<i>In vitro</i> antimicrobial activity of methanolic extracts from cutaneous secretions of Amazonian amphibians against phytopathogens of agricultural interest

RAASCH-FERNANDES, Livia Deice; BONALDO, Solange Maria; RODRIGUES, Domingos de Jesus; FERRARINI, Stela Regina; VERÇOSA, Ana Gabriela Araújo; OLIVEIRA, Daiane Lopes de

doi:10.1590/1809-4392201904462

Livia Deice RAASCH-FERNANDES

Universidade Federal de Mato Grosso (UFMT), Programa de Pós-Graduação em Ciências Ambientais (PPGCAM), Campus Sinop, Mato Grosso, Brazil

Solange Maria BONALDO ^** Corresponding author: sbonaldo@ufmt.br

Universidade Federal de Mato Grosso (UFMT), Programa de Pós-Graduação em Ciências Ambientais (PPGCAM), Campus Sinop, Mato Grosso, Brazil

Universidade Federal de Mato Grosso (UFMT), Instituto de Ciências Agrárias e Ambientais (ICAA), Campus Sinop, Mato Grosso, Brazil

http://orcid.org/0000-0002-2240-2700

Domingos de Jesus RODRIGUES

Universidade Federal de Mato Grosso (UFMT), Programa de Pós-Graduação em Ciências Ambientais (PPGCAM), Campus Sinop, Mato Grosso, Brazil

Stela Regina FERRARINI

Universidade Federal de Mato Grosso (UFMT), Instituto de Ciências da Saúde, Sinop, Mato Grosso, Brazil

Ana Gabriela Araújo VERÇOSA

Universidade Federal de Mato Grosso (UFMT), Instituto de Ciências Agrárias e Ambientais (ICAA), Campus Sinop, Mato Grosso, Brazil

Daiane Lopes de OLIVEIRA

Universidade Federal de Mato Grosso (UFMT), Instituto de Ciências Agrárias e Ambientais (ICAA), Campus Sinop, Mato Grosso, Brazil

* Corresponding author: sbonaldo@ufmt.br

Associate Editor:

Jorge Maurício David

Figures (2)
Tables (4)

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Figure 1
Growth of mycelial colonies of the pathogens Fusarium udum, Fusarium solani, Colletotrichum truncatum, Aspergillus flavus, Rhizoctonia solani, Macrophomina phaseolina, and Calonectria pseudometrosideri treated with the methanolic extract of Rhaebo guttatus cutaneous secretion. A − negative control (sterile water); B − positive control (fungicide); C − extract concentration 0.1 mg mL^-¹; D − 0.2 mg mL^-¹; E − 0.3 mg mL^-¹; F − 0.4 mg mL^-¹; G − 0.5 mg mL^-¹. This figure is in color in the electronic version.

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Figure 2
Growth of the mycelial colonies of the pathogens Fusarium udum, Fusarium solani, Colletotrichum truncatum, Aspergillus flavus, Rhizoctonia solani, Macrophomina phaseolina, and Calonectria pseudometrosideri treated with the methanolic extract of Rhinella marina cutaneous secretion. A − negative control (sterile water); B − positive control (fungicide); C − extract concentration 0.1 mg mL^-¹; D − 0.2 mg mL^-¹; E − 0.3 mg mL^-¹; F − 0.4 mg mL^-¹; G − 0.5 mg mL^-¹. This figure is in color in the electronic version.

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Table 1
Mycelial growth (MG) and mycelial growth rate index (MGRI) of fungal pathogens treated with different concentrations of the methanolic extract of Rhaebo guttatus (Rg) and Rhinella marina (Rm) cutaneous secretions. C- = negative control (sterile water); C+ = positive control [fungicides pyraclostrobin + metconazole (F. udum, F. solani, C. truncatum, A. flavus, R. solani and M. phaseolina) or azoxystrobin + ciproconazole (C. pseudometrosideri)].

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Table 2
Percentage of mycelial growth inhibition (%MGI) of fungal pathogens treated with methanolic extracts of Rhaebo guttatus (Rg) and Rhinella marina (Rm) cutaneous secretions. C+ = positive control [fungicides pyraclostrobin + metconazole (F. udum, F. solani, C. truncatum, A. flavus, R. solani and M. phaseolina) or azoxystrobin + ciproconazole (C. pseudometrosideri)].

