Nutritional impact of low-cost substrates on biphasic fermentation for conidia production of the fungal biopesticide Metarhizium anisopliae

Santos, P. de S.; Abati, Kauana; Mendoza, Natalie Verónika Rondinel; Mascarin, Gabriel Moura; Delalibera, Ítalo

doi:10.1016/j.biteb.2020.100619

Cited by 7 publications

(4 citation statements)

References 25 publications

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“…In this work, by employing a two-stage fermentation process for C. rosea, we were able to achieve a maximum yield of 1.1 × 10 9 conidia/gDM rice after 7 days of cultivation using parboiled rice as substrate. Various types of rice (white rice, parboiled rice, and broken rice) serve as both growth matrix and carbon and energy source in solid-state fermentation of many fungi used as biopesticides (Li et al, 2010;Mascarin et al, 2019;Santos P. S. et al, 2021). Nonetheless, further research on selecting agro-industrial wastes for C. rosea production by solid-state fermentation is needed to improve yields, shorten cultivation time, and reduce costs ensuring spore quality.…”

Section: Discussionmentioning

confidence: 99%

“…The solidstate fermentation method consists in the use of cereal grains as the main growth substrate, while the biphasic or two-stage fermentation process proposes a first step of growth by liquid culture which is subsequently used as inoculum source to attain the production of aerial conidia grown on the solid substrate. The solid-substrate fermentation presents bottlenecks, such as high costs of cereal-based substrates, intense demand for the labor, greater chance of contamination, automation deficiencies, long fermentation periods, and the lack of control of both nutritional and environmental conditions (Jackson, 1997;Mascarin et al, 2015;Santos P. S. et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

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Clonostachys rosea: Production by Submerged Culture and Bioactivity Against Sclerotinia sclerotiorum and Bemisia tabaci

Mascarin

Silva

et al. 2022

Front. Microbiol.

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Among the prospective biocontrol agents, the saprophytic filamentous fungus Clonostachys rosea is an excellent necrotrophic mycoparasite of numerous plant pathogenic fungi. However, its commercial development has been hampered by mass production difficulties during solid-state fermentation. Conversely, the submerged liquid fermentation shortens the cultivation time while increasing yields of fungal propagules. However, this method has been overlooked for C. rosea. In this work, we investigated the impact of liquid pre-culture inoculum on the spore production by the two-stage fermentation process using rice grains in comparison to the traditional solid-state fermentation. In parallel, we studied the submerged cultivation of C. rosea by manipulating carbon-to-nitrogen (C:N) ratio and nitrogen source, with the further optimization of spore production in a benchtop bioreactor. Additional bioassays included assessing the bioactivity of water-dispersible microgranules (that contained a submerged conidia) against the whitefly (Bemisia tabaci biotype B) and Sclerotinia sclerotiorum (causal agent of the white mold). Our results showed a maximum concentration of 1.1 × 109 conidia/g-dry-matter after 7 days of cultivation by two-stage fermentation process. The liquid fermentation yielded 1.4 × 109 submerged conidia/ml after 7 days using a medium with a 50:1 C:N ratio, and it also induced the production of microsclerotia (MS) up to 1.35 × 104/ml within 6 days with 10:1 C:N ratio; both media were supplemented with dextrose monohydrate and soybean meal. The fermentation batches carried out in a benchtop bioreactor with medium 50:1 C:N ratio and amended with soybean meal rendered a production peak on the fourth day, corresponding to 1.11 × 109 conidia/ml and 4.35 × 108 colony forming units (CFU)/ml. Following air-drying, the conidia production from air-dried microgranules of C. rosea biomass was estimated at 3.4 × 1010 conidia/g of formulated product upon re-hydration for 7 days. Both submerged conidia and MS of C. rosea inhibited 100% germination of S. sclerotiorum sclerotia by direct parasitism. The air-dried submerged conidia exhibited a suppressive activity on sclerotia (88% mycoparasitism) and early whitefly nymphs (76.2% mortality) that rendered LC50 values of 3.2 × 104 CFU/g soil and 1.5 × 107 CFU/ml, respectively. Therefore, the submerged liquid culture of C. rosea may offer a feasible and cost-effective method for its large-scale production, alleviating critical constraints to their commercial use while providing an additional tool for management of B. tabaci and S. sclerotiorum.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Clonostachys rosea: Production by Submerged Culture and Bioactivity Against Sclerotinia sclerotiorum and Bemisia tabaci

Mascarin

Silva

et al. 2022

Front. Microbiol.

