Detection of Aspergillus flavus in Wheat Grains Using Anti-mannoprotein (MP1) and Spore Protein Polyclonal Antibodies

2022 ◽  
Author(s):  
Ranjana Kumari ◽  
Ananta Kumar Ghosh
Keyword(s):  
Aspergillus Flavus ◽  
Polyclonal Antibodies ◽  
Wheat Grains

scholarly journals Detection of Aspergillus Flavus in Wheat Grains Using Anti-Mannoprotein (MP1) and Spore Proteins Polyclonal Antibodies

2021 ◽  
Author(s):  
Ranjana Kumari ◽  
Ananta Ghosh
Keyword(s):  
Aspergillus Flavus ◽  
Polyclonal Antibodies ◽  
Protein A ◽  
Fungal Species ◽  
Cross Reactivity ◽  
Soluble Form ◽  
Minimum Concentration ◽  
Wheat Grains ◽  
Stored Wheat ◽  
Spore Proteins

Abstract Cell wall mannoprotein (MP1) gene of an aflatoxigenic strain of Aspergillus flavus, isolated from stored wheat grains, was cloned and sequenced. MP1 protein was expressed in E. coli in soluble form and purified. Polyclonal antibodies were raised against recombinant MP1 protein and inactivated spores of this fungus in rabbit, and purified by ammonium sulphate precipitation, Protein A sepharose and antigen affinity chromatography. The minimum concentration of purified mycelial or spore proteins that could be detected by ELISA was determined as 100 ng using 2 µg of these antibodies. The anti-MP1 antibody was found more sensitive than anti-spore protein antibody. Western blot and immunofluorescence analysis showed reactivity of these antibodies to various proteins (30 kDa to 200 kDa) distributed throughout the surface of mycelia and spore of A. flavus. Cross reactivity of these antibodies was detected with fungi belonging to different Aspergillus, Rhizopus and Alternaria species out of fourteen different fungal species tested. In fungal contaminated wheat grains these antibodies could detect presence of as low as 1 µg mycelia or 103 spores per gram of wheat grains using ELISA. The results suggest that the developed antibodies could be successfully applied for the detection of predominant fungal infestation in stored wheat grains.

scholarly journals Detection the Ability of Aspergillus flavus Isolated from Wheat Grains for Aflatoxin B1 Production using RT- PCR

2015 ◽  
Vol 9 (2) ◽  
pp. 102-106
Author(s):  
Eman Noaman Ismail
Keyword(s):  
Molecular Weight ◽  
Triticum Aestivum ◽  
Aspergillus Flavus ◽  
Aflatoxin B1 ◽  
Reverse Transcription ◽  
Regulatory Gene ◽  
Rapid Assessment ◽  
Rt Pcr ◽  
Cdna Synthesis ◽  
Wheat Grains

Aspergillus flavus is aflatoxinogenic and potential aflatoxins producers in agriculturalcommodities. The present study was conducted determine the ability of eleven strains of A. flavusisolated from Iraqi wheat grains Triticum aestivum. The isolates have been detected by molecularmethods using Reverse Transcriptase RTPCR. In this study, RNA was extracted from A. flavus,cDNA synthesis and rapid assessment of eleven isolates of A. flavus was accomplished usingprimer pair for the aflatoxin regulatory gene aflR Reverse transcription-Polymerase chainreaction (RT–PCR).Positive amplification was achieved for all the isolates with a molecularweight 798 to aflR1 and 400bp to aflR2. Also the result of the amplification showed there are nodifferences with the two molecular weight between the 11 isolated strains of A. flavus in theiraflatoxin B1 production, but the first strain differed in their banding florescence as comparedwith others strains this reflect the genetic differences in aflatoxin B1 production between them.

scholarly journals Molecular Identification and Characterisation of Aspergillus Flavus Isolates Originating from Serbian Wheat Grains

