Nikolić, Milica

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  • Nikolić, Milica (7)
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First report of Aspergillus parasiticus on Barley Grain in Serbia

Nikolić, Milica; Savić, Iva; Obradović, Ana; Srdić, Jelena; Stanković, Goran; Stevanović, Milan; Stanković, Slavica

(St. Paul : The American Phytopathological Society (APS), 2020)

TY  - JOUR
AU  - Nikolić, Milica
AU  - Savić, Iva
AU  - Obradović, Ana
AU  - Srdić, Jelena
AU  - Stanković, Goran
AU  - Stevanović, Milan
AU  - Stanković, Slavica
PY  - 2020
UR  - http://rik.mrizp.rs/handle/123456789/796
AB  - Barley (Hordeum vulgare L.) is a secondary grain crop in Serbia used commercially for animal feed, seed, and human food applications. The production of barley in the 2016 to 2017 growing season reached a record yield of almost 400,000 metric tons (USDA 2017). Aspergillus contamination has been rare in the agroecological conditions of cereal-growing areas in Serbia. Changes in climatic factors, such as occurrence of high temperatures and prolonged droughts, increased frequency of Aspergillus spp. Species Aspergillus parasiticus was isolated from maize grain for the first time in Serbia in 2012 and from wheat grains in 2017 (Nikolic et al. 2018). We hypothesized that these pathogens can also be present in barley fields in Serbia. Barley spikes exhibiting bleaching were sampled at the beginning of June 2017 grown in northern Serbia. In severe infections, barley spikes get a dry look with awns that stand upright and firm. The incidence of the disease of the bleached spiked in the field was 15 to 20%. From each representative sample, 100 shriveled grains were collected. After surface sterilization with bleach/distilled water 1:3, 100 grains per sample (10 per Petri dish) were placed on potato dextrose agar and incubated at 25°C for 7 days. After isolation, 50% of isolates were identified as Alternaria spp., 20% as Fusarium spp., approximately 15% as Aspergillus spp. In order to reliably identify individual species of fungi, the fragments of colonies were transferred to malt extract agar (MEA) and Czapek yeast agar (CYA) and incubated in the dark for 7 days. The fungal colonies were dark green. The reverse side was pale yellow. The average colony diameter was 65 mm. Conidia were spherical and rough with thick walls. The average size of conidia was 5.1 µm. Colonies were floccose and thin on MEA and CYA. Based on growth and morphological characteristics, isolates were determined as A. parasiticus (Pitt and Hocking 2009). Molecular detection of Aspergillus species was done by using PCR-restriction fragment length polymorphism analysis of the aflR-aflJ (genes for aflatoxin biosynthesis) intergenic spacer. The restriction enzyme BglII was able to cut the PCR product of A. parasiticus at one restriction site, resulting in two fragments of 363 and 311 bp (El Khoury et al. 2011). A. parasiticus CBS 100926 was used as a reference isolate. The pathogenicity of 20 isolates was verified on a group of 20 randomly selected spikes in four replicates (Mesterházy et al. 1999). A 7-day-old culture of each isolate was used for the preparation of the spore suspension (1 × 10⁶ spores/ml). Inoculation was carried out after 50% of plants reached the anthesis stage. Groups of 20 selected spikes were sprayed from all sides with 20 ml of fungal spore suspension. Control spikes were inoculated by applying an equal amount of sterile distilled water. The infection rate was estimated after 3 weeks on a 1 to 7 scale, with 1 = 0 to 5%, 2 = 5 to 15%, 3 = 15 to 30%, 4 = 30 to 50%, 5 = 50 to 75%, 6 = 75 to 90%, and 7 = 90 to 100% infected spike area. The average infection rate was 3.2. The pathogen was reisolated from the inoculated spikes and identified as A. parasiticus, with the aim to confirm Koch’s postulates. Developed symptoms were similar to those observed on spikes collected from the field. Control spikes did not show any symptoms of the disease. These results confirmed the pathogenicity of A. parasiticus on H. vulgare. To the best of our knowledge, this is the first report of the occurrence of A. parasiticus on barley grain in Serbia. Because A. parasiticus is known to be a severe aflatoxin producer and climatic changes can increase the frequency of this fungus, further studies are necessary to improve strategies for food safety and quality.
PB  - St. Paul : The American Phytopathological Society (APS)
T2  - Plant Disease
T1  - First report of Aspergillus parasiticus on Barley Grain in Serbia
VL  - 104
IS  - 3
SP  - 987
DO  - 10.1094/PDIS-07-19-1364-PDN
ER  - 
@article{
author = "Nikolić, Milica and Savić, Iva and Obradović, Ana and Srdić, Jelena and Stanković, Goran and Stevanović, Milan and Stanković, Slavica",
year = "2020",
url = "http://rik.mrizp.rs/handle/123456789/796",
abstract = "Barley (Hordeum vulgare L.) is a secondary grain crop in Serbia used commercially for animal feed, seed, and human food applications. The production of barley in the 2016 to 2017 growing season reached a record yield of almost 400,000 metric tons (USDA 2017). Aspergillus contamination has been rare in the agroecological conditions of cereal-growing areas in Serbia. Changes in climatic factors, such as occurrence of high temperatures and prolonged droughts, increased frequency of Aspergillus spp. Species Aspergillus parasiticus was isolated from maize grain for the first time in Serbia in 2012 and from wheat grains in 2017 (Nikolic et al. 2018). We hypothesized that these pathogens can also be present in barley fields in Serbia. Barley spikes exhibiting bleaching were sampled at the beginning of June 2017 grown in northern Serbia. In severe infections, barley spikes get a dry look with awns that stand upright and firm. The incidence of the disease of the bleached spiked in the field was 15 to 20%. From each representative sample, 100 shriveled grains were collected. After surface sterilization with bleach/distilled water 1:3, 100 grains per sample (10 per Petri dish) were placed on potato dextrose agar and incubated at 25°C for 7 days. After isolation, 50% of isolates were identified as Alternaria spp., 20% as Fusarium spp., approximately 15% as Aspergillus spp. In order to reliably identify individual species of fungi, the fragments of colonies were transferred to malt extract agar (MEA) and Czapek yeast agar (CYA) and incubated in the dark for 7 days. The fungal colonies were dark green. The reverse side was pale yellow. The average colony diameter was 65 mm. Conidia were spherical and rough with thick walls. The average size of conidia was 5.1 µm. Colonies were floccose and thin on MEA and CYA. Based on growth and morphological characteristics, isolates were determined as A. parasiticus (Pitt and Hocking 2009). Molecular detection of Aspergillus species was done by using PCR-restriction fragment length polymorphism analysis of the aflR-aflJ (genes for aflatoxin biosynthesis) intergenic spacer. The restriction enzyme BglII was able to cut the PCR product of A. parasiticus at one restriction site, resulting in two fragments of 363 and 311 bp (El Khoury et al. 2011). A. parasiticus CBS 100926 was used as a reference isolate. The pathogenicity of 20 isolates was verified on a group of 20 randomly selected spikes in four replicates (Mesterházy et al. 1999). A 7-day-old culture of each isolate was used for the preparation of the spore suspension (1 × 10⁶ spores/ml). Inoculation was carried out after 50% of plants reached the anthesis stage. Groups of 20 selected spikes were sprayed from all sides with 20 ml of fungal spore suspension. Control spikes were inoculated by applying an equal amount of sterile distilled water. The infection rate was estimated after 3 weeks on a 1 to 7 scale, with 1 = 0 to 5%, 2 = 5 to 15%, 3 = 15 to 30%, 4 = 30 to 50%, 5 = 50 to 75%, 6 = 75 to 90%, and 7 = 90 to 100% infected spike area. The average infection rate was 3.2. The pathogen was reisolated from the inoculated spikes and identified as A. parasiticus, with the aim to confirm Koch’s postulates. Developed symptoms were similar to those observed on spikes collected from the field. Control spikes did not show any symptoms of the disease. These results confirmed the pathogenicity of A. parasiticus on H. vulgare. To the best of our knowledge, this is the first report of the occurrence of A. parasiticus on barley grain in Serbia. Because A. parasiticus is known to be a severe aflatoxin producer and climatic changes can increase the frequency of this fungus, further studies are necessary to improve strategies for food safety and quality.",
publisher = "St. Paul : The American Phytopathological Society (APS)",
journal = "Plant Disease",
title = "First report of Aspergillus parasiticus on Barley Grain in Serbia",
volume = "104",
number = "3",
pages = "987",
doi = "10.1094/PDIS-07-19-1364-PDN"
}
Nikolić, M., Savić, I., Obradović, A., Srdić, J., Stanković, G., Stevanović, M.,& Stanković, S. (2020). First report of Aspergillus parasiticus on Barley Grain in Serbia.
Plant Disease
St. Paul : The American Phytopathological Society (APS)., 104(3), 987.
https://doi.org/10.1094/PDIS-07-19-1364-PDN
Nikolić M, Savić I, Obradović A, Srdić J, Stanković G, Stevanović M, Stanković S. First report of Aspergillus parasiticus on Barley Grain in Serbia. Plant Disease. 2020;104(3):987
Nikolić Milica, Savić Iva, Obradović Ana, Srdić Jelena, Stanković Goran, Stevanović Milan, Stanković Slavica, "First report of Aspergillus parasiticus on Barley Grain in Serbia" Plant Disease, 104, no. 3 (2020):987,
https://doi.org/10.1094/PDIS-07-19-1364-PDN .

