scholarly journals The Development of a Phytopathogenic Fungi Control Trial: Aspergillus flavus and Aspergillus niger Infection in Jojoba Tissue Culture as a Model

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Nawal Abd El-Baky ◽  
Raoufa Ahmed Abdel Rahman ◽  
Mona Mohammed Sharaf ◽  
Amro Abd Al Fattah Amara

After introducing the idea of using concentrations equal to or less than the minimum inhibition concentration (MIC) of some active chemical compounds for evacuating microbial cells, different types of microbes were evacuated. The original protocol was given the name sponge-like protocol and then was reduced and modified from a microorganism to another to prepare microbial ghosts for various applications such as immunological applications, drug delivery, and isolation of DNA and protein. Fungal pathogens that infect plants critically affect cost effectiveness, quality, and quantity of their production. They kill plant cells and/or cause plant stress. Plant fungal infections can originate from many sources such as infected soil, seeds, or crop debris causing diseases and quality losses around the world with billions of US dollars annually as costs of the associated productivity loss. This study focused on the application of the sponge-like protocol in protecting in vitro tissue cultures of plants against fungal pathogens. This can be useful for research purposes or may be developed to be introduced in field applications. Aspergillus flavus and Aspergillus niger infection in tissue culture of jojoba (Simmondsia chinensis (Link) Schn.) was used as a model to establish the employment of this protocol to control plant fungal diseases. The best conditions for A. flavus and A. niger ghosts production previously mapped by randomization experimental design (reduced Plackett–Burman experimental design) were used to prepare fungal ghosts. SDS, NaOH, NaHCO3, and H2O2 were used in their MIC (+1 level) or minimum growth concentration (MGC, −1 level) according to the determined optimal experimental design. The release of both of DNA and protein from the fungal cells was evaluated spectrophotometrically at 260nm and 280nm, respectively, as an indicator for cell loss of their cytoplasm. Fungal ghost cells were also examined by transmission electron microscopy. After confirming the preparation of high-quality fungal ghost cells, the same conditions were mimicked to control plant fungal infection. Jojoba grown in tissue culture was sprayed with fungal cells (about 103 CFU) as a control experiment or fungal cells followed by treatment with solution (a) represents the fungal ghost cells formation calculated critical concentration (FGCCC) of SDS, NaOH, and NaHCO3 and then treatment with solution (b) represents H2O2 FGCCC. The plant was examined on day 0 (plant grown before any infection or infection followed by treatment), day 5 (plant at day 5 after infection or infection followed by treatment), and day 10 (plant at day 10 after infection or infection followed by treatment). We observed fungal growth in case of control experiments at days 5 and 10 on the tissue culture medium, as well as plant, and the absence of any fungal growth in case of plant treated with FGCCC even after day 10. We recommend using this FGCCC in the form of chemical spraying formulation to treat the plants aiming to control different plant fungal infections in in vitro tissue culture systems or applied in field.

2021 ◽  
pp. 55-66
Author(s):  
Roxanne Joy Colendres ◽  
Carl Leonard Pradera

Fungal infections caused by Trichophyton mentagrophytes, Aspergillus fumigatus , and Malassezia pachydermatis are among the major contributors to multisystemic health problems such as dermatitis, otitis, and respiratory disorders among humans and animals. This study was conducted to determine the in vitro antifungal activity of Terminalia catappa leaf crude aqueous and ethanolic extracts against these fungal pathogens by measuring the zone of inhibition (ZI) using the agar well diffusion technique. Qualitative phytochemical screening tests were also performed to determine bioactive compounds present in the plant extract. Results show that the plant’s crude aqueous (CAE) and ethanolic extracts (CEE) were found to be effective against all test fungi. M. pachydermatis showed susceptibility towards CAE and CEE from T1 (100%), T2 (75%), T3 (50%) and T4 (25%), with the highest mean ZI of 18.33mm and 13.33, respectively. On the other hand, T. mentagrophytes was inhibited by CAE and CEE at T1 (100%), T2 (75%) and T3 (50%) with the highest mean ZI of 9.67mm and 10.33mm, respectively. At the same time, it was observed that A. fumigatus had reactive sensitivity towards CAE and CEE at T1 (100%) and T2 (75%), with the highest mean ZI of 9.33mm and 10.33mm, respectively. Moreover, phytochemical tests showed that the plant’s leaf crude extracts contain alkaloids, saponins, and tannins, which could potentially inhibit fungal growth.


Plants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 114
Author(s):  
Armina Morkeliūnė ◽  
Neringa Rasiukevičiūtė ◽  
Lina Šernaitė ◽  
Alma Valiuškaitė

The Colletotrichum spp. is a significant strawberry pathogen causing yield losses of up to 50%. The most common method to control plant diseases is through the use of chemical fungicides. The findings of plants antimicrobial activities, low toxicity, and biodegradability of essential oils (EO), make them suitable for biological protection against fungal pathogens. The aim is to evaluate the inhibition of Colletotrichum acutatum by thyme, sage, and peppermint EO in vitro on detached strawberry leaves and determine EO chemical composition. Our results revealed that the dominant compound of thyme was thymol 41.35%, peppermint: menthone 44.56%, sage: α,β-thujone 34.45%, and camphor: 20.46%. Thyme EO inhibited C. acutatum completely above 200 μL L−1 concentration in vitro. Peppermint and sage EO reduced mycelial growth of C. acutatum. In addition, in vitro, results are promising for biological control. The detached strawberry leaves experiments showed that disease reduction 4 days after inoculation was 15.8% at 1000 μL L−1 of peppermint EO and 5.3% at 800 μL L−1 of thyme compared with control. Our findings could potentially help to manage C. acutatum; however, the detached strawberry leaves assay showed that EO efficacy was relatively low on tested concentrations and should be increased.


