Agave durangensis vinasse as a biocide for forest pest control

Abstract

In forestry practice, the prevention and fight against insect pests and diseases is a priority to preserve the health of these ecosystems. To combat it, insecticides of chemical and biological origin have been used. However, there are alternatives that have not yet been investigated, e.g., the use of agro-industrial waste. In the mezcal distillation process, polluting liquid residues called vinasses are generated, and these can be considered for pest control. In this project, the effect of vinasse from Agave durangensis subjected to different treatments was studied to evaluate its effect on forest phytopathogenic fungi. The mezcal vinasse was characterized physicochemically and by its metabolites. Furthermore, the percentage of inhibition in vitro of phytopathogenic (causing root wilt) fungi isolated from Pinus cooperi seedlings was studied. The fungi inhibition was related to the vinasse concentration. The lower pH and sterile raw vinasse showed a better inhibition effect. Four phytopathogenic strains of Pinus cooperi were isolated and identified, which corresponded to the genera Fusarium, Aspergillus, and Penicillium. None of the isolated were able to grow in potato dextrose-mezcal vinasse medium (PDMVM). Therefore, the mezcal vinasse showed fungicide activity in vitro against all strains.

Full Article

Agave durangensis Vinasse as a Biocide for Forest Pest Control

Ana M. Bailón-Salas,^a,δ Luis A. Ordaz-Diaz,^b and Pedro A. Domínguez-Calleros ^a,*

In forestry practice, the prevention and fight against insect pests and diseases is a priority to preserve the health of these ecosystems. To combat it, insecticides of chemical and biological origin have been used. However, there are alternatives that have not yet been investigated, e.g., the use of agro-industrial waste. In the mezcal distillation process, polluting liquid residues called vinasses are generated, and these can be considered for pest control. In this project, the effect of vinasse from Agave durangensis subjected to different treatments was studied to evaluate its effect on forest phytopathogenic fungi. The mezcal vinasse was characterized physicochemically and by its metabolites. Furthermore, the percentage of inhibition in vitro of phytopathogenic (causing root wilt) fungi isolated from Pinus cooperi seedlings was studied. The fungi inhibition was related to the vinasse concentration. The lower pH and sterile raw vinasse showed a better inhibition effect. Four phytopathogenic strains of Pinus cooperi were isolated and identified, which corresponded to the genera Fusarium, Aspergillus, and Penicillium. None of the isolated were able to grow in potato dextrose-mezcal vinasse medium (PDMVM). Therefore, the mezcal vinasse showed fungicide activity in vitro against all strains.

DOI: 10.15376/biores.17.1.1285-1300

Keywords: Mezcal; Metabolites sources; Fungicide activity

Contact information: a: Facultad de Ciencias Forestales, Doctorado Institucional en Ciencias Agropecuarias y Forestales. Río Papaloapan, Valle del Sur, Durango 34120 México; b: Ingeniería en Tecnología Ambiental, Universidad Politécnica de Durango, Carr. Dgo-Mex Km 9.5, Col. Dolores Hidalgo, Durango, Dgo 34300 México;

*Corresponding author: pdomingc@hotmail.com; δ postdoctoral student;

INTRODUCTION

Forest ecosystems provide various environmental services, e.g., clean air and water, conservation of biodiversity, and mitigation against the effects of climate change (MacDicken et al. 2016). However, these have been affected by various natural disturbances, e.g., pests (Montagné-Huck and Brunette 2018). There are many concerns about this because pests have an economic impact five times greater than forest fires (Logan et al. 2003). In addition, pathogens and pests have a high rate of adaptation, and climate change may favor outbreaks as well as their spread (Rubin-Aguirre et al. 2015; Wingfield et al. 2015; Jactel et al. 2019). In addition, the level of economic loss due to injuries caused by insects and forest pests is multifactorial, since they depend on the type of crop, temporal, and spatial location (Capinera 2020). Zhang et al. (2019) mention that diseases and pests endanger forestry. As such, the fight against insect pests and forest diseases is a priority, i.e., pink mealybug (Hibiscus mealybug), fungus, and debarker insects (Deschamps-Ramírez 2016).

