern India.

Hundred Fusarium culmorum strains, isolated from freshly harvested maize grain samples from Southern parts of India, were incubated in czapek-dox medium and analyzed for trichothecene (DON/NIV) production. The mPCR assay was standardized targeting trichothecene metabolic pathway genes viz., Tri6, Tri7, Tri13 for detection of trichothecene (DON/NIV) chemotypes and rDNA gene for specific detection of F. culmorum species. Primers for targeted genes were designed and used to predict whether these isolates could produce deoxynivalenol/nivalenol, 94 isolates were able to produce DON/NIV by mPCR assay. Chemical analysis of DON/NIV was carried out for mPCR positive isolates by high performance-thin layer chromatography (HPTLC). To check the practical usefulness of developed mPCR assay, 150 field samples of maize were evaluated and results were compared with conventional HPTLC method. Out of 150 samples, 34% samples stayed as a positive for NIV contamination whereas 44% were found to have deoxynivalenol contamination. Moreover, mPCR results are equivocally matched with the HPTLC chemical analysis for field samples. Chemotyping of F. culmorum isolates were reported for the first time from India, and highlights the important potential of F. culmorum to contaminate maize with DON/NIV.

Regional differences in species composition and toxigenic potential among Fusarium head blight isolates from Uruguay indicate a risk of nivalenol contamination in new wheat production areas.

Members of the Fusarium graminearum species complex (FGSC) are the primary cause of Fusarium head blight (FHB) of wheat, and frequently contaminate grain with trichothecene mycotoxins that pose a serious threat to food safety and animal health. The species identity and trichothecene toxin potential of 151 FGSC isolates collected from wheat in Uruguay were determined via multilocus genotyping. Although F. graminearum with the 15ADON trichothecene type accounted for 86% of the isolates examined, five different FGSC species and all three trichothecene types were identified in this collection. This is the first report of Fusarium asiaticum, Fusarium brasilicum, Fusarium cortaderiae, and Fusarium austroamericanum from Uruguay. In addition, we observed significant (P<0.001) regional differences in the composition of FGSC species and trichothecene types within Uruguay. Isolates of F. graminearum with the 15ADON type were the most prevalent in western provinces (95%), while F. asiaticum (43%) and the NIV type (61%) predominated in the new wheat production zone in Cerro Largo along Uruguay's eastern border with Brazil. F. graminearum isolates (15ADON type) were significantly (P<0.005) more aggressive on wheat than were isolates from the other species examined (NIV or 3ADON types). However, F. graminearum isolates (15ADON type) were significantly (P<0.05) more sensitive to tebuconazole than isolates from other species (NIV type). These results document substantial heterogeneity among the pathogens responsible for FHB in Uruguay. In addition, the regional predominance of the NIV trichothecene type is of significant concern to food safety and indicates that additional monitoring of nivalenol levels in grain may be required.

Incidence and multiplex PCR based detection of trichothecene chemotypes of Fusarium culmorum isolates collected from freshly harvested Maize kernels in Southern India

Hundred Fusarium culmorum strains, isolated from freshly harvested maize grain samples from Southern parts of India, were incubated in czapek-dox medium and analyzed for trichothecene (DON/NIV) production. The mPCR assay was standardized targeting trichothecene metabolic pathway genes viz., Tri6, Tri7, Tri13 for detection of trichothecene (DON/NIV) chemotypes and rDNA gene for specific detection of F. culmorum species. Primers for targeted genes were designed and used to predict whether these isolates could produce deoxynivalenol/nivalenol, 94 isolates were able to produce DON/NIV by mPCR assay. Chemical analysis of DON/NIV was carried out for mPCR positive isolates by high performance-thin layer chromatography (HPTLC). To check the practical usefulness of developed mPCR assay, 150 field samples of maize were evaluated and results were compared with conventional HPTLC method. Out of 150 samples, 34% samples stayed as a positive for NIV contamination whereas 44% were found to have deoxynivalenol contamination. Moreover, mPCR results are equivocally matched with the HPTLC chemical analysis for field samples. Chemotyping of F. culmorum isolates were reported for the first time from India, and highlights the important potential of F. culmorum to contaminate maize with DON/NIV.