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>


2021 ◽  
Vol 30 (3) ◽  
pp. 127-134
Author(s):  
Shaimaa A.S. Selem ◽  
Neveen A. Hassan ◽  
Mohamed Z. Abd El-Rahman ◽  
Doaa M. Abd El-Kareem

Background: In intensive care units, invasive fungal infections have become more common, particularly among immunocompromised patients. Early identification and starting the treatment of those patients with antifungal therapy is critical for preventing unnecessary use of toxic antifungal agents. Objective: The aim of this research is to determine which common fungi cause invasive fungal infection in immunocompromised patients, as well as their antifungal susceptibility patterns in vitro, in Assiut University Hospitals. Methodology: This was a hospital based descriptive study conducted on 120 patients with clinical suspicion of having fungal infections admitted at different Intensive Care Units (ICUs) at Assiut University Hospitals. Direct microscopic examination and inoculation on Sabouraud Dextrose Agar (SDA) were performed on the collected specimens. Isolated yeasts were classified using phenotypic methods such as chromogenic media (Brilliance Candida agar), germ tube examination, and the Vitek 2 system for certain isolates, while the identification of mould isolates was primarily based on macroscopic and microscopic characteristics. Moulds were tested in vitro for antifungal susceptibility using the disc diffusion, and yeast were tested using Vitek 2 device cards. Results: In this study, 100 out of 120 (83.3%) of the samples were positive for fungal infection. Candida and Aspergillus species were the most commonly isolated fungal pathogens. The isolates had the highest sensitivity to Amphotericin B (95 %), followed by Micafungin (94 %) in an in vitro sensitivity survey. Conclusion: Invasive fungal infections are a leading cause of morbidity and mortality in immunocompromised patients, with Candida albicans being the most frequently isolated yeast from various clinical specimens; however, the rise in resistance, especially to azoles, is a major concern.


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.


Antibiotics ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 188 ◽  
Author(s):  
Alberto Antonelli ◽  
Luca Giovannini ◽  
Ilaria Baccani ◽  
Valentina Giuliani ◽  
Riccardo Pace ◽  
...  

The recent increase in infections mediated by drug-resistant bacterial and fungal pathogens underlines the urgent need for novel antimicrobial compounds. In this study, the antimicrobial activity (inhibitory and cidal) of HybenX®, a novel dessicating agent, in comparison with commonly used sodium hypochlorite and chlorhexidine, against a collection of bacterial and yeast strains representative of the most common human pathogenic species was evaluated. The minimal inhibitory, bactericidal, and fungicidal concentrations (MIC, MBC, and MFC, respectively) of the three different antimicrobial agents were evaluated by broth microdilution assays, followed by subculturing of suitable dilutions. HybenX® was active against 26 reference strains representative of staphylococci, enterococci, Enterobacterales, Gram-negative nonfermenters, and yeasts, although at higher concentrations than sodium hypochlorite and chlorhexidine. HybenX® MICs were 0.39% for bacteria (with MBCs ranging between 0.39% and 0.78%), and 0.1–0.78% for yeasts (with MFCs ranging between 0.78% and 1.6%). HybenX® exhibited potent inhibitory and cidal activity at low concentrations against several bacterial and yeast pathogens. These findings suggest that HybenX® could be of interest for the treatment of parodontal and endodontic infections and also for bacterial and fungal infections of other mucous membranes and skin as an alternative to sodium hypochlorite and chlorhexidine.


2019 ◽  
Vol 82 (10) ◽  
pp. 1775-1782 ◽  
Author(s):  
XUEMEI LUO ◽  
HONGXIA LI ◽  
DAN JIANG ◽  
JIEQIN MENG ◽  
FAN ZHANG ◽  
...  

ABSTRACT Coix (Coix lacryma-jobi) seeds are susceptible to fungal infections, making their surface fungi complex and diverse. Some fungi can produce mycotoxins under suitable conditions, and fungal growth is closely related to the production of mycotoxins. In this study, the surface fungi of coix seed were identified by Illumina HiSeq high-throughput sequencing. Simultaneously, the fungi cultured by the plate method were identified by microscopy and DNA barcoding; finally, the species of fungi were identified accurately and reliably by combining three methods. The aqueous extract of coix seed was cocultured with Aspergillus flavus spores, and the relationship between the aqueous extract and the growth of A. flavus was studied with the dry weight of mycelium as an indicator. The results showed that there were 89 genera and 96 species of fungi on coix seed, which were mainly distributed in Ascomycota (81.48%) and Basidiomycota (4.08%), and Xeromyces (8.50%), Gibberella (7.25%), and Aspergillus (4.74%) were the predominant genera. Four fungi were isolated from coix seed by plate culture and identified as Aspergillus fumigatus, A. flavus, Aspergillus oryzae, and Rhizopus oryzae by microscopy and DNA barcoding. The aqueous extract of coix seed at low concentrations has a promoting effect on the growth of A. flavus. When the concentration is 3.125%, the promotion effect is the most pronounced, and the promotion rate is 29.17%. These results reveal the diversity of fungi on the coix seed, which can provide a reference for the prevention and control of harmful fungi on coix seed.


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