Serial passage of the opportunistic pathogen Aspergillus flavus through an insect host yields decreased saprobic capacity

2005 ◽  
Vol 51 (2) ◽  
pp. 185-189 ◽  
Author(s):  
Lisa R Scully ◽  
Michael J Bidochka

To study the early stages of the effect of host restriction on pathogen evolution, we subjected the opportunistic fungus Aspergillus flavus to a serial propagation scheme, whereby insect-virulent conidia were selected for repeated passage through an insect host (Galleria mellonella larvae) for 5 generations. Of the 35 lineages promulgated through this scheme, there were no consistent changes in virulence, which was measured by percent mortality of the larvae. There were, however, increases in the number of conidia on the insect cadavers (9 of 35 lineages) and decreases in the number of days between death and the appearance of fungal growth on the cadavers (4 of 35 lineages). Notably, most of the lineages (28 of 35 lineages) demonstrated a statistically significant decrease in the diameter of the colonies subcultured onto artificial media, indicating a decreased ability to grow saprobically. Conversely, most of the A. flavus cultures successively grown on agar media (9 of 10 lineages) exhibited no change in colony diameter after 15 rounds of subculturing. Propagation of the opportunist A. flavus through the insect host G. mellonella resulted in a diminished capacity to grow on an alternate substrate, while maintaining or increasing its ability to use the host as a nutrient supply.Key words: Aspergillus flavus, insect pathogen, Galleria mellonella, serial propagation, emerging diseases.

Microbiology ◽  
2006 ◽  
Vol 152 (1) ◽  
pp. 223-232 ◽  
Author(s):  
Lisa R. Scully ◽  
Michael J. Bidochka

The evolution of host specialization in pathogens is a topic of considerable interest, particularly since it can represent a decisive step in the emergence of infectious diseases. Aspergillus flavus is an opportunistic fungus capable of infecting a wide variety of hosts, including plants, insects and mammals, although with low virulence. Here the derivation of an A. flavus strain that exhibits severe host restriction is reported. This strain exhibited a severe diminution or a complete lack of conidial production on a variety of standard agar media and on various plant species. However, it retained its ability to infect insects from various orders and to re-emerge from and adequately conidiate on the insect cadavers as a culmination of the pathogenic life cycle. This strain, demonstrating insect-dependent conidiation, was discovered to be a cysteine/methionine auxotroph due to an inability to reduce sulfate to sulfite. However, other A. flavus auxotrophs tested for plant and insect host range failed to show insect-dependent conidiation. An association between this specific auxotroph and a decreased host range is shown, emphasizing the role of nutrition in the host–pathogen relationship with respect to host restriction and evolution towards obligate pathogenesis.


Toxins ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 391
Author(s):  
Christopher Hernandez ◽  
Laura Cadenillas ◽  
Anwar El Maghubi ◽  
Isaura Caceres ◽  
Vanessa Durrieu ◽  
...  

Aflatoxin B1 (AFB1) is a potent carcinogenic mycotoxin that contaminates numerous crops pre- and post-harvest. To protect foods and feeds from such toxins without resorting to pesticides, the use of plant extracts has been increasingly studied. The most interesting candidate plants are those with strong antioxidative activity because oxidation reactions may interfere with AFB1 production. The present study investigates how an aqueous extract of Mimosa tenuiflora bark affects both the growth of Aspergillus flavus and AFB1 production. The results reveal a dose-dependent inhibition of toxin synthesis with no impact on fungal growth. AFB1 inhibition is related to a down-modulation of the cluster genes of the biosynthetic pathway and especially to the two internal regulators aflR and aflS. Its strong anti-oxidative activity also allows the aqueous extract to modulate the expression of genes involved in fungal oxidative-stress response, such as msnA, mtfA, atfA, or sod1. Finally, a bio-guided fractionation of the aqueous extract demonstrates that condensed tannins play a major role in the anti-aflatoxin activity of Mimosa tenuiflora bark.


Virulence ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 638-653
Author(s):  
Anne Marie Krachler ◽  
Natalie Sirisaengtaksin ◽  
Pauline Monteith ◽  
C. E. Timothy Paine ◽  
Christopher J. Coates ◽  
...  

Toxins ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 646 ◽  
Author(s):  
García-Díaz ◽  
Patiño ◽  
Vázquez ◽  
Gil-Serna

Aflatoxin (AF) contamination of maize is a major concern for food safety. The use of chemical fungicides is controversial, and it is necessary to develop new effective methods to control Aspergillus flavus growth and, therefore, to avoid the presence of AFs in grains. In this work, we tested in vitro the effect of six essential oils (EOs) extracted from aromatic plants. We selected those from Satureja montana and Origanum virens because they show high levels of antifungal and antitoxigenic activity at low concentrations against A. flavus. EOs are highly volatile compounds and we have developed a new niosome-based encapsulation method to extend their shelf life and activity. These new formulations have been successfully applied to reduce fungal growth and AF accumulation in maize grains in a small-scale test, as well as placing the maize into polypropylene woven bags to simulate common storage conditions. In this latter case, the antifungal properties lasted up to 75 days after the first application.