2020 ◽  
Vol 49 (4) ◽  
pp. 382-389
Author(s):  
J. Krulj ◽  
N. Ćurčıć ◽  
A. Bočarov Stančıć ◽  
J. Kojıć ◽  
L. Pezo ◽  
...  
Keyword(s):  
Aspergillus Flavus ◽  
Molecular Genetic ◽  
Pcr Amplification ◽  
Its Region ◽  
Holistic Approach ◽  
Accurate Identification ◽  
Essential Point ◽  
Climate Conditions ◽  
Wheat Grains ◽  
Pcr Rflp

During previous years, regarding the shifts in climate conditions in temperate region, such as occurrence of high temperatures and prolonged drought, increased occurrence frequencies of Aspergillus flavus and aflatoxins in cereal grains were recorded. A reliable and accurate identification of the fungi is of great importance for evaluating the microbiological risks of contamination. The essential point of the present investigation was molecular characterisation and identification of A. flavus isolates originating from common wheat and spelt grains collected after harvest during the period of three years (2015–2017) in Northern Serbia. A holistic approach that included PCR amplification of two DNA genomic regions and PCR-RFLP assay followed by fragment length analysis, provided complete and comprehensive characterisation of A. flavus isolated from wheat grains. The presented results indicate that there was no difference among the tested Aspergillus isolates on the molecular–genetic level. All 38 strains were identified as A. flavus by sequencing of combined ITS region and β-tubulin gene fragments (acc. no.: MH582473 to MH582510). PCR-RFLP method in combination with a Lab-on-a-chip (LoaC) electrophoresis can be successfully used to rapidly identify A. flavus isolates.

scholarly journals Population dynamics of mycoflora and incidence of black point disease in wheat grains

1970 ◽  
Vol 35 (1) ◽  
pp. 1-10 ◽  
Author(s):  
PK Malaker ◽  
IH Mian
Keyword(s):  
Population Dynamics ◽  
Aspergillus Flavus ◽  
Exposure Period ◽  
Grain Development ◽  
Point Index ◽  
Fusarium Spp ◽  
Black Point ◽  
Wheat Grains ◽  
Definite Pattern ◽  
Developing Grains

An attempt was made to monitor the prevalence of different fungi associated with floret lemma and developing grains of wheat and to assess the subsequent incidence of black point disease under different exposure periods of the spikes to airborne inocula of the causal fungi. Altogether 16 fungi representing 11 genera were detected from lemma and developing grains. The predominant fungi, in order of prevalence, were A. alternata, B. sorokiniana, C. cladosporioides, C. lunata, Fusarium spp. and E. purpurascens. Other fungi occurring less commonly were A. triticina, C. pallescens, Nigrospora sp., Phoma sp., Chaetomium sp., B. tetramera, B. oryzae, Aspergillus flavus, A. niger and Doratomyces sp. The incidence of all the fungi except Fusarium spp. was higher in lemma than in grains. In lemma, A. alternata occurred with the highest frequency, while the incidence of B. sorokiniana was found highest in grains. The population of A. alternata, B. sorokiniana, C. lunata, E. purpurascens and Fusarium spp. increased with the age of lemma and developing grains whereas the incidence of C. cladosporioides increased at early stages but declined at later stages of grain development. Other fungi did not follow any definite pattern in their incidence during grain development. The occurrence of black pointed grains per spike, percent black pointed grains and black point index were found increasing with the increase in exposure period of the spikes to airborne inocula of the causal fungi. Keywords: Mycoflora; black point; wheat. DOI: 10.3329/bjar.v35i1.5861Bangladesh J. Agril. Res. 35(1) : 1-10, March 2010

scholarly journals Anti-Toxigenic Effect of Lactic Acid Bacteria Against Aspergillus spp Isolated from Wheat Grains

2020 ◽  
Vol 14 (1) ◽  
pp. 252-259
Author(s):  
Mohamed T. Fouad ◽  
Tarek A. El-Desouky
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Aspergillus Flavus ◽  
Dairy Products ◽  
Aspergillus Parasiticus ◽  
Lactobacillus Rhamnosus ◽  
Storage Conditions ◽  
Wheat Grain ◽  
Wheat Grains ◽  
Lactobacillus Gasseri

