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Screening of Microorganisms Isolated from Stingless Bees' Larval Food in the Biocontrol of Meloidogyne incognita

Guilherme Nunes Moreira Costa

Guilherme Nunes Moreira Costa

Laboratory of Genetics, Institute of Biotechnology, Federal University of UberlândiaUberlândia, Brazil
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Nunes Moreira Costa, Guilherme
, 
Ana Carolina Costa Santos

Ana Carolina Costa Santos

Laboratory of Genetics, Institute of Biotechnology, Federal University of UberlândiaUberlândia, Brazil
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Santos, Ana Carolina Costa
, 
Tamires dos Santos Paschoal

Tamires dos Santos Paschoal

Laboratory of Genetics, Institute of Biotechnology, Federal University of UberlândiaUberlândia, Brazil
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dos Santos Paschoal, Tamires
, 
Anna Paula Martins Garcia

Anna Paula Martins Garcia

Laboratory of Genetics, Institute of Biotechnology, Federal University of UberlândiaUberlândia, Brazil
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Garcia, Anna Paula Martins
, 
Anderson Rodrigues dos Santos

Anderson Rodrigues dos Santos

Faculty of Computer Science, Federal University of UberlândiaUberlândia, Brazil
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dos Santos, Anderson Rodrigues
 and 
Carlos Ueira-Vieira

Carlos Ueira-Vieira

Laboratory of Genetics, Institute of Biotechnology, Federal University of UberlândiaUberlândia, Brazil
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Ueira-Vieira, Carlos
  
Jun 21, 2025
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Article Category: Research Paper
Published Online: Jun 21, 2025
Received: Feb 05, 2025
DOI: https://doi.org/10.2478/jofnem-2025-0028
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© 2025 Guilherme Nunes Moreira Costa et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

Nematicidal activity of microorganisms isolated from stingless bee larval food against M. incognita inoculated in pots with soybean. The negative control used water and the positive control used the commercial biocontrol product Nemat (recommended dosage). Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1) (Dotmatics, Boston, MA). *p < 0.05; **p < 0.01.
Nematicidal activity of microorganisms isolated from stingless bee larval food against M. incognita inoculated in pots with soybean. The negative control used water and the positive control used the commercial biocontrol product Nemat (recommended dosage). Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1) (Dotmatics, Boston, MA). *p < 0.05; **p < 0.01.

Figure 2:

The effect of isolate Mq-MCK-07 on M. incognita inoculated in pots with soybean. (A) Number of nematodes per root gram 60 days after inoculation. (B) Number of nematodes in the soybean roots 60 days after inoculation. Untreated pots with no nematode inoculation were set to check possible contaminations among treatments. The negative control used water and the positive control used the commercial biocontrol product Nemat (recommended dosage). Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1). *p < 0.05; **p < 0.01.
The effect of isolate Mq-MCK-07 on M. incognita inoculated in pots with soybean. (A) Number of nematodes per root gram 60 days after inoculation. (B) Number of nematodes in the soybean roots 60 days after inoculation. Untreated pots with no nematode inoculation were set to check possible contaminations among treatments. The negative control used water and the positive control used the commercial biocontrol product Nemat (recommended dosage). Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1). *p < 0.05; **p < 0.01.

Figure 3:

The Mq-MCK-07 bacteria inhibited M. incognita egg-hatching in vitro. Egg hatching rates were evaluated with 50 M. incognita eggs incubated at 28 °C for 72 h with different treatments. The negative control used distilled water. Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1). **p < 0.01; ***p < 0.001.
The Mq-MCK-07 bacteria inhibited M. incognita egg-hatching in vitro. Egg hatching rates were evaluated with 50 M. incognita eggs incubated at 28 °C for 72 h with different treatments. The negative control used distilled water. Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1). **p < 0.01; ***p < 0.001.

Figure 4:

The isolate Mq-MCK-07 showed a nematicidal effect on J2 of M. incognita in the in vitro test. Mortality rates were evaluated with 100 J2 of M. incognita incubated at 28 °C for 24 h with different treatments. The negative control was distilled water. Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1). ***p < 0.001.
The isolate Mq-MCK-07 showed a nematicidal effect on J2 of M. incognita in the in vitro test. Mortality rates were evaluated with 100 J2 of M. incognita incubated at 28 °C for 24 h with different treatments. The negative control was distilled water. Data analysis was performed using One-Way analysis of variance (ANOVA) and Tukey multiple comparison tests in the Graphpad Prism software (version 10.1.1). ***p < 0.001.