Aspergillus parasiticus Speare – toksigena vrsta gljive na zrnu kukuruza u proizvodnim uslovima Srbije

Nikolić, Milica

(2019)

TY  - BOOK
AU  - Nikolić, Milica
PY  - 2019
UR  - http://rik.mrizp.rs/handle/123456789/771
AB  - In the course of four-year studies (2013-2016), the species Aspergillus parasiticus was identified in Serbia for the first time as a causal agent of aspergillus kernel and ear rot in both fields and storages. In order to examine the distribution and diversity of this species, the study of morphological, pathogenic, molecular and toxigenic properties of 48 isolates of this species selected from maize kernels was carried out. Studies of macroscopic morphological traits of isolates showed that biodiversity of observed isolates was low. Colonies of the isolates formed on PDA the abundant dark green, dense cottony mycelium coating. All isolates of A. parasiticus formed unicellular denticulate conidia on MEA. The diameter of conidia of observed isolates varied from 4.08μm to 6.86μm.Two factorial analysis of variance showed that the effects of interactions of studied isolates, light regime and temperatures (p<0.01) on daily growth of the colony diameters were statistically significant. The pathogenicity test pointed out that all observed isolates of A. parasiticus were pathogens of maize ears, as well as that there was divergence in the virulence expression.The comparison of concentrations of synthesised mycotoxins in isolates obtained by ELISA showed a statistically highly significant positive correlation between potential to produce AFB1 and AFLA in tested isolates (r=0,66 μm).The observed isolates A. parasiticus that synthesised aflatoxins (50%) mostly synthesised AFB1 (84.78%), while the number of isolates that synthesised AFG1 was significantly lower (15.22%). However, it was established that eight isolates synthesised AFG1 at higher concentrations than AFB1. Mycotoxin concentrations in A. parasiticusisolates were compared by using the results obtained by the HPLC method. The comparison indicated that there was a highly statistically positive correlation between synthesised concentrations of AFB1 and AFG1 (r=0.82 μm), as well as between AFB2 and AFG2 (r=0.63 μm).The identity of all isolates previously characterised on the basis of morphological, ecological and toxigenic properties as species A. parasiticus was confirmed by the multiplex PCR detection method. Distinguishing between species A. parasiticus and morphologically similar species A. flavus was enabled by the application of the RFLP-PCR method, using restriction enzymes. This was also the first identification of this pathogenic species at the molecular level in Serbia.
AB  - U toku četvorogodišnjih ispitivanja (2013-2016) identifikovana je, po prvi put u Srbiji, vrsta Asperigillus parasiticus kao prouzrokovač aspergilozne truleži zrna i klipa kukuruza u polju i skladištima. Sa ciljem da se ispita rasprostranjenost i diverzitet ove vrste obavljena su proučavanja morfoloških, patogenih, molekularnih, odgajivačkih i toksigenih svojstava 46 odabranih izolata ove vrste poreklom sa zrna kukuruza. Ispitivanjem makroskopskih morfoloških karakteristika izolata ukazano je na mali biodiverzitet proučavanih izolata. Kolonije izolata na PDA hranljivoj podlozi formirale su obilnu tamnozelenu, gustu prevlaku od micelije gljive, pamučastog izgleda. Svi izolati A. parasiticus obrazuju jednoćelijske, nazubljene konidije na MEA hranljivoj podlozi. Prečnik konidija ispitivanih izolata varirao je od 4,08μm do 6,86μm. Dvofaktorska analiza varijanse pokazala je statistički značajan uticaj interakcije ispitivanih izolata, svetlosnog režima i temperature (p<0.01) na dnevni porast prečnika kolonija. Test patogenosti ukazao je da su svi ispitivani izolati A. parasiticus patogeni na klipu kukuruza, kao i da postoji divergentnost u ispoljavanju virulentnosti. Poređenjem koncentracija sintetisanih mikotoksina kod izolata dobijenih Elisa testom, pokazali su da postoji statistički visoko značajna pozitivna korelacija između potencijala produkcije AFB1 i AFLA kod ispitivanih izolata (r=0,66 μm). Ispitivani izolati A.parasiticus koji su sintetisali aflatoksine (50%) su u najvećem procentu sintetisali AFB1 (84,78%), dok je procenat izolata koji su sintetisali AFG1 bila znatno manja (15,22%). Međutim, utvrđeno je osam izolata koji su sintetisali AFG1 u višim koncentracijama, u odnosu na AFB1. Poređenjem koncentracija mikotoksina kod izolata A. parasiticus, na osnovu rezultata HPLC metode, uočeno je da postoji visoko statistički značajna pozitivna korelacija između sintetisanih koncentracija AFB1 i AFG1 (r=0,82 μm), kao i između AFB2 i AFG2 (r=0,63 μm). Multiplex PCR detekcijom potvrđen je identitet svih izolata prethodno okarakterisanih na osnovu morfoloških, ekoloških, odgajivačkih i toksigenih karakteristika kao vrsta A. parasiticus. Primenom RFLP-PCR metode korišćenjem restrikcionih enzima, omogućeno je razlikovanje vrste A. parasiticus od morfološki slične vrste A. flavus, što je ujedno i prva identifikacija ove patogene vrste na molekularnom nivou u Srbiji.
T1  - Aspergillus parasiticus Speare – toksigena vrsta gljive na zrnu kukuruza u proizvodnim uslovima Srbije
T1  - Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia
ER  - 
@book{
author = "Nikolić, Milica",
year = "2019",
url = "http://rik.mrizp.rs/handle/123456789/771",
abstract = "In the course of four-year studies (2013-2016), the species Aspergillus parasiticus was identified in Serbia for the first time as a causal agent of aspergillus kernel and ear rot in both fields and storages. In order to examine the distribution and diversity of this species, the study of morphological, pathogenic, molecular and toxigenic properties of 48 isolates of this species selected from maize kernels was carried out. Studies of macroscopic morphological traits of isolates showed that biodiversity of observed isolates was low. Colonies of the isolates formed on PDA the abundant dark green, dense cottony mycelium coating. All isolates of A. parasiticus formed unicellular denticulate conidia on MEA. The diameter of conidia of observed isolates varied from 4.08μm to 6.86μm.Two factorial analysis of variance showed that the effects of interactions of studied isolates, light regime and temperatures (p<0.01) on daily growth of the colony diameters were statistically significant. The pathogenicity test pointed out that all observed isolates of A. parasiticus were pathogens of maize ears, as well as that there was divergence in the virulence expression.The comparison of concentrations of synthesised mycotoxins in isolates obtained by ELISA showed a statistically highly significant positive correlation between potential to produce AFB1 and AFLA in tested isolates (r=0,66 μm).The observed isolates A. parasiticus that synthesised aflatoxins (50%) mostly synthesised AFB1 (84.78%), while the number of isolates that synthesised AFG1 was significantly lower (15.22%). However, it was established that eight isolates synthesised AFG1 at higher concentrations than AFB1. Mycotoxin concentrations in A. parasiticusisolates were compared by using the results obtained by the HPLC method. The comparison indicated that there was a highly statistically positive correlation between synthesised concentrations of AFB1 and AFG1 (r=0.82 μm), as well as between AFB2 and AFG2 (r=0.63 μm).The identity of all isolates previously characterised on the basis of morphological, ecological and toxigenic properties as species A. parasiticus was confirmed by the multiplex PCR detection method. Distinguishing between species A. parasiticus and morphologically similar species A. flavus was enabled by the application of the RFLP-PCR method, using restriction enzymes. This was also the first identification of this pathogenic species at the molecular level in Serbia., U toku četvorogodišnjih ispitivanja (2013-2016) identifikovana je, po prvi put u Srbiji, vrsta Asperigillus parasiticus kao prouzrokovač aspergilozne truleži zrna i klipa kukuruza u polju i skladištima. Sa ciljem da se ispita rasprostranjenost i diverzitet ove vrste obavljena su proučavanja morfoloških, patogenih, molekularnih, odgajivačkih i toksigenih svojstava 46 odabranih izolata ove vrste poreklom sa zrna kukuruza. Ispitivanjem makroskopskih morfoloških karakteristika izolata ukazano je na mali biodiverzitet proučavanih izolata. Kolonije izolata na PDA hranljivoj podlozi formirale su obilnu tamnozelenu, gustu prevlaku od micelije gljive, pamučastog izgleda. Svi izolati A. parasiticus obrazuju jednoćelijske, nazubljene konidije na MEA hranljivoj podlozi. Prečnik konidija ispitivanih izolata varirao je od 4,08μm do 6,86μm. Dvofaktorska analiza varijanse pokazala je statistički značajan uticaj interakcije ispitivanih izolata, svetlosnog režima i temperature (p<0.01) na dnevni porast prečnika kolonija. Test patogenosti ukazao je da su svi ispitivani izolati A. parasiticus patogeni na klipu kukuruza, kao i da postoji divergentnost u ispoljavanju virulentnosti. Poređenjem koncentracija sintetisanih mikotoksina kod izolata dobijenih Elisa testom, pokazali su da postoji statistički visoko značajna pozitivna korelacija između potencijala produkcije AFB1 i AFLA kod ispitivanih izolata (r=0,66 μm). Ispitivani izolati A.parasiticus koji su sintetisali aflatoksine (50%) su u najvećem procentu sintetisali AFB1 (84,78%), dok je procenat izolata koji su sintetisali AFG1 bila znatno manja (15,22%). Međutim, utvrđeno je osam izolata koji su sintetisali AFG1 u višim koncentracijama, u odnosu na AFB1. Poređenjem koncentracija mikotoksina kod izolata A. parasiticus, na osnovu rezultata HPLC metode, uočeno je da postoji visoko statistički značajna pozitivna korelacija između sintetisanih koncentracija AFB1 i AFG1 (r=0,82 μm), kao i između AFB2 i AFG2 (r=0,63 μm). Multiplex PCR detekcijom potvrđen je identitet svih izolata prethodno okarakterisanih na osnovu morfoloških, ekoloških, odgajivačkih i toksigenih karakteristika kao vrsta A. parasiticus. Primenom RFLP-PCR metode korišćenjem restrikcionih enzima, omogućeno je razlikovanje vrste A. parasiticus od morfološki slične vrste A. flavus, što je ujedno i prva identifikacija ove patogene vrste na molekularnom nivou u Srbiji.",
title = "Aspergillus parasiticus Speare – toksigena vrsta gljive na zrnu kukuruza u proizvodnim uslovima Srbije, Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia"
}
Nikolić, M. (2019). Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia.
.
Nikolić M. Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia. 2019;
Nikolić Milica, "Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia" (2019)