For pest control, methyl bromide was widely used as a broad-spectrum soil fumigant until 2005 when it was banned (Rai 2020). As such, neonicotinoid pesticides have become the most widely used class of insecticides in the world (Simon-Delso et al. 2014). However, they produce considerable environmental impact (Saeed et al. 2019). Since most insecticides can be lethal to non-targeted organisms, this leads to the search for chemical and non-chemical alternatives to control pests (Simon-Delso et al. 2014). There are various sources of biomolecules, e.g., wastewater; however, the compounds present in wastewater have not been studied in-depth, since reducing pollution is the primary objective (Larif et al. 2015). Some residual waters from lignocellulosic biomass, liquid wastes of alcoholic fermentation called vinasses, specifically, are an alternative for preventing and controlling pests. The first reports of the use of vinasse for the control of phytopathogenic fungi dates back to 2008. In addition, it has been reported that wine vinasse showed an efficacy of 100% in terms of suppressing the growth of phytopathogenic fungi (Santos et al. 2008). Subsequently, sugar beet vinasse was tested for the control of nematodes in pepper crops as an alternative for disinfecting soil-borne pathogens (Núñez-Zofío et al. 2013). Due to this potential, vinasse should be studied as a biocide for the prevention and control of forest pests (Bailón-Salas et al. 2021). The raw material for making the biocide is available in Mexico. In addition, the Consejo Mexicano para la Regulación de la Calidad del Mezcal A.C. (COMERCAM) reports that the production of Mezcal in Mexico (2018 to 2019) increased by 40% (COMERCAM 2020). Therefore, its waste or by-products have also increased; in 2019 approximately 107 million liters of vinasse were generated. The vinasses contain various compounds, e.g., alcohols, aldehydes, phenols, and acids (Couallier et al. 2006; Freitas et al. 2018; Fuess et al. 2018). These compounds can participate or assist in the process of the inhibition or control of pests (Santos et al. 2008; Núñez-Zofío et al. 2013).

It is time to change this paradigm, stop seeing vinasse supplies only as waste and treating them as by-products, as well as revaluing the “waste” and giving them another type of value via the sustainable management of these materials (Ordaz-Díaz et al. 2019).

In addition, Mexico ranks tenth worldwide in total forest area. The state of Durango is the entity with the highest forest production in the country (SRNyMA 2006; SRNyMA 2015), whose most prominent genera are Pinus located in the coniferous forest (SRNyMA 2015). This genus is threatened by pine wilt disease (PWD), which is responsible for environmental and economic losses (Proença et al. 2017). Since fungal infection can cause the transplanted young pine seedlings to fail to establish, alternatives are needed to fumigate the soil substrate and control the disease (Gordon et al. 2015).

In this study the objectives were to determine the effect of the metabolites and microbial biomass contained in mezcal vinasse on fungal strains related to forest pests.

EXPERIMENTAL

Sampling and Physicochemical Characterization

The raw vinasse samples were taken from a mezcal factory in Nombre de Dios, Durango, collected in high-density polyethylene containers, and conserved at a temperature of 4 °C (Fig. S1a). The digested vinasse was taken from an anaerobic digester at the Universidad Politecnica de Durango (Fig. S1b).

Several parameters were used to characterize the mezcal vinasses; the total dissolved solids (TDS), electrical conductivity (EC), and pH were determined in situ with an HQ40d portable device (Hach Company, Loveland, CO). The settling solids (SS) were analyzed gravimetrically. The biochemical oxygen demand (BOD₅) was determined via the manometric-respirometric method with a BODTrak ™ II apparatus (Hach Company, Loveland, CO). Measurement of the chemical oxygen demand (COD) was determined using the closed reflux method (colorimetric), and the turbidity was determined using the spectrophotometric method (Hach DR 5000 Spectrophotometer), to analyze the water and wastewater (Rice et al. 2012). Different analytes, i.e., Cd, Fe, Ca, K, Mg, Na, Cr, Cu, As, Pb, Ni, and Zn, were analyzed via atomic absorption spectrometry (EAA) (AAnalyst^™, PerkinElmer, Waltham, MA) according to standard NMX-AA-051 (2016). All determinations were carried out in triplicate.