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Table 3
Percentage of sporulation inhibition (%SI) of fungal pathogens treated with methanolic extracts of Rhaebo guttatus (Rg) and Rhinella marina (Rm) cutaneous secretions. C+ = positive control [fungicides pyraclostrobin + metconazole (F. udum, F. solani, C. truncatum, A. flavus, R. solani and M. phaseolina) or azoxystrobin + ciproconazole (C. pseudometrosideri)].

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Table 4
Effect of different concentrations of the methanolic extracts of cutaneous secretions of Rhaebo guttatus (Rg) and Rhinella marina (Rm) on fungal pathogens measured by microsclerotia of Rhizoctonia solani, percentage of inhibition of appressorial formation (%AFI) in Colletotrichum truncatum, and percentage of inhibition of conidial germination (%CGI) of Fusarium udum, C. truncatum, Aspergillus flavus and Calonectria pseudometrosideri. C- = negative control (sterile water); C+ = positive control [fungicides pyraclostrobin + metconazole (R. solani, F. udum, C. truncatum and A. flavus) or azoxystrobin + ciproconazole (C. pseudometrosideri)].

Extract (mg mL-¹)	Fusarium udum		Fusarium solani		Colletotrichum truncatum		Aspergillus flavus		Rhizoctonia solani		Macrophomina phaseolina		Calonectria pseudometrosideri
Extract (mg mL-¹)	MG (cm)	MGRI (cm day^-1)	MG (cm)	MGRI (cm day^-1)	MG (cm)	MGRI (cm day^-1)	MG (cm)	MGRI (cm day^-1)	MG (cm)	MGRI (cm day^-1)	MG (cm)	MGRI (cm day^-1)	MG (cm)	MGRI (cm day^-1)
Rhaebo guttatus
C-	6.60 ± 0.22 c	0.94 ± 0.03 c	7.18 ± 0.52 c	0.90 ± 0.06 c	6.30 ± 0.77 a	0.53 ± 0.06 a	6.78 ± 0.49 c	0.75 ± 0.05 c	6.48 ± 2.83 b	2.16 ± 0.94 b	7.94 ± 0.22 d	2.65 ± 0.07 c	5.88 ± 0.98 a	0.42 ± 0.07 a
C+	1.37 ± 0.22 a	0.20 ± 0.03 a	3.56 ± 0.33 a	0.45 ± 0.04 a	6.52 ± 0.29 a	0.54 ± 0.02 a	2.53 ± 0.47 a	0.28 ± 0.05 a	2.48 ± 0.32 a	0.83 ± 0.11 a	1.07 ± 0.17 a	0.36 ± 0.06 a	5.00 ± 1.37 a	0.36 ± 0.10 a
0.1	6.62 ± 0.33c	0.95 ± 0.05 c	5.71 ± 0.62 b	0.71 ± 0.08 b	6.33 ± 0.40 a	0.53 ± 0.03 a	4.20 ± 0.70 b	0.47 ± 0.08 b	7.73 ± 1.08 b	2.58 ± 0.36 b	6.88 ± 0.39 b	2.29 ± 0.13 b	6.64 ± 1.39 b	0.47 ± 0.10 b