Aspergillus parasiticus Speare – toksigena vrsta gljive na zrnu kukuruza u proizvodnim uslovima Srbije

Nikolić, Milica

(Novi Sad, Univerzitet u Novom Sadu, Poljoprivredni fakultet, 2019)

TY  - BOOK
AU  - Nikolić, Milica
PY  - 2019
UR  - http://rik.mrizp.rs/handle/123456789/770
AB  - In the course of four-year studies (2013-2016), the species Aspergillus parasiticus was identified in Serbia for the first time as a causal agent of aspergillus kernel and ear rot in both fields and storages. In order to examine the distribution and diversity of this species, the study of morphological, pathogenic, molecular and toxigenic properties of 48 isolates of this species selected from maize kernels was carried out. Studies of macroscopic morphological traits of isolates showed that biodiversity of observed isolates was low. Colonies of the isolates formed on PDA the abundant dark green, dense cottony mycelium coating. All isolates of A. parasiticus formed unicellular denticulate conidia on MEA. The diameter of conidia of observed isolates varied from 4.08μm to 6.86μm.
Two factorial analysis of variance showed that the effects of interactions of studied isolates, light regime and temperatures (p<0.01) on daily growth of the colony diameters were statistically significant. The pathogenicity test pointed out that all observed isolates of A. parasiticus were pathogens of maize ears, as well as that there was divergence in the virulence expression.The comparison of concentrations of synthesised mycotoxins in isolates obtained by ELISA showed a statistically highly significant positive correlation between potential to produce AFB1 and AFLA in tested isolates (r=0,66 μm).The observed isolates A. parasiticus that synthesised aflatoxins (50%) mostly synthesised AFB1 (84.78%), while the number of isolates that synthesised AFG1 was significantly lower (15.22%). However, it was established that eight isolates synthesised AFG1 at higher concentrations than AFB1. Mycotoxin concentrations in A. parasiticusisolates were compared by using the results obtained by the HPLC method. The comparison indicated that there was a highly statistically positive correlation between synthesised concentrations of AFB1 and AFG1 (r=0.82 μm), as well as between AFB2 and AFG2 (r=0.63 μm).The identity of all isolates previously characterised on the basis of morphological, ecological and toxigenic properties as species A. parasiticus was confirmed by the multiplex PCR detection method. Distinguishing between species A. parasiticus and morphologically similar species A. flavus was enabled by the application of the RFLP-PCR method, using restriction enzymes. This was also the first identification of this pathogenic species at the molecular level in Serbia.
AB  - U toku četvorogodišnjih ispitivanja (2013-2016) identifikovana je, po prvi put u Srbiji, vrsta Asperigillus parasiticus kao prouzrokovač aspergilozne truleži zrna i klipa kukuruza u polju i skladištima. Sa ciljem da se ispita rasprostranjenost i diverzitet ove vrste obavljena su proučavanja morfoloških, patogenih, molekularnih, odgajivačkih i toksigenih svojstava 46 odabranih izolata ove vrste poreklom sa zrna kukuruza. Ispitivanjem makroskopskih morfoloških karakteristika izolata ukazano je na mali biodiverzitet proučavanih izolata. Kolonije izolata na PDA hranljivoj podlozi formirale su obilnu tamnozelenu, gustu prevlaku od micelije gljive, pamučastog izgleda. Svi izolati A. parasiticus obrazuju jednoćelijske, nazubljene konidije na MEA hranljivoj podlozi. Prečnik konidija ispitivanih izolata varirao je od 4,08μm do 6,86μm. Dvofaktorska analiza varijanse pokazala je statistički značajan uticaj interakcije ispitivanih izolata, svetlosnog režima i temperature (p<0.01) na dnevni porast prečnika kolonija. Test patogenosti ukazao je da su svi ispitivani izolati A. parasiticus patogeni na klipu kukuruza, kao i da postoji divergentnost u ispoljavanju virulentnosti. Poređenjem koncentracija sintetisanih mikotoksina kod izolata dobijenih Elisa testom, pokazali su da postoji statistički visoko značajna pozitivna korelacija između potencijala produkcije AFB1 i AFLA kod ispitivanih izolata (r=0,66 μm). Ispitivani izolati A.parasiticus koji su sintetisali aflatoksine (50%) su u najvećem procentu sintetisali AFB1 (84,78%), dok je procenat izolata koji su sintetisali AFG1 bila znatno manja (15,22%). Međutim, utvrđeno je osam izolata koji su sintetisali AFG1 u višim koncentracijama, u odnosu na AFB1. Poređenjem koncentracija mikotoksina kod izolata A. parasiticus, na osnovu rezultata HPLC metode, uočeno je da postoji visoko statistički značajna pozitivna korelacija između sintetisanih koncentracija AFB1 i AFG1 (r=0,82 μm), kao i između AFB2 i AFG2 (r=0,63 μm). Multiplex PCR detekcijom potvrđen je identitet svih izolata prethodno okarakterisanih na osnovu morfoloških, ekoloških, odgajivačkih i toksigenih karakteristika kao vrsta A. parasiticus. Primenom RFLP-PCR metode korišćenjem restrikcionih enzima, omogućeno je razlikovanje vrste A. parasiticus od morfološki slične vrste A. flavus, što je ujedno i prva identifikacija ove patogene vrste na molekularnom nivou u Srbiji.
PB  - Novi Sad, Univerzitet u Novom Sadu, Poljoprivredni fakultet
T1  - Aspergillus parasiticus Speare – toksigena vrsta gljive na zrnu kukuruza u proizvodnim uslovima Srbije
T1  - Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia
ER  - 
@phdthesis{
author = "Nikolić, Milica",
year = "2019",
url = "http://rik.mrizp.rs/handle/123456789/770",
abstract = "In the course of four-year studies (2013-2016), the species Aspergillus parasiticus was identified in Serbia for the first time as a causal agent of aspergillus kernel and ear rot in both fields and storages. In order to examine the distribution and diversity of this species, the study of morphological, pathogenic, molecular and toxigenic properties of 48 isolates of this species selected from maize kernels was carried out. Studies of macroscopic morphological traits of isolates showed that biodiversity of observed isolates was low. Colonies of the isolates formed on PDA the abundant dark green, dense cottony mycelium coating. All isolates of A. parasiticus formed unicellular denticulate conidia on MEA. The diameter of conidia of observed isolates varied from 4.08μm to 6.86μm.
Two factorial analysis of variance showed that the effects of interactions of studied isolates, light regime and temperatures (p<0.01) on daily growth of the colony diameters were statistically significant. The pathogenicity test pointed out that all observed isolates of A. parasiticus were pathogens of maize ears, as well as that there was divergence in the virulence expression.The comparison of concentrations of synthesised mycotoxins in isolates obtained by ELISA showed a statistically highly significant positive correlation between potential to produce AFB1 and AFLA in tested isolates (r=0,66 μm).The observed isolates A. parasiticus that synthesised aflatoxins (50%) mostly synthesised AFB1 (84.78%), while the number of isolates that synthesised AFG1 was significantly lower (15.22%). However, it was established that eight isolates synthesised AFG1 at higher concentrations than AFB1. Mycotoxin concentrations in A. parasiticusisolates were compared by using the results obtained by the HPLC method. The comparison indicated that there was a highly statistically positive correlation between synthesised concentrations of AFB1 and AFG1 (r=0.82 μm), as well as between AFB2 and AFG2 (r=0.63 μm).The identity of all isolates previously characterised on the basis of morphological, ecological and toxigenic properties as species A. parasiticus was confirmed by the multiplex PCR detection method. Distinguishing between species A. parasiticus and morphologically similar species A. flavus was enabled by the application of the RFLP-PCR method, using restriction enzymes. This was also the first identification of this pathogenic species at the molecular level in Serbia., U toku četvorogodišnjih ispitivanja (2013-2016) identifikovana je, po prvi put u Srbiji, vrsta Asperigillus parasiticus kao prouzrokovač aspergilozne truleži zrna i klipa kukuruza u polju i skladištima. Sa ciljem da se ispita rasprostranjenost i diverzitet ove vrste obavljena su proučavanja morfoloških, patogenih, molekularnih, odgajivačkih i toksigenih svojstava 46 odabranih izolata ove vrste poreklom sa zrna kukuruza. Ispitivanjem makroskopskih morfoloških karakteristika izolata ukazano je na mali biodiverzitet proučavanih izolata. Kolonije izolata na PDA hranljivoj podlozi formirale su obilnu tamnozelenu, gustu prevlaku od micelije gljive, pamučastog izgleda. Svi izolati A. parasiticus obrazuju jednoćelijske, nazubljene konidije na MEA hranljivoj podlozi. Prečnik konidija ispitivanih izolata varirao je od 4,08μm do 6,86μm. Dvofaktorska analiza varijanse pokazala je statistički značajan uticaj interakcije ispitivanih izolata, svetlosnog režima i temperature (p<0.01) na dnevni porast prečnika kolonija. Test patogenosti ukazao je da su svi ispitivani izolati A. parasiticus patogeni na klipu kukuruza, kao i da postoji divergentnost u ispoljavanju virulentnosti. Poređenjem koncentracija sintetisanih mikotoksina kod izolata dobijenih Elisa testom, pokazali su da postoji statistički visoko značajna pozitivna korelacija između potencijala produkcije AFB1 i AFLA kod ispitivanih izolata (r=0,66 μm). Ispitivani izolati A.parasiticus koji su sintetisali aflatoksine (50%) su u najvećem procentu sintetisali AFB1 (84,78%), dok je procenat izolata koji su sintetisali AFG1 bila znatno manja (15,22%). Međutim, utvrđeno je osam izolata koji su sintetisali AFG1 u višim koncentracijama, u odnosu na AFB1. Poređenjem koncentracija mikotoksina kod izolata A. parasiticus, na osnovu rezultata HPLC metode, uočeno je da postoji visoko statistički značajna pozitivna korelacija između sintetisanih koncentracija AFB1 i AFG1 (r=0,82 μm), kao i između AFB2 i AFG2 (r=0,63 μm). Multiplex PCR detekcijom potvrđen je identitet svih izolata prethodno okarakterisanih na osnovu morfoloških, ekoloških, odgajivačkih i toksigenih karakteristika kao vrsta A. parasiticus. Primenom RFLP-PCR metode korišćenjem restrikcionih enzima, omogućeno je razlikovanje vrste A. parasiticus od morfološki slične vrste A. flavus, što je ujedno i prva identifikacija ove patogene vrste na molekularnom nivou u Srbiji.",
publisher = "Novi Sad, Univerzitet u Novom Sadu, Poljoprivredni fakultet",
title = "Aspergillus parasiticus Speare – toksigena vrsta gljive na zrnu kukuruza u proizvodnim uslovima Srbije, Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia"
}
Nikolić, M. (2019). Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia.