Incidence and multiplex PCR based detection of trichothecene chemotypes of Fusarium culmorum isolates collected from freshly harvested Maize kernels in Southern India

Hundred Fusarium culmorum strains, isolated from freshly harvested maize grain samples from Southern parts of India, were incubated in czapek-dox medium and analyzed for trichothecene (DON/NIV) production. The mPCR assay was standardized targeting trichothecene metabolic pathway genes viz., Tri6, Tri7, Tri13 for detection of trichothecene (DON/NIV) chemotypes and rDNA gene for specific detection of F. culmorum species. Primers for targeted genes were designed and used to predict whether these isolates could produce deoxynivalenol/nivalenol, 94 isolates were able to produce DON/NIV by mPCR assay. Chemical analysis of DON/NIV was carried out for mPCR positive isolates by high performance-thin layer chromatography (HPTLC). To check the practical usefulness of developed mPCR assay, 150 field samples of maize were evaluated and results were compared with conventional HPTLC method. Out of 150 samples, 34% samples stayed as a positive for NIV contamination whereas 44% were found to have deoxynivalenol contamination. Moreover, mPCR results are equivocally matched with the HPTLC chemical analysis for field samples. Chemotyping of F. culmorum isolates were reported for the first time from India, and highlights the important potential of F. culmorum to contaminate maize with DON/NIV.(AU)

Natural occurrence of nivalenol and mycotoxigenic potential of Fusarium graminearum strains in wheat affected by head blight in Argentina

The principal agents of Fusarium head blight in the main cropping area of Argentina were investigated in heavily infected samples. The ability of the isolates to produce trichothecenes was determined by GC and HPLC. Fusarium graminearum was the predominant species and of 33 isolates, 10 produced deoxinivalenol (DON) (0.1- 29 mg kg-1), 13 produced both deoxinivalenol (1.0- 708 mg kg-1) and nivalenol (0.1- 6.2mg kg-1), 12 produced 3-acetyldeoxinivalenol (0.1- 14 mg kg-1), 13 produced 15-acetyldeoxinivalenol (0.1- 1.9 mg kg-1), 10 produced Fusarenone X (0.1- 2.4 mg kg-1) and 7 produced zearalenone (0.1- 0.6 mg kg-1). These results suggest that F. graminearum strains isolated from the wheat growing regions in Argentina belong to DON chemotype. Although some strains produced both deoxinivalenol and nivalenol, nivalenol was produced in lower levels. The natural occurrence of nivalenol in wheat affected by head-blight collected in the main production area during two years (2001-2002) was also determined. From 19 samples 13 were contaminated with deoxinivalenol in a range of 0.3 to 70 mg kg-1and 2 samples with both deoxinivalenol (7.5 and 6.7 mg kg-1) and nivalenol (0.05 and 0.1 mg kg-1), respectively. This is the first report of natural occurrence of nivalenol in wheat cultivate in Argentina.

O principal causador de giberela no trigo na Argentina e sua capacidade de produzir tricotecenos foram estudados por GC e HPLC em amostras altamente infectadas. A espécie predominante foi Fusarium graminearum, sendo que de um total de 33 isolados, 10 produziram deoxinivalenol (0,1-29 mg kg -1), 13 produziram deoxinivalenol (1,0-708 mg kg-1) e nivalenol (0,1-6,2 mg kg-1), 12 produziram 3-acetildeoxinivalenol (0,1-14 mg kg-1), 13 produziram 15-acetildeoxinivalenol (0,1-1,9 mg kg-1), 10 produziram fusarenona X (0,1- 2,4 mg kg-1) e 7 produziram zearalenona (0,1- 0,6 mg kg-1). Esses resultados sugerem que as cepas de F. graminearum isoladas de trigo cultivado na Argentina pertencem ao quimiotipo DON. Embora algumas cepas tenham produzido tanto DON quanto NIV, NIV foi produzido em quantidade inferior ao DON. A ocorrência natural de nivalenol em trigo afetado pela giberela coletado na principal área de produção durante dois anos (2001-2002) foi também determinada. De 19 amostras, 13 estavam contaminadas com deoxinivalenol na faixa de 0,3 a 70 mg kg-1 e 2amostras continham tanto deoxinivalenol (7,5 e 6,7 mg kg-1) quanto nivalenol (0,05 e 0,1 mg kg-1), respectivamente. Esse é o primeiro relato da ocorrência de nivalenol em trigo cultivado na Argentina.

Natural occurrence of nivalenol and mycotoxigenic potential of Fusarium graminearum strains in wheat affected by head blight in Argentina.