Figure 5:

Graphical circular map of Enterococcus faecalis strain mandacaium chromosome (2.61 Mb). From outer circle to the center: all CDS on forward and reverse strands, all CDS on forward strand, all CDS on reverse strand, all rRNAs on forward and reverse strands. Green: > 95% of identity with reference genome; Yellow: 70% to 95% of identity with reference genome; Red: < 70% of identity with reference genome; Black: rRNA sequences. Inner circle shows GC content (Blue: Below average; Orange: above average). The map was generated using DNAPlotter from Artemis (Carver et al., 2012).
Graphical circular map of Enterococcus faecalis strain mandacaium chromosome (2.61 Mb). From outer circle to the center: all CDS on forward and reverse strands, all CDS on forward strand, all CDS on reverse strand, all rRNAs on forward and reverse strands. Green: > 95% of identity with reference genome; Yellow: 70% to 95% of identity with reference genome; Red: < 70% of identity with reference genome; Black: rRNA sequences. Inner circle shows GC content (Blue: Below average; Orange: above average). The map was generated using DNAPlotter from Artemis (Carver et al., 2012).

Figure 6:

Graphical circular map of Enterococcus faecalis strain mandacaium plasmid 1 (20,406 bp). From outer circle to the center: all CDS on forward and reverse strands; all CDS on forward strand; all CDS on reverse strand. Green: > 95% of identity with reference genome; Yellow: 70% to 95% of identity with reference genome; Red: < 70% of identity with reference genome. Inner circle shows GC content (Blue: Below average; Orange: above average). The map was generated using DNAPlotter from Artemis (Carver et al., 2012).
Graphical circular map of Enterococcus faecalis strain mandacaium plasmid 1 (20,406 bp). From outer circle to the center: all CDS on forward and reverse strands; all CDS on forward strand; all CDS on reverse strand. Green: > 95% of identity with reference genome; Yellow: 70% to 95% of identity with reference genome; Red: < 70% of identity with reference genome. Inner circle shows GC content (Blue: Below average; Orange: above average). The map was generated using DNAPlotter from Artemis (Carver et al., 2012).

Figure 7:

Graphical circular map of Enterococcus faecalis strain mandacaium plasmid 2 (74,988 bp). From outer circle to the center: all CDS on forward and reverse strands; all CDS on forward strand; all CDS on reverse strand. Green: > 95% of identity with reference genome; Yellow: 70% to 95% of identity with reference genome; Red: < 70% of identity with reference genome. Inner circle shows GC content (Blue: Below average; Orange: above average). The map was generated using DNAPlotter from Artemis (Carver et al., 2012).
Graphical circular map of Enterococcus faecalis strain mandacaium plasmid 2 (74,988 bp). From outer circle to the center: all CDS on forward and reverse strands; all CDS on forward strand; all CDS on reverse strand. Green: > 95% of identity with reference genome; Yellow: 70% to 95% of identity with reference genome; Red: < 70% of identity with reference genome. Inner circle shows GC content (Blue: Below average; Orange: above average). The map was generated using DNAPlotter from Artemis (Carver et al., 2012).

Figure 8:

(A) RRE-containing region with a core biosynthetic gene encoding a sequence similar to the molecule Microcin 07 from Escherichia coli. (B) RiPP-like region with a core biosynthetic gene encoding a sequence similar to the Linocin M18. (C) Cyclic-lactone-autoinducer region with a core biosynthetic gene encoding a sequence similar to the molecule Enterocin NKR-5-3B from Enterococcus faecium.
(A) RRE-containing region with a core biosynthetic gene encoding a sequence similar to the molecule Microcin 07 from Escherichia coli. (B) RiPP-like region with a core biosynthetic gene encoding a sequence similar to the Linocin M18. (C) Cyclic-lactone-autoinducer region with a core biosynthetic gene encoding a sequence similar to the molecule Enterocin NKR-5-3B from Enterococcus faecium.

Figure 9:

Heat map of the Average Nucleotide Identity (ANI) analysis performed with Enterococcus faecalis strain mandacaium and Enterococcus spp. genomes deposited in the NCBI database. The genomic distance was calculated on the scale of 100 ANI. A distance of 0 (lightest color) corresponds to 100% ANI, indicating maximum similarity between genomes, while a distance of 100 (darkest color) corresponds to 0% ANI, indicating no similarity. Intermediate colors reflect proportional distances based on ANI values.
Heat map of the Average Nucleotide Identity (ANI) analysis performed with Enterococcus faecalis strain mandacaium and Enterococcus spp. genomes deposited in the NCBI database. The genomic distance was calculated on the scale of 100 ANI. A distance of 0 (lightest color) corresponds to 100% ANI, indicating maximum similarity between genomes, while a distance of 100 (darkest color) corresponds to 0% ANI, indicating no similarity. Intermediate colors reflect proportional distances based on ANI values.
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