Novi Sad, Univerzitet u Novom Sadu, Poljoprivredni fakultet..
Nikolić M. Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia. 2019;
Nikolić Milica, "Aspergillus parasiticusSpeare –a toxigenic species on maize kernels underproduction conditions in Serbia" (2019)

Impacts of climatic conditions on aflatoxin B 1 and fumonisins contamination of maize kernels and their co-occurrence

Obradović, Ana; Krnjaja, Vesna; Nikolić, Milica; Delibašić, Goran; Filipović, Milomir; Stanković, Goran; Stanković, Slavica

(Institut za stočarstvo, Beograd, 2018)

TY  - JOUR
AU  - Obradović, Ana
AU  - Krnjaja, Vesna
AU  - Nikolić, Milica
AU  - Delibašić, Goran
AU  - Filipović, Milomir
AU  - Stanković, Goran
AU  - Stanković, Slavica
PY  - 2018
UR  - http://rik.mrizp.rs/handle/123456789/738
AB  - Agroecological and climatic conditions in Serbia greatly affected the development of toxigenic fungi and occurrence of mycotoxins in the maize. The presence of fungal toxigenic Aspergillus and Fusarium species and levels of aflatoxin B 1 (AFB 1) and sum of fumonisins B 1 , B 2 and B 3 (FBs) were established in 127 maize kernel samples harvested during 2012 (37 samples) and 2013 (90 samples). The periods of silking and grain filling of the maize in 2012 in comparison to 2013 were characterised with extremely dry spells, with high temperatures and low precipitation sums. The mean incidences of A. flavus and F. verticillioides were 50.4 and 11.7% in 2012 and 18.9 and 33.4% in 2013, respectively. According to the regulations of the World Health Organisation, unacceptable levels of AFB 1 (>20 μg kg-1) and FBs (>2000 μg kg-1) were established in the 30.6 and 24.1% samples of 2012 and 16.7 and 40% maize kernel samples of 2013, respectively. It can be concluded that high temperatures and low precipitation sums in 2012 favoured the development of A. flavus affecting the high level of AFB 1 , in comparison with F. verticillioides and the production of FBs. There was no positive correlation between the incidences of A. flavus and F. verticillioides, while a statistically significant positive correlation has been found between AFB 1 and FBs levels, in both investigated years (2012-2013). This indicates that the mycotoxin production depended more on weather conditions than on the distribution of corresponding toxigenic fungal species.
AB  - Agroekološki i klimatski uslovi u Srbiji veoma su pogodni za razvoj toksigenih gljiva i njihovih mikotoksina u kukuruzu. Prisustvo toksigenih vrsta gljiva iz rodova Aspergillus i Fusarium, kao i nivoi aflatoksina B1 (AFB1) i ukupnih fumonizina B1, B2 i B3 (FBs) utvrđeni su u 127 uzoraka zrna kukuruza iz berbe tokom 2012 (37 uzoraka) i 2013. godine (90 uzoraka). U fenofazama svilanja i nalivanja zrna kukuruza u 2012. u odnosu na 2013. godinu zapaženi su ekstremno sušni periodi sa visokim temperaturama i niskim količinama padavina. Prosečne vrednosti za učestalost vrsta A. flavus i F. verticillioides bile su 50,4 i 11,7% u 2012., odnosno 18,9 i 33,4% u 2013. godini. Prema pravilniku Svetske zdravstvene organizacije (WHO) nedozvoljeni nivoi AFB1 (>20 µg/kg) i FBs (>2000 µg/kg) utvrđeni su u 30,6% i 24,1% uzoraka u 2012., odnosno u 16,7% i 40% uzoraka u 2013. godini. Na osnovu dobijenih rezultata zaključeno je da su visoke temperature i niske količine padavina u 2012. godini uslovile značajno veću učestalost A. flavus i visoku produkciju AFB1 u poredjenju sa vrstom F. verticillioides i produkcijom FBs. U obe ispitivane godine (2012-2013), između učestalosti A. flavus i F. verticillioides nije ustanovljena pozitivna korelacija, dok je statistički značajna pozitivna korelacija ustanovljena između nivoa AFB1 i FBs. Ovo ukazuje da produkcija mikotoksina je više zavisna od vremenskih uslova u odnosu na distribuciju toksigenih vrsta gljiva.
PB  - Institut za stočarstvo, Beograd
T2  - Biotechnology in Animal Husbandry
T1  - Impacts of climatic conditions on aflatoxin B 1 and fumonisins contamination of maize kernels and their co-occurrence
T1  - Uticaj klimatskih uslova na kontaminaciju zrna kukuruza sa aflatoksinom B1 i fumonizinima i njihova združena pojava
VL  - 34
IS  - 4
SP  - 469
EP  - 480
DO  - 10.2298/BAH1804469O
ER  - 
@article{
author = "Obradović, Ana and Krnjaja, Vesna and Nikolić, Milica and Delibašić, Goran and Filipović, Milomir and Stanković, Goran and Stanković, Slavica",
year = "2018",
url = "http://rik.mrizp.rs/handle/123456789/738",
abstract = "Agroecological and climatic conditions in Serbia greatly affected the development of toxigenic fungi and occurrence of mycotoxins in the maize. The presence of fungal toxigenic Aspergillus and Fusarium species and levels of aflatoxin B 1 (AFB 1) and sum of fumonisins B 1 , B 2 and B 3 (FBs) were established in 127 maize kernel samples harvested during 2012 (37 samples) and 2013 (90 samples). The periods of silking and grain filling of the maize in 2012 in comparison to 2013 were characterised with extremely dry spells, with high temperatures and low precipitation sums. The mean incidences of A. flavus and F. verticillioides were 50.4 and 11.7% in 2012 and 18.9 and 33.4% in 2013, respectively. According to the regulations of the World Health Organisation, unacceptable levels of AFB 1 (>20 μg kg-1) and FBs (>2000 μg kg-1) were established in the 30.6 and 24.1% samples of 2012 and 16.7 and 40% maize kernel samples of 2013, respectively. It can be concluded that high temperatures and low precipitation sums in 2012 favoured the development of A. flavus affecting the high level of AFB 1 , in comparison with F. verticillioides and the production of FBs. There was no positive correlation between the incidences of A. flavus and F. verticillioides, while a statistically significant positive correlation has been found between AFB 1 and FBs levels, in both investigated years (2012-2013). This indicates that the mycotoxin production depended more on weather conditions than on the distribution of corresponding toxigenic fungal species., Agroekološki i klimatski uslovi u Srbiji veoma su pogodni za razvoj toksigenih gljiva i njihovih mikotoksina u kukuruzu. Prisustvo toksigenih vrsta gljiva iz rodova Aspergillus i Fusarium, kao i nivoi aflatoksina B1 (AFB1) i ukupnih fumonizina B1, B2 i B3 (FBs) utvrđeni su u 127 uzoraka zrna kukuruza iz berbe tokom 2012 (37 uzoraka) i 2013. godine (90 uzoraka). U fenofazama svilanja i nalivanja zrna kukuruza u 2012. u odnosu na 2013. godinu zapaženi su ekstremno sušni periodi sa visokim temperaturama i niskim količinama padavina. Prosečne vrednosti za učestalost vrsta A. flavus i F. verticillioides bile su 50,4 i 11,7% u 2012., odnosno 18,9 i 33,4% u 2013. godini. Prema pravilniku Svetske zdravstvene organizacije (WHO) nedozvoljeni nivoi AFB1 (>20 µg/kg) i FBs (>2000 µg/kg) utvrđeni su u 30,6% i 24,1% uzoraka u 2012., odnosno u 16,7% i 40% uzoraka u 2013. godini. Na osnovu dobijenih rezultata zaključeno je da su visoke temperature i niske količine padavina u 2012. godini uslovile značajno veću učestalost A. flavus i visoku produkciju AFB1 u poredjenju sa vrstom F. verticillioides i produkcijom FBs. U obe ispitivane godine (2012-2013), između učestalosti A. flavus i F. verticillioides nije ustanovljena pozitivna korelacija, dok je statistički značajna pozitivna korelacija ustanovljena između nivoa AFB1 i FBs. Ovo ukazuje da produkcija mikotoksina je više zavisna od vremenskih uslova u odnosu na distribuciju toksigenih vrsta gljiva.",
publisher = "Institut za stočarstvo, Beograd",
journal = "Biotechnology in Animal Husbandry",
title = "Impacts of climatic conditions on aflatoxin B 1 and fumonisins contamination of maize kernels and their co-occurrence, Uticaj klimatskih uslova na kontaminaciju zrna kukuruza sa aflatoksinom B1 i fumonizinima i njihova združena pojava",
volume = "34",
number = "4",
pages = "469-480",
doi = "10.2298/BAH1804469O"
}
Obradović, A., Krnjaja, V., Nikolić, M., Delibašić, G., Filipović, M., Stanković, G.,& Stanković, S. (2018). Uticaj klimatskih uslova na kontaminaciju zrna kukuruza sa aflatoksinom B1 i fumonizinima i njihova združena pojava.
Biotechnology in Animal Husbandry
Institut za stočarstvo, Beograd., 34(4), 469-480.
https://doi.org/10.2298/BAH1804469O
Obradović A, Krnjaja V, Nikolić M, Delibašić G, Filipović M, Stanković G, Stanković S. Uticaj klimatskih uslova na kontaminaciju zrna kukuruza sa aflatoksinom B1 i fumonizinima i njihova združena pojava. Biotechnology in Animal Husbandry. 2018;34(4):469-480
Obradović Ana, Krnjaja Vesna, Nikolić Milica, Delibašić Goran, Filipović Milomir, Stanković Goran, Stanković Slavica, "Uticaj klimatskih uslova na kontaminaciju zrna kukuruza sa aflatoksinom B1 i fumonizinima i njihova združena pojava" Biotechnology in Animal Husbandry, 34, no. 4 (2018):469-480,
https://doi.org/10.2298/BAH1804469O .
4