2015 ◽  
Vol 81 (18) ◽  
pp. 6129-6144 ◽  
Author(s):  
Abdulsamie Hanano ◽  
Ibrahem Almousally ◽  
Mouhnad Shaban ◽  
Elizabeth Blee

ABSTRACTCaleosins are a small family of calcium-binding proteins endowed with peroxygenase activity in plants. Caleosin-like genes are present in fungi; however, their functions have not been reported yet. In this work, we identify a plant caleosin-like protein inAspergillus flavusthat is highly expressed during the early stages of spore germination. A recombinant purified 32-kDa caleosin-like protein supported peroxygenase activities, including co-oxidation reactions and reduction of polyunsaturated fatty acid hydroperoxides. Deletion of the caleosin gene prevented fungal development. Alternatively, silencing of the gene led to the increased accumulation of endogenous polyunsaturated fatty acid hydroperoxides and antioxidant activities but to a reduction of fungal growth and conidium formation. Two key genes of the aflatoxin biosynthesis pathway,aflRandaflD, were downregulated in the strains in whichA. flavusPXG(AfPXG) was silenced, leading to reduced aflatoxin B1 productionin vitro. Application of caleosin/peroxygenase-derived oxylipins restored the wild-type phenotype in the strains in whichAfPXGwas silenced.PXG-deficientA. flavusstrains were severely compromised in their capacity to infect maize seeds and to produce aflatoxin. Our results uncover a new branch of the fungal oxylipin pathway and may lead to the development of novel targets for controlling fungal disease.


2021 ◽  
pp. 108201322110530
Author(s):  
Hanan H Abdel-Khalek ◽  
Ali AI Hammad ◽  
Reham MMA El-Kader ◽  
Khayria A Youssef ◽  
Dalia AM Abdou

The purpose of this study was to investigate the effects of certain essential oils (star anise, lemon leaves, marjoram, fennel, and lavender) on the fungal growth of Aspergillus flavus and Aspergillus parasiticus and their production of aflatoxin B1 (AFB1). The degree of suppression of the aflatoxigenic strains’ growth and their production of AFB1 is mainly affected by the kind and the concentration of the tested essential oils (EOs). Star anise essential oil had the lowest minimum inhibitory concentration (0.5 and 1.0 μL/mL) against A. flavus and A. parasiticus, respectively, so it was the best among the five different oils. The study of liquid chromatography with tandem mass spectrometry revealed that star anise EO resulted in a 98% reduction in AFB1 without a breakdown of AFB1 products after treatment thus the complete removal of AFB1 was done without any toxic residues. The combination showed a synergistic effect, the combinational treatment between γ-irradiation at a low dose (2 kGy) and star anise EO at concentrate 0.5 μL/g destroyed A. flavus and A. parasiticus inoculated (individually) in sorghum and peanut, respectively throughout the storage period (8 weeks).


2013 ◽  
Vol 2 (4) ◽  
pp. 68 ◽  
Author(s):  
Saifeldin Ahmed El-nagerabi ◽  
Abdulkadir E. Elshafie ◽  
Mohamed R. Elamin

<p>Aflatoxin and especially aflatoxin B<sub>1</sub> (AFB<sub>1</sub>) is a carcinogenic secondary metabolite synthesized by certain <em>Aspergillus </em>species. They contaminate natural and processed agricultural and animal products which render them unfit for consumption. The aim of this study was to evaluate the <em>in vitro</em> effects of <em>Balanites aegyptiaca</em> and <em>Tamarindus indica</em> fruit extracts on the growth and aflatoxin secretion of <em>Aspergillus flavus</em> (SQU21) and <em>A. parasiticus </em>(CBS921.7) strains. The two fruit extracts significantly (<em>P </em>&lt; 0.05) reduced aflatoxin and did not inhibit mycelial dry weights of the two <em>Aspergillus </em>strains. At different concentrations of balanites (2.5-10%), the inhibition of total aflatoxin was 49.9-84.8% for <em>A. flavus</em> (SQU21) and 32.1-84.4% for <em>A. parasiticus</em> (CBS921.7), whereas the inhibition of aflatoxin Bwas 38.2-81.4% and 32.8-80.6% for the two strains. Tamarind fruit extract (2.5-7.5%) caused 28.8-84.2% and 40.7-85.5% reductions in total aflatoxin and 37.1-83.5% and 33.9-85.9% in aflatoxin B for the two strains, respectively. None of these extracts inhibited the fungal growth or detoxified synthetic aflatoxin B<sub>1</sub>. We have concluded that these fruits contain various inhibitors to aflatoxin biosynthesis and secretion. Therefore, they can be used in combination as safe green biopreservatives to combat aflatoxin contamination of food.</p>