0.2	6.74 ± 0.42c	0.96 ± 0.06 c	5.96 ± 0.57 b	0.75 ± 0.07 b	6.60 ± 0.31 a	0.55 ± 0.03 a	5.99 ± 1.46 c	0.67 ± 0.16 c	8.06 ± 0.83 b	2.69 ± 0.28 b	7.53 ± 0.36 c	2.51 ± 0.12 c	7.36 ± 0.97 b	0.53 ± 0.07 b
0.3	6.82 ± 0.35 c	0.97 ± 0.05 c	5.48 ± 0.77 b	0.69 ± 0.10 b	6.37 ± 0.57 a	0.53 ± 0.05 a	6.02 ± 0.88 c	0.67 ± 0.10 c	7.64 ± 1.49 b	2.55 ± 0.50 b	7.76 ± 0.20 c	2.59 ± 0.07 c	5.04 ± 0.47 a	0.36 ± 0.03 a
0.4	6.84 ± 0.47 c	0.98 ± 0.07 c	5.62 ± 0.54 b	0.70 ± 0.07 b	6.74 ± 0.38 a	0.56 ± 0.03 a	5.67 ± 1.23 c	0.63 ± 0.14 c	8.03 ± 0.70 b	2.68 ± 0.23 b	7.92 ± 0.26 d	2.64 ± 0.09 c	5.23 ± 1.48 a	0.37 ± 0.11 a
0.5	5.21 ± 1.32 b	0.74 ± 0.19 b	5.84 ± 0.82 b	0.73 ± 0.10 b	6.41 ± 0.66 a	0.53 ± 0.05 a	4.13 ± 2.13 b	0.46 ± 0.24 b	8.19 ± 0.46 b	2.58 ± 0.31 b	8.10 ± 0.22 d	2.70 ± 0.07 c	4.92 ± 0.39 a	0.35 ± 0.03 a
Rhinella marina
C-	4.90 ± 1.78 b	0.61 ± 0.22 b	6.44 ± 0.32 b	0.81 ± 0.04 b	6.48 ± 0.37 a	0.54 ± 0.03 b	6.93 ± 1.47 b	0.63 ± 0.13 b	6.68 ± 0.38 b	3.34 ± 0.19 b	8.32 ± 0.33 b	2.77 ± 0.11 b	5.87 ± 1.28 a	0.31 ± 0.07 a
C+	2.70 ± 0.31 a	0.34 ± 0.04 a	3.26 ± 0.26 a	0.41 ± 0.03 a	5.94 ± 0.60 a	0.50 ± 0.05 a	3.87 ± 0.43 a	0.35 ± 0.04 a	2.23 ± 0.50 a	1.12 ± 0.25 a	1.60 ± 0.09 a	0.53 ± 0.03 a	6.83 ± 1.86 a	0.36 ± 0.10 a
0.1	5.73 ± 1.91 b	0.72 ± 0.24 b	5.93 ± 1.09 b	0.74 ± 0.14 b	6.82 ± 0.38 a	0.57 ± 0.03 b	7.06 ± 1.00 b	0.64 ± 0.09 b	6.66 ± 0.43 b	3.33 ± 0.22 b	8.10 ± 0.35 b	2.70 ± 0.12 b	7.21 ± 1.24 a	0.38 ± 0.07 a
0.2	6.77 ± 0.18 b	0.85 ± 0.02 b	5.66 ± 1.18 b	0.71 ± 0.15 b	6.74 ± 0.37 a	0.56 ± 0.03 b	6.74 ± 0.34 b	0.61 ± 0.03 b	7.08 ± 0.22 b	3.54 ± 0.11 b	8.44 ± 0.08 b	2.81 ± 0.03 b	6.14 ± 0.48 a	0.32 ± 0.03 a
0.3	6.10 ± 0.43 b	0.76 ± 0.05 b	6.01 ± 1.35 b	0.75 ± 0.17 b	6.76 ± 0.41 a	0.56 ± 0.03 b	7.31 ± 0.77 b	0.66 ± 0.07 b	6.62 ± 0.56 b	3.31 ± 0.28 b	8.13 ± 0.54 b	2.71 ± 0.18 b	7.01 ± 1.47 a	0.37 ± 0.08 a
0.4	6.41 ± 0.77 b	0.80 ± 0.10 b	5.03 ± 0.26 b	0.63 ± 0.03 b	6.86 ± 0.46 a	0.57 ± 0.04b	6.76 ± 1.08 b	0.62 ± 0.07 b	6.87 ± 0.13 b	3.44 ± 0.06 b	8.37 ± 0.14 b	2.79 ± 0.05 b	6.86 ± 0.86 a	0.36 ± 0.05 a
0.5	5.58 ± 1.75 b	0.70 ± 0.22 b	5.68 ± 0.42 b	0.71 ± 0.05 b	5.63 ± 0.81 a	0.47 ± 0.07 a	6.80 ± 0.87 b	0.62 ± 0.10 b	7.03 ± 0.27 b	3.52 ± 0.14 b	8.33 ± 0.18 b	2.78 ± 0.06 b	7.28 ± 1.39 a	0.38 ± 0.07 a