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“…and 28 °C . In the same equipment, rice is then inoculated by sprinkling blastospore suspension on the particles while the drum rotates, up to a concentration of 10 8 blastospores/gds . The number of rotations and speed of rotation of the drum required to produce a homogeneous distribution of blastospore can be obtained following methodologies available in literature. , The production of blastospore suspension is described in section 3.5.…”

Section: Alternative Production Processmentioning

confidence: 99%

“…In contrast to aerial conidia, blastospores generated by liquid culture require a shorter incubation time and germinate faster. The blastopore production technique needs less labor and is more adequate for automation, increasing conidia production on solid substrate . However, the cost of liquid culture medium should be considered, so a low-cost carbon source, corn syrup, with an estimated value of US$0.30/L, is employed instead of the traditional ADAMEK culture medium, which costs US$2.10/L …”

Section: Alternative Production Processmentioning

confidence: 99%

Assessment of the Techno-Economic Feasibility of Semicontinuous Production of Metarhizium anisopliae Spores

Ferreira,

Mendes,

Thoméo

2023

Ind. Eng. Chem. Res.

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The significant increase and indiscriminate use of chemical pesticides have led to the development of several environmental problems and adverse effects on human health. Alternatively, bioinsecticides have been successfully used for a variety of crops to control several insect infestations with efficiency and competitive costs. Metarhizium anisopliae have widely been used in sugar cane crops in Brazil, and their use is the most successful enthomopathogen program worldwide. However, the production of this propagule is semiartisanal and presents several technical bottlenecks. In order to intensify this production, all the steps of the process have been revised and more efficient techniques and procedures have been adopted to avoid exogenous contamination and increase the productivity. The most relevant modification in this semicontinuous process is the cultivation of rice in packed-bed solid-state cultivation bioreactors and the extraction of spores in rotating drums, with retention of the spores in cyclones. The economic analysis showed that this process is competitive when compared with the traditional process of spore production and with similar biotechnological processes.

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Performance of Metarhizium rileyi Nm017: nutritional supplementation to improve production and quality conidia

Mejía,

Rocha,

Sanabria

et al. 2024

3 Biotech

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This study aimed to analyze the effect of nutritional supplements on improving conidia production of Metarhizium rileyi Nm017 at laboratory scale (yields of conidia/substrate and biomass/substrate, and substrate consumption). Also, the influence on quality parameters were evaluated (germination at 36 and 48 h, enzymatic activity, and insecticidal activity on Helicoverpa zea). Six treatments (T1–T6) were assessed and all of them reached maximum conidia concentration after 7 days fermentation, a feasible production timetable. Yields from treatment T6 (yeast extract + V8 juice) were 1.5–threefold higher than the other treatments. Conidia from T6 reached germinations of 56% and 12% at 36 and 48 h, respectively, higher than T1 (without supplements), which had the lowest values found. M. rileyi conidia obtained from treatment T6 had the highest enzymatic activity (0.45 U chitinase g−1, 0.28 U lipase g−1, and 1.29 U protease g−1). However, treatments with the highest conidia yields and enzymatic activity were not positively correlated to the efficacy against H. zea. When M. rileyi was produced on T5 (yeast hydrolysate + V8 juice), conidia were 35% more virulent than treatment T6. The findings evidenced the noticeable impact of nutritional substrate amended for conidia production and quality. This work showed the relevance of insecticidal activity assessment as a selection criterion in the mass production development of a biocontrol agent.

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Nutritional impact of low-cost substrates on biphasic fermentation for conidia production of the fungal biopesticide Metarhizium anisopliae

Cited by 7 publications

References 25 publications

Clonostachys rosea: Production by Submerged Culture and Bioactivity Against Sclerotinia sclerotiorum and Bemisia tabaci

Clonostachys rosea: Production by Submerged Culture and Bioactivity Against Sclerotinia sclerotiorum and Bemisia tabaci

Assessment of the Techno-Economic Feasibility of Semicontinuous Production of Metarhizium anisopliae Spores

Performance of Metarhizium rileyi Nm017: nutritional supplementation to improve production and quality conidia

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