Differentiation between aspergillus flavus and aspergillus parasiticus isolates originated from wheat

Nikolić, Milica; Nikolić, Ana; Savić, Iva; Petrović, Tanja; Stanković, Slavica; Jauković, Marko; Bagi, Ferenc

(Društvo genetičara Srbije, Beograd, 2018)

TY  - JOUR
AU  - Nikolić, Milica
AU  - Nikolić, Ana
AU  - Savić, Iva
AU  - Petrović, Tanja
AU  - Stanković, Slavica
AU  - Jauković, Marko
AU  - Bagi, Ferenc
PY  - 2018
UR  - http://rik.mrizp.rs/handle/123456789/709
AB  - The species of the genus Aspergillus, A. flavus and A. parasiticus, are the most aflatoxin-producing fungi. All previous studies carried out under the production conditions of Serbia showed no presence of A. parasiticus on wheat kernel. On the basis of changes in climatic factors, such as occurrence of high temperatures and prolonged droughts, which favour increased frequency of Aspergillus spp., we assumed that this pathogen can also be present in Serbia. The significance of direct losses as a consequence of wheat kernel infection, as well as potential contamination with aflatoxins, have pointed out to the need to determine the presence of toxigenic potential of A. flavus and A. parasiticus isolates originating from Serbia. For that purpose, wheat kernel samples were collected in nine locations. According to morphological, toxicological and molecular traits of isolated fungi, the presence of A. flavus and A. parasiticus was confirmed. This is the first time that A. parasiticus was identified on wheat under climatic conditions in Serbia. This study indicates that these pathogens may be a potential danger in wheat production in the region of Serbia. This danger will be much more certain if global climatic changes continue as they will provide more intensive development of these pathogens.
PB  - Društvo genetičara Srbije, Beograd
T2  - Genetika
T1  - Differentiation between aspergillus flavus and aspergillus parasiticus isolates originated from wheat
VL  - 50
IS  - 1
SP  - 143
EP  - 152
DO  - 10.2298/GENSR1801143N
ER  - 
@article{
author = "Nikolić, Milica and Nikolić, Ana and Savić, Iva and Petrović, Tanja and Stanković, Slavica and Jauković, Marko and Bagi, Ferenc",
year = "2018",
url = "http://rik.mrizp.rs/handle/123456789/709",
abstract = "The species of the genus Aspergillus, A. flavus and A. parasiticus, are the most aflatoxin-producing fungi. All previous studies carried out under the production conditions of Serbia showed no presence of A. parasiticus on wheat kernel. On the basis of changes in climatic factors, such as occurrence of high temperatures and prolonged droughts, which favour increased frequency of Aspergillus spp., we assumed that this pathogen can also be present in Serbia. The significance of direct losses as a consequence of wheat kernel infection, as well as potential contamination with aflatoxins, have pointed out to the need to determine the presence of toxigenic potential of A. flavus and A. parasiticus isolates originating from Serbia. For that purpose, wheat kernel samples were collected in nine locations. According to morphological, toxicological and molecular traits of isolated fungi, the presence of A. flavus and A. parasiticus was confirmed. This is the first time that A. parasiticus was identified on wheat under climatic conditions in Serbia. This study indicates that these pathogens may be a potential danger in wheat production in the region of Serbia. This danger will be much more certain if global climatic changes continue as they will provide more intensive development of these pathogens.",
publisher = "Društvo genetičara Srbije, Beograd",
journal = "Genetika",
title = "Differentiation between aspergillus flavus and aspergillus parasiticus isolates originated from wheat",
volume = "50",
number = "1",
pages = "143-152",
doi = "10.2298/GENSR1801143N"
}
Nikolić, M., Nikolić, A., Savić, I., Petrović, T., Stanković, S., Jauković, M.,& Bagi, F. (2018). Differentiation between aspergillus flavus and aspergillus parasiticus isolates originated from wheat.
Genetika
Društvo genetičara Srbije, Beograd., 50(1), 143-152.
https://doi.org/10.2298/GENSR1801143N
Nikolić M, Nikolić A, Savić I, Petrović T, Stanković S, Jauković M, Bagi F. Differentiation between aspergillus flavus and aspergillus parasiticus isolates originated from wheat. Genetika. 2018;50(1):143-152
Nikolić Milica, Nikolić Ana, Savić Iva, Petrović Tanja, Stanković Slavica, Jauković Marko, Bagi Ferenc, "Differentiation between aspergillus flavus and aspergillus parasiticus isolates originated from wheat" Genetika, 50, no. 1 (2018):143-152,
https://doi.org/10.2298/GENSR1801143N .
2
3
2

Comparison of methods for determination of the toxigenic potential of Aspergillus parasiticus Sp. and Aspergillus flavus L. isolated from maize

Nikolić, Milica; Stanković, Slavica; Savić, Iva

(Matica srpska, Novi Sad, 2017)