2013 ◽  
Vol 6 (1) ◽  
pp. 43-50 ◽  
Author(s):  
V. Aiko ◽  
A. Mehta

Cinnamon, cardamom, star anise and clove were studied for their effect on growth of Aspergillus flavus and aflatoxin B1 (AFB1) synthesis. The experiments were carried out in yeast extract sucrose culture broth as well as in rice supplemented with spices. AFB1 produced was analysed qualitatively and quantitatively using thin layer chromatography and high performance liquid chromatography, respectively. At a concentration of 10 mg/ml, cardamom and star anise did not exhibit any antifungal or anti-aflatoxigenic activity in culture broth, whereas cinnamon and clove inhibited A. flavus growth completely. The minimum inhibitory concentrations of cinnamon and clove were 4 and 2 mg/ml, respectively. Concentrations of cinnamon and clove below their minimum inhibitory concentrations showed enhanced fungal growth, while AFB1 synthesis was reduced. Clove inhibited the synthesis of AFB1 significantly up to 99% at concentrations ≥1.0 mg/ml. The spices also inhibited AFB1 synthesis in rice at 5 mg/g, although fungal growth was not inhibited. Clove and cinnamon inhibited AFB1 synthesis significantly up to 99 and 92%, respectively, and star anise and cardamom by 41 and 23%, respectively. The results of this study suggest the use of whole spices rather than their essential oils for controlling fungal and mycotoxin contamination in food grains.


Food Research ◽  
2019 ◽  
Vol 4 (1) ◽  
pp. 234-243
Author(s):  
M.M. Lisangan ◽  
R. Syarief ◽  
W.P. Rahayu ◽  
O.S. Dharmaputra

The objective of this study was to investigate the antifungal activity of kebar grass (Biophytum petersianum Klotzsch) extract (KGE) on the mycelial growth, conidiation and morphological structure of two types of aflatoxigenic fungus, which are Aspergillus flavus BCCF 0219 and A. flavus BIO 2236. They were isolated in the three types of model media, namely carbohydrate-enriched medium, fat-enriched medium and protein-enriched medium with five concentrations of KGE (12, 14, 16, 18, and 20 mg/mL) on each media. The best extract concentration of that inhibits the growth of A. flavus BCCF 0219 was found in the carbohydrate-enriched medium (95.7%), which was 12 mg/mL, whereas at A. flavus BIO 2236 was found in the fat-enriched medium (100%), which was 16 mg/mL. Based on SEM observation results, it was found that the mechanisms involved in fungal growth inhibitory by the KGE were by morphological alterations of the hyphal development, and the collapse of the entire hyphae. These findings indicated that KGE as a potential natural antifungal agent, particularly against aflatoxigenic fungi.


2002 ◽  
Vol 65 (12) ◽  
pp. 1984-1987 ◽  
Author(s):  
J. E. MELLON ◽  
P. J. COTTY

Soybean lines lacking lipoxygenase (LOX) activity were compared with soybean lines having LOX activity for the ability to support growth and aflatoxin B1 production by the fungal seed pathogen Aspergillus flavus. Whole seeds, broken seeds, and heat-treated (autoclaved) whole seeds were compared. Broken seeds, irrespective of LOX presence, supported excellent fungal growth and the highest aflatoxin levels. Autoclaved whole seeds, with or without LOX, produced good fungal growth and aflatoxin levels approaching those of broken seeds. Whole soybean seeds supported sparse fungal growth and relatively low aflatoxin levels. There was no significant difference in aflatoxin production between whole soybean seeds either with or without LOX, although there did seem to be differences among the cultivars tested. The heat treatment eliminated LOX activity (in LOX+ lines), yet aflatoxin levels did not change substantially from the broken seed treatment. Broken soybean seeds possessed LOX activity (in LOX+ lines) and yet yielded the highest aflatoxin levels. The presence of active LOX did not seem to play the determinant role in the susceptibility of soybean seeds to fungal pathogens. Seed coat integrity and seed viability seem to be more important characteristics in soybean seed resistance to aflatoxin contamination. Soybean seeds lacking LOX seem safe from the threat of increased seed pathogen susceptibility.


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