The principal agents of Fusarium head blight in the main cropping area of Argentina were investigated in heavily infected samples. The ability of the isolates to produce trichothecenes was determined by GC and HPLC. Fusarium graminearum was the predominant species and of 33 isolates, 10 produced deoxinivalenol (DON) (0.1- 29 mg kg(-1)), 13 produced both deoxinivalenol (1.0- 708 mg kg(-1)) and nivalenol (0.1- 6.2mg kg(-1)), 12 produced 3-acetyldeoxinivalenol (0.1- 14 mg kg(-1)), 13 produced 15-acetyldeoxinivalenol (0.1- 1.9 mg kg(-1)), 10 produced Fusarenone X (0.1- 2.4 mg kg(-1)) and 7 produced zearalenone (0.1- 0.6 mg kg(-1)). These results suggest that F. graminearum strains isolated from the wheat growing regions in Argentina belong to DON chemotype. Although some strains produced both deoxinivalenol and nivalenol, nivalenol was produced in lower levels. The natural occurrence of nivalenol in wheat affected by head-blight collected in the main production area during two years (2001-2002) was also determined. From 19 samples 13 were contaminated with deoxinivalenol in a range of 0.3 to 70 mg kg(-1)and 2 samples with both deoxinivalenol (7.5 and 6.7 mg kg(-1)) and nivalenol (0.05 and 0.1 mg kg(-1)), respectively. This is the first report of natural occurrence of nivalenol in wheat cultivate in Argentina.

Natural occurrence of nivalenol and mycotoxigenic potential of Fusarium graminearum strains in wheat affected by head blight in Argentina

The principal agents of Fusarium head blight in the main cropping area of Argentina were investigated in heavily infected samples. The ability of the isolates to produce trichothecenes was determined by GC and HPLC. Fusarium graminearum was the predominant species and of 33 isolates, 10 produced deoxinivalenol (DON) (0.1- 29 mg kg-1), 13 produced both deoxinivalenol (1.0- 708 mg kg-1) and nivalenol (0.1- 6.2mg kg-1), 12 produced 3-acetyldeoxinivalenol (0.1- 14 mg kg-1), 13 produced 15-acetyldeoxinivalenol (0.1- 1.9 mg kg-1), 10 produced Fusarenone X (0.1- 2.4 mg kg-1) and 7 produced zearalenone (0.1- 0.6 mg kg-1). These results suggest that F. graminearum strains isolated from the wheat growing regions in Argentina belong to DON chemotype. Although some strains produced both deoxinivalenol and nivalenol, nivalenol was produced in lower levels. The natural occurrence of nivalenol in wheat affected by head-blight collected in the main production area during two years (2001-2002) was also determined. From 19 samples 13 were contaminated with deoxinivalenol in a range of 0.3 to 70 mg kg-1and 2 samples with both deoxinivalenol (7.5 and 6.7 mg kg-1) and nivalenol (0.05 and 0.1 mg kg-1), respectively. This is the first report of natural occurrence of nivalenol in wheat cultivate in Argentina.

O principal causador de giberela no trigo na Argentina e sua capacidade de produzir tricotecenos foram estudados por GC e HPLC em amostras altamente infectadas. A espécie predominante foi Fusarium graminearum, sendo que de um total de 33 isolados, 10 produziram deoxinivalenol (0,1-29 mg kg -1), 13 produziram deoxinivalenol (1,0-708 mg kg-1) e nivalenol (0,1-6,2 mg kg-1), 12 produziram 3-acetildeoxinivalenol (0,1-14 mg kg-1), 13 produziram 15-acetildeoxinivalenol (0,1-1,9 mg kg-1), 10 produziram fusarenona X (0,1- 2,4 mg kg-1) e 7 produziram zearalenona (0,1- 0,6 mg kg-1). Esses resultados sugerem que as cepas de F. graminearum isoladas de trigo cultivado na Argentina pertencem ao quimiotipo DON. Embora algumas cepas tenham produzido tanto DON quanto NIV, NIV foi produzido em quantidade inferior ao DON. A ocorrência natural de nivalenol em trigo afetado pela giberela coletado na principal área de produção durante dois anos (2001-2002) foi também determinada. De 19 amostras, 13 estavam contaminadas com deoxinivalenol na faixa de 0,3 a 70 mg kg-1 e 2amostras continham tanto deoxinivalenol (7,5 e 6,7 mg kg-1) quanto nivalenol (0,05 e 0,1 mg kg-1), respectivamente. Esse é o primeiro relato da ocorrência de nivalenol em trigo cultivado na Argentina.