TY  - JOUR
AU  - Nikolić, Milica
AU  - Stanković, Slavica
AU  - Savić, Iva
PY  - 2017
UR  - http://rik.mrizp.rs/handle/123456789/666
AB  - Maize is considered one of the most susceptible crops to mycotoxins worldwide. Compared to other mycotoxins, the greatest attention has been paid to aflatoxins, due to their potential carcinogenicity and due to significant and longstanding problems they can cause in humans and animals. A. flavus and A. parasiticus produce aflatoxins in many economically significant crops in both fields and storages. Because of the potential aflatoxin contamination of maize grain, the toxigenic potential of A. flavus and A. parasiticus isolates, originating from Serbia, was tested in the present study. Furthermore, various applied methods for detection of these mycotoxins were compared in the study. Cultural, serological and analytical methods for the detection of mycotoxins were compared in the course of the experiment by the direct extraction of aflatoxins from the nutrient medium. The cultural methods for the detection of aflatoxin production were applied to 20 isolates of A. flavus (MRIZP Af18-20) and A. parasiticus (MRIZP Ap1-17). These methods are based on the yellow pigment formation in mycelia and nutrition media, occurrence of fluorescence on PDA (potato dextrose agar), agar containing β-cyclodextrine (CD-PDA), as well as on the red pigment formation after adding ammonium hydroxide to the existing medium. The ELISA was used to check quantitative and qualitative analyses of total aflatoxins (B1, B2, G1, G2) while the HPLC method was applied to establish ability of isolates to synthesize aflatoxins B1, B2, G1, G2. The yellow pigment formation, fluorescence and colony color changes of isolates into red, as a proof of toxigenicity of isolates, were confirmed in all cases by ELISA. A high potential of total aflatoxin production was determined in the majority of observed isolates. The ability of A. parasiticus isolates to synthesize aflatoxins G1 and G2 was confirmed by the HPLC method. This was essential for a better understanding of the key role of the suitability of cultural methods for preliminary evaluation of a large number of isolates. Our goal was to employ rapid biochemical approaches to prevent aflatoxin contamination of crops, and to reduce human and animal exposure to foodborne mycotoxins.
AB  - Kukuruz se širom sveta smatra jednim od useva najpodložnijih za kontaminaciju mikotoksinima. Aflatoksinima se, u poređenju s drugim mikotoksinima, pridaje najveća pažnja zbog njihove potencijalne kancerogenosti, značajnih i dugoročnih problema koje izazivaju kod ljudi i životinja. Vrste A. flavus i A. parasiticus mogu produkovati aflatoksine kod mnogih ekonomski značajnih kultura u poljima i skladištima. Zbog potencijalne kontaminacije zrna kukuruza aflatoksinima u ovom radu je ispitan toksigeni potencijal izolata upravo ove dve vrste poreklom iz Srbije i upoređene su različite metode detekcije ovih mikotoksina. Tokom eksperimenta upoređene su odgajivačke, serološke i analitičke metode detekcije mikotoksina, direktnom ekstrakcijom aflatoksina iz hranljive podloge. Istraživanja su bazirana na primeni odgajivačke metode za određivanje produkcije aflatoksina kod 20 izolata A. parasiticus (MRIZP Ap1-17) i A. flavus (MRIZP Af18-20) poreklom iz Srbije. Odgajivačke metode su bile zasnovane na formiranju žutog pigmenta u miceliji i hranljivoj podlozi, na pojavi fluorescencije na PDA (krompir dekstrozni agar) i podlozi koja sadrzi β-ciklodekstrin (CD-PDA), kao i na obrazovanju crvenog pigmenta u podlozi nakon dodavanja amonijum hidroksida. ELISA test je korišćen za proveru kvantitativnih i kvalitativnih sadržaja ukupnih aflatoksina B1, B2, G1 i G 2, dok je HPLC metodom utvrđena koncentracija pojedinačnih aflatoksina B1, B2, G1 i G2. Obrazovanje žutog pigmenta, fluorescencija i promena boje kolonije izolata u crvenu, kao dokaz toksigenosti izolata, potvrđena je u svim slučajevima i ELISA testom. Kod većine izolata ustanovljen je visok potencijal produkcije ukupnih aflatoksina. HPLC metodom potvrđena je i sposobnost sinteze aflatoksina G1 i G2 od strane izolata A. parasiticus. Cilj eksperimenta bio je da se ispita efikasnost upotrebe brzih testova za detekciju aflatoksina, kako bi se sprečila kontaminacija useva i izloženost ljudi i životinja afla-toksinima.
PB  - Matica srpska, Novi Sad
T2  - Zbornik Matice srpske za prirodne nauke
T1  - Comparison of methods for determination of the toxigenic potential of Aspergillus parasiticus Sp. and Aspergillus flavus L. isolated from maize
T1  - Komparacija metoda za utvrđivanje toksigenog potencijala Aspergillus parasiticus Speare i Aspergillus flavus Link izolovanih sa kukuruza
IS  - 133
SP  - 95
EP  - 104
DO  - 10.2298/ZMSPN1733095N
ER  - 
@article{
author = "Nikolić, Milica and Stanković, Slavica and Savić, Iva",
year = "2017",
url = "http://rik.mrizp.rs/handle/123456789/666",