Extract (mg mL-¹)	Fusarium udum		Fusarium solani		Colletotrichum truncatum		Aspergillus flavus		Rhizoctonia solani		Macrophomyia phaseolina		Calonectria pseudometrosideri
Extract (mg mL-¹)	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm
C+	79.2 ± 3.1 a	38.2 ± 24.2 a	50.2 ± 5.9 a	49.2 ± 4.6 a	2.1 ± 3.1 a	9.4 ± 9.3 a	62.7 ± 6.1 a	41.1 ± 18.0 a	50.8 ± 33.3 a	66.5 ± 7.8 a	86.5 ± 2.1 a	80.7 ± 1.5 a	19.6 ± 19.7 a	4.2 ± 7.4 a
0.1	1.9 ± 3.0 c	12.1 ± 27.2 b	20.0 ± 11.2 b	10.9 ± 15.3 b	4.4 ± 5.9 a	0.6 ± 1.3 b	38.0 ± 9.7 a	10.6 ± 11.2 b	8.0 ± 13.3 b	1.7 ± 2.5 b	13.3 ± 4.8 b	2.6 ± 2.4 b	7.4 ± 16.6 b	0.5 ± 1.1 a
0.2	2.3 ± 5.1 c	1.4 ± 2.5 b	16.8 ± 12.4 b	13.8 ± 13.3 b	2.3 ± 5.2 a	0.0 ± 0.0 b	13.3 ± 18.6 b	8.1 ± 10.8 b	5.1 ± 9.7 b	0.0 ± 0.0 b	5.1 ± 5.0 c	0.4 ± 0.8 b	2.6 ± 5.9 b	4.6 ± 10.3 a
0.3	1.4 ± 2.8 c	6.5 ± 9.0 b	23.3 ± 12.9 b	11.9 ± 17.2 b	5.3 ± 7.7 a	2.2 ± 5.1 b	11.3 ± 12.4 b	5.6 ± 10.1 b	8.1 ± 18.2 b	4.9 ± 6.9 b	2.9 ± 2.7 c	2.3 ± 3.2 b	15.1 ± 9.5 a	0.8 ± 1.8 a
0.4	1.8 ± 3.7 c	5.6 ± 8.4 b	21.7 ± 3.4 b	21.8 ± 4.4 b	1.6 ± 3.3 a	0.3 ± 0.7 b	16.9 ± 13.8 b	11.7 ± 16.1 b	3.6 ± 8.2 b	1.1 ± 1.8 b	1.5 ± 3.5 d	1.1 ± 1.2 b	16.9 ± 18.6 a	4.7 ± 10.6 a
0.5	21.3 ± 18.2 b	3.6 ± 5.5 b	19.1 ± 13.3 b	11.6 ± 7.3 b	2.7 ± 4.9 a	13.1 ± 13.5 a	40.6 ± 27.9 a	7.2 ± 9.3 b	0.3 ± 0.8 b	0.0 ± 0.0 b	0.2 ± 0.5 d	0.8 ± 1.1 b	17.9 ± 13.7 a	1.5 ± 3.3 a

Extract (mg mL-¹)	Fusarium udum		Fusarium solani		Colletotrichum truncatum		Aspergillus flavus		Calonectria pseudometrosideri
Extract (mg mL-¹)	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm
C+	6.6 ± 14.9 a	84.6 ± 29.9 a	18.5 ± 40.7 a	53.1 ± 49.2 a	20.0 ± 44.7 a	40.0 ± 54.8 a	95.9 ± 2.2 a	85.7 ± 9.7 a	60.0 ± 55.8 a	39.1 ± 44.4 a
0.1	0.0 ± 0.0 a	77.3 ± 43.4 a	58.2 ± 41.7 a	35.2 ± 49.0 a	0.0 ± 0.0 a	20.0 ± 44.7 a	93.1 ± 7.0 a	35.8 ± 28.4 b	12.5 ± 28.0 a	49.9 ± 46.1 a
0.2	20.0 ± 44.7 a	71.8 ± 41.5 a	55.5 ± 50.8 a	17.4 ± 38.9 a	40.0 ± 54.7 a	27.6 ± 43.7 a	18.3 ± 31.9 b	17.5 ± 25.1 b	18.7 ± 41.9 a	31.7 ± 45.9 a
0.3	20.0 ± 44.7 a	79.0 ± 44.2 a	27.8 ± 34.2 a	29.7 ± 29.0 a	40.0 ± 54.7 a	40.0 ± 54.8 a	29.1 ± 41.4 b	11.1 ± 11.9 b	52.5 ± 50.3 a	26.8 ± 41.0 a
0.4	20.0 ± 44.7 a	72.1 ± 40.9 a	64.6 ± 36.2 a	15.6 ± 34.9 a	20.0 ± 44.7 a	18.4 ± 41.3 a	45.4 ± 35.7 b	54.9 ± 35.3 a	38.7 ± 53.1 a	40.5 ± 46.9 a
0.5	6.6 ± 14.9 a	66.3 ± 30.9 a	25.2 ± 34.7 a	32.4 ± 33.2 a	20.0 ± 44.7 a	40.0 ± 54.8 a	83.6 ± 26.3 a	42.1 ± 41.4 b	58.7 ± 53.7 a	38.3 ± 52.6 a