Introduction: Many fungi infect the wheat grains. Under field and or storage conditions from temperature and humidity, some fungi can produce aflatoxins (AFs), which may cause acute or chronic diseases. Therefore, there is a necessary and urgent need to find an effective and safe way to reduce or remove AFs. Objective: The objective of this study was the evaluation of Lactobacillus rhamnosus, Lactobacillus gasseri, and Lactobacillus plantarum for their ability to reduce and or remove AFs produced by Aspergillus flavus and Aspergillus parasiticus, which were isolated from wheat grains, as well as control of AFs produced on affected wheat grain by A.parasiticus spores only. Methods: LAB, isolated from some local dairy products, were cultured in MRS for the evaluation of their ability to remove AFs, produced by A. flavus and A. parasiticus on (YES) media, in addition to the treatment of wheat grains by LAB cells to prevent AFs produced by A. parasiticus. Results: The L. rhamnosus strain gave the highest reduction rates of AFs produced by A. parasiticus that were 62.6, 44.4, 43.3, and 52.2% for AFG1, AFB1, AFG2, and AFB2, respectively. While in the case of A. flavus, the reduction was 50.4, 42.7, 40.6, and 36.8% in the same order of toxins. When applied, these strains with wheat grains were affected by A. parasiticus, the inhibition rates of AFs were ranged between 61.4 and 75.8% with L. rhamnosus strain and 43.7 to 52.1% with L. gasseri, while L. plantarum strain ranged from 55.5 to 66.9%. Conclusion: According to this study, L. rhamnosus is considered one of the best strains in this field. Therefore, the present study suggests applied use of LAB as a treatment to prevent AFs production in wheat grains.

Immunocytochemical methodology for the localization of neuronal antigens at the ultrastructural level

1990 ◽  
Vol 48 (3) ◽  
pp. 876-877
Author(s):  
Jorge Pecci Saavedra ◽  
Mark Connaughton ◽  
Juan José López ◽  
Alicia Brusco
Keyword(s):  
Spinal Cord ◽  
Substantia Nigra ◽  
Polyclonal Antibodies ◽  
Ultrastructural Level ◽  
Point Of View ◽  
Nerve Tissue ◽  
Tissue Fixation ◽  
Optimal Conditions ◽  
Interpeduncular Nucleus ◽  
Technical Point

The use of antibodies as labels for the localization of specific molecules in the nervous systan has been extensively applied in recent years. Both monoand polyclonal antibodies or antisera have been employed. The knowledge of the organization of neuronal connectivities, gliovascular relationships, glioneuronal relationships and other features of nerve tissue has greatly increased.A number of areas of the nervous systan have been analyzed in our laboratory, including the nuclei of the raphe system, the reticular formation, interpeduncular nucleus, substantia nigra, caudate nucleus, putamen, pallidum, spinal cord, pineal gland and others.From a technical point of view, a number of variables needed to be taken into account in order to obtain reliable and reproducible results. The design of the optimal conditions of tissue fixation, embedding, sectioning, dilution of antibodies, and adaptation of Sternberger PAP technique were sane of the parameters taken into account to optimize the results. It is critical that each step of the technique be defined for each particular case.

Systemic aspergillosis caused by an aflatoxin-producing strain of Aspergillus flavus

1998 ◽  
Vol 36 (2) ◽  
pp. 107-112 ◽  
Author(s):  
T. MORI ◽  
M. MATSUMURA ◽  
K. YAMADA ◽  
S. IRIE ◽  
K. OSHIMI ◽  
...  
Keyword(s):  
Aspergillus Flavus

Transport of zinc and manganese to developing wheat grains

1995 ◽  
Vol 95 (3) ◽  
pp. 449-455 ◽  
Author(s):  
J. N. Pearson ◽  
Z. Rengel ◽  
C. F Jenner ◽  
R. D. Graham
Keyword(s):  
Wheat Grains
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