abstract = "Maize is considered one of the most susceptible crops to mycotoxins worldwide. Compared to other mycotoxins, the greatest attention has been paid to aflatoxins, due to their potential carcinogenicity and due to significant and longstanding problems they can cause in humans and animals. A. flavus and A. parasiticus produce aflatoxins in many economically significant crops in both fields and storages. Because of the potential aflatoxin contamination of maize grain, the toxigenic potential of A. flavus and A. parasiticus isolates, originating from Serbia, was tested in the present study. Furthermore, various applied methods for detection of these mycotoxins were compared in the study. Cultural, serological and analytical methods for the detection of mycotoxins were compared in the course of the experiment by the direct extraction of aflatoxins from the nutrient medium. The cultural methods for the detection of aflatoxin production were applied to 20 isolates of A. flavus (MRIZP Af18-20) and A. parasiticus (MRIZP Ap1-17). These methods are based on the yellow pigment formation in mycelia and nutrition media, occurrence of fluorescence on PDA (potato dextrose agar), agar containing β-cyclodextrine (CD-PDA), as well as on the red pigment formation after adding ammonium hydroxide to the existing medium. The ELISA was used to check quantitative and qualitative analyses of total aflatoxins (B1, B2, G1, G2) while the HPLC method was applied to establish ability of isolates to synthesize aflatoxins B1, B2, G1, G2. The yellow pigment formation, fluorescence and colony color changes of isolates into red, as a proof of toxigenicity of isolates, were confirmed in all cases by ELISA. A high potential of total aflatoxin production was determined in the majority of observed isolates. The ability of A. parasiticus isolates to synthesize aflatoxins G1 and G2 was confirmed by the HPLC method. This was essential for a better understanding of the key role of the suitability of cultural methods for preliminary evaluation of a large number of isolates. Our goal was to employ rapid biochemical approaches to prevent aflatoxin contamination of crops, and to reduce human and animal exposure to foodborne mycotoxins., Kukuruz se širom sveta smatra jednim od useva najpodložnijih za kontaminaciju mikotoksinima. Aflatoksinima se, u poređenju s drugim mikotoksinima, pridaje najveća pažnja zbog njihove potencijalne kancerogenosti, značajnih i dugoročnih problema koje izazivaju kod ljudi i životinja. Vrste A. flavus i A. parasiticus mogu produkovati aflatoksine kod mnogih ekonomski značajnih kultura u poljima i skladištima. Zbog potencijalne kontaminacije zrna kukuruza aflatoksinima u ovom radu je ispitan toksigeni potencijal izolata upravo ove dve vrste poreklom iz Srbije i upoređene su različite metode detekcije ovih mikotoksina. Tokom eksperimenta upoređene su odgajivačke, serološke i analitičke metode detekcije mikotoksina, direktnom ekstrakcijom aflatoksina iz hranljive podloge. Istraživanja su bazirana na primeni odgajivačke metode za određivanje produkcije aflatoksina kod 20 izolata A. parasiticus (MRIZP Ap1-17) i A. flavus (MRIZP Af18-20) poreklom iz Srbije. Odgajivačke metode su bile zasnovane na formiranju žutog pigmenta u miceliji i hranljivoj podlozi, na pojavi fluorescencije na PDA (krompir dekstrozni agar) i podlozi koja sadrzi β-ciklodekstrin (CD-PDA), kao i na obrazovanju crvenog pigmenta u podlozi nakon dodavanja amonijum hidroksida. ELISA test je korišćen za proveru kvantitativnih i kvalitativnih sadržaja ukupnih aflatoksina B1, B2, G1 i G 2, dok je HPLC metodom utvrđena koncentracija pojedinačnih aflatoksina B1, B2, G1 i G2. Obrazovanje žutog pigmenta, fluorescencija i promena boje kolonije izolata u crvenu, kao dokaz toksigenosti izolata, potvrđena je u svim slučajevima i ELISA testom. Kod većine izolata ustanovljen je visok potencijal produkcije ukupnih aflatoksina. HPLC metodom potvrđena je i sposobnost sinteze aflatoksina G1 i G2 od strane izolata A. parasiticus. Cilj eksperimenta bio je da se ispita efikasnost upotrebe brzih testova za detekciju aflatoksina, kako bi se sprečila kontaminacija useva i izloženost ljudi i životinja afla-toksinima.",
publisher = "Matica srpska, Novi Sad",
journal = "Zbornik Matice srpske za prirodne nauke",
title = "Comparison of methods for determination of the toxigenic potential of Aspergillus parasiticus Sp. and Aspergillus flavus L. isolated from maize, Komparacija metoda za utvrđivanje toksigenog potencijala Aspergillus parasiticus Speare i Aspergillus flavus Link izolovanih sa kukuruza",
number = "133",
pages = "95-104",
doi = "10.2298/ZMSPN1733095N"
}
Nikolić, M., Stanković, S.,& Savić, I. (2017). Komparacija metoda za utvrđivanje toksigenog potencijala Aspergillus parasiticus Speare i Aspergillus flavus Link izolovanih sa kukuruza.
Zbornik Matice srpske za prirodne nauke
Matica srpska, Novi Sad.(133), 95-104.
https://doi.org/10.2298/ZMSPN1733095N
Nikolić M, Stanković S, Savić I. Komparacija metoda za utvrđivanje toksigenog potencijala Aspergillus parasiticus Speare i Aspergillus flavus Link izolovanih sa kukuruza. Zbornik Matice srpske za prirodne nauke. 2017;(133):95-104
Nikolić Milica, Stanković Slavica, Savić Iva, "Komparacija metoda za utvrđivanje toksigenog potencijala Aspergillus parasiticus Speare i Aspergillus flavus Link izolovanih sa kukuruza" Zbornik Matice srpske za prirodne nauke, no. 133 (2017):95-104,
https://doi.org/10.2298/ZMSPN1733095N .