Extract (mg mL-¹)	Rhizoctonia solani		Fusarium udum		Colletotrichum truncatum				Aspergillus flavus		Calonectria pseudometrosideri
	Microsclerotia		%CGI		%CGI		%AFI		%CGI		%CGI
	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm	Rg	Rm
C-	4.0 ± 0.0 a	3.8 ± 0.4 b	-	-	-	-	-	-	-	-	-	-
C+	4.0 ± 0.0 a	3.2 ± 1.7 b	83.9 ± 16.0 a	93.5 ± 8.2 a	95.6 ± 4.1 a	95.4 ± 4.7 a	100.0 ± 0.0 a	100.0 ± 0.0 a	66.6 ± 4.8 a	72.0 ± 9.4 a	95.7 ± 4.9 a	91.6 ± 6.1 a
0.1	4.0 ± 0.0 a	4.0 ± 0.0 b	0.0 ± 0.0 c	67.7 ± 10.6 b	41.4 ± 2.2 b	55.9 ± 8.2 b	85.3 ± 15.3 b	100.0 ± 0.0 a	22.9 ± 6.9 b	8.0 ± 5.4 b	38.3 ± 3.5 c	18.1 ± 10.5 b
0.2	4.0 ± 0.0 a	2.4 ± 2.1 a	31.6 ± 24.4 b	66.0 ± 11.4 b	52.8 ± 12.1 b	42.6 ± 10.9 b	99.2 ± 2.2 a	98.2 ± 5.0 a	24.2 ± 15.7 b	11.0 ± 9.3 b	37.6 ± 2.8 c	28.9 ± 13.8 b
0.3	4.0 ± 0.0 a	1.0 ± 1.7 a	55.5 ± 19.4 a	59.3 ± 11.6 b	43.5 ± 9.3 b	41.5 ± 14.7 b	95.6 ± 6.1 a	95.1 ± 11.6 a	20.3 ± 23.1 b	7.1 ± 7.0 b	50.9 ± 3.2 b	27.5 ± 9.0 b
0.4	3.2 ± 1.7 a	4.0 ± 0.0 b	33.9 ± 25.6 b	57.1 ± 12.9 b	39.3 ± 16.0 b	42.4 ± 18.1 b	98.3 ± 3.2 a	98.2 ± 3.3 a	12.6 ± 5.5 b	6.4 ± 5.44 b	55.4 ± 2.4 b	27.0 ± 8.4 b
0.5	3.8 ± 0.4 a	4.0 ± 0.0 b	33.0 ± 25.5 b	53.0 ± 14.1 b	26.9 ± 21.0 c	21.6 ± 19.9 c	85.9 ± 13.1 b	63.1 ± 36.1 b	18.5 ± 7.7 b	11.1 ± 11.2 b	50.9 ± 2.2 b	30.6 ± 12.7 b

[1] * Corresponding author: sbonaldo@ufmt.br

Brasil

Brasil

In vitro antimicrobial activity of methanolic extracts from cutaneous secretions of Amazonian amphibians against phytopathogens of agricultural interest

Atividade antimicrobiana in vitro de extratos metanólicos de secreções cutâneas de anfíbios amazônicos sobre fitopatógenos de interesse agrícola

ABSTRACT