Pathogenicity of Aspergillus spp. isolates originating from Serbia

Nikolić, Milica; Savić, Iva; Stanković, Slavica

(Institut za ratarstvo i povrtarstvo, Novi Sad, 2016)

TY  - JOUR
AU  - Nikolić, Milica
AU  - Savić, Iva
AU  - Stanković, Slavica
PY  - 2016
UR  - http://rik.mrizp.rs/handle/123456789/645
AB  - Species of the genus Aspergillus, section Flavi are facultative plant pathogens with the capability to contaminate primary agricultural products in the field, at harvest, in warehouses, and during processing. Infection is done through silk or injuries on maize ears. The objective of this study was to test pathogenicity of 25 A. parasiticus isolates on barley leaves. A parasiticus was isolated from maize kernels collected from 22 locations in Serbia during 2013-2015. Inoculation of the first barley leaf was done by the spore suspension of the fungus A. parasiticus. The spot sizes were statistically compared and this comparison showed that the isolate MRIZP 3930 was the least virulent. The isolate MRIZP 3819 was the most virulent seven days after inoculation and together with the isolate MRIZP 4265 was classified into the most aggressive group rated as 5. Pathogenicity was confirmed in all isolates.
AB  - Vrste roda Aspergillus sekcije Flavi su fakultativní biljni patogeni, čija se sposobnost ogleda u kontaminaciji primarnih poljoprivrednih proizvoda u polju, tokom berbe, u skladištu i tokom procesa prerade. Infekcija se obavlja preko svile ili oštećenja na klipu kukuruza. Cilj ovog rada bio je da se oceni patogenost 25 izolata A. parasiticus na listovima ječma. Kulture A. parasiticus su izolovane iz uzoraka zrna kukuruza koji su prikupljani sa 22 lokaliteta u Srbiji u periodu 2013-2015. godina. Inokulacija prvog lista ječma izvršena je pomoću suspenzije spora gljive A. parasiticus. Statističkim poređenjem dobijenih vrednosti veličina pega, pokazano je da je najmanje virulentan izolat MRIZP 3930. Izolat MRIZP 3819 se izvojio kao najvirulentniji posle 7 dana inokulacije, a zajedno s izolatom MRIZP 4265 je svrstan u najagresivniju grupu koja je kategorisana ocenom 5. Svi ispitivani izolati su potvrdili patogenost.
PB  - Institut za ratarstvo i povrtarstvo, Novi Sad
T2  - Ratarstvo i povrtarstvo
T1  - Pathogenicity of Aspergillus spp. isolates originating from Serbia
T1  - Patogenost izolata Aspergillus spp. poreklom iz Srbije
VL  - 53
IS  - 3
SP  - 101
EP  - 105
DO  - 10.5937/ratpov53-10795
ER  - 
@article{
author = "Nikolić, Milica and Savić, Iva and Stanković, Slavica",
year = "2016",
url = "http://rik.mrizp.rs/handle/123456789/645",
abstract = "Species of the genus Aspergillus, section Flavi are facultative plant pathogens with the capability to contaminate primary agricultural products in the field, at harvest, in warehouses, and during processing. Infection is done through silk or injuries on maize ears. The objective of this study was to test pathogenicity of 25 A. parasiticus isolates on barley leaves. A parasiticus was isolated from maize kernels collected from 22 locations in Serbia during 2013-2015. Inoculation of the first barley leaf was done by the spore suspension of the fungus A. parasiticus. The spot sizes were statistically compared and this comparison showed that the isolate MRIZP 3930 was the least virulent. The isolate MRIZP 3819 was the most virulent seven days after inoculation and together with the isolate MRIZP 4265 was classified into the most aggressive group rated as 5. Pathogenicity was confirmed in all isolates., Vrste roda Aspergillus sekcije Flavi su fakultativní biljni patogeni, čija se sposobnost ogleda u kontaminaciji primarnih poljoprivrednih proizvoda u polju, tokom berbe, u skladištu i tokom procesa prerade. Infekcija se obavlja preko svile ili oštećenja na klipu kukuruza. Cilj ovog rada bio je da se oceni patogenost 25 izolata A. parasiticus na listovima ječma. Kulture A. parasiticus su izolovane iz uzoraka zrna kukuruza koji su prikupljani sa 22 lokaliteta u Srbiji u periodu 2013-2015. godina. Inokulacija prvog lista ječma izvršena je pomoću suspenzije spora gljive A. parasiticus. Statističkim poređenjem dobijenih vrednosti veličina pega, pokazano je da je najmanje virulentan izolat MRIZP 3930. Izolat MRIZP 3819 se izvojio kao najvirulentniji posle 7 dana inokulacije, a zajedno s izolatom MRIZP 4265 je svrstan u najagresivniju grupu koja je kategorisana ocenom 5. Svi ispitivani izolati su potvrdili patogenost.",
publisher = "Institut za ratarstvo i povrtarstvo, Novi Sad",
journal = "Ratarstvo i povrtarstvo",
title = "Pathogenicity of Aspergillus spp. isolates originating from Serbia, Patogenost izolata Aspergillus spp. poreklom iz Srbije",
volume = "53",
number = "3",
pages = "101-105",
doi = "10.5937/ratpov53-10795"
}
Nikolić, M., Savić, I.,& Stanković, S. (2016). Patogenost izolata Aspergillus spp. poreklom iz Srbije.
Ratarstvo i povrtarstvo
Institut za ratarstvo i povrtarstvo, Novi Sad., 53(3), 101-105.
https://doi.org/10.5937/ratpov53-10795
Nikolić M, Savić I, Stanković S. Patogenost izolata Aspergillus spp. poreklom iz Srbije. Ratarstvo i povrtarstvo. 2016;53(3):101-105
Nikolić Milica, Savić Iva, Stanković Slavica, "Patogenost izolata Aspergillus spp. poreklom iz Srbije" Ratarstvo i povrtarstvo, 53, no. 3 (2016):101-105,
https://doi.org/10.5937/ratpov53-10795 .
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