Metabolites Identification

The melanoidins quantification was carried out using an absorbance curve vs. the concentration of synthetic melanoidins obtained from the glucose-glycine model, with concentrations of 18, 2, 35, 47, 60, and 70 mg/L, via spectrophotometry at 340 nm. For the determination of phenols, the HI3864 aminoantipyrine method (Hanna Instruments) was determinate, in site immediately after sampling according to WPCF (2005).

Phytopathogenic Fungi Strain Isolation and Identification

Young Pinus cooperi seedlings samples at the nursery that presented fungal disease were collected (Fig. S3a through S3c). Soil, root, trunk, and foliage samples of P. cooperi were sown on PDA agar (Difco ^TM) (Fig. 3Sd through 3Sg) and incubated at a temperature of 28 °C ± 0.5 °C. The seedlings were donated by the Facultad de Ciencias Forestales y Ambientales nursery of the State of Durango. Morphological identification was made according to the methodology outlined by Leslie and Summerell (2008), Pitt and Hocking (2009), Houbraken et al. (2011), and Visagie et al. (2014). Colonies were visibly differentiated taking into consideration their shape, colony margins, color, and surface characteristics.

Bioassays

A multifactorial experimental design with multiple levels was carried out. The selected factors were as follows: type, concentration, sterile (yes/no), and pH with levels of 3, 5, 2, and 2, respectively (Table 1). The response variable was the diameter of the inhibition zone (mm).

Table 1. Factors and Levels Analyzed for the Vinasse Samples Against Isolated Agave durangensis Fungus

For the development of the treatment, the pH of the vinasse needed to be adjusted using calcium hydroxide, and the concentrations were diluted in distilled water. The characterization of some parameters, i.e., the pH, TDS, and EC, are shown in Table 2.

The in vitro inhibition effect of A. durangensis vinasse against fungal strains was analyzed because the main problem found in the studied nursery was root wilt. After the puncture inoculation and growing of the isolated fungal colonies from the root in the plates, 100 µL of mezcal vinasse was added and after incubation for 24 h, the zone of inhibition was measured through diameter inhibition zone (mm).

Table 2. Physicochemical Characterization of the Raw Stillage at a Neutral pH (adjusted) and the Anaerobically Digested Vinasse

Another experiment was performed in which the inhibition of the soil, trunk, and foliage isolates was evaluated using potato dextrose-mezcal vinasse medium (PDMVM). The PDMVM contained PDA agar (Difco^TM) dissolved in 1 L of mezcal vinasse. The plates were incubated at a temperature of 28 °C ± 0.5 °C up to 24 h and 48 h. Afterward, the growth was observed in comparison with the control. All experiments were performed in triplicate.

RESULTS AND DISCUSSION

Physicochemical Characterization

The physicochemical and metabolites characterization is shown in Table S2 and metal and metalloid characterization is shown in Table S1 (see Appendix). Mezcal vinasse is safe in terms of not generating additional problems due to contamination by heavy metals and metalloids. According to the results shown in Table S1, mezcal vinasse is free of various heavy metals, e.g., chromium, cadmium, lead, and nickel, and in addition, arsenic was not detected. Furthermore, mezcal vinasse is rich in calcium (575.33 mg/L), iron (15.04 mg/L), and magnesium (96.58 mg/L) (Table S1). Calcium in the soil contributes to the stabilization of organic carbon (Wan et al. 2021). In addition, it is widely known that the union between iron and organic carbon in the soil allows for its stabilization and storage (Yu et al. 2017). It has been shown that magnesium fertilizers can improve soil quality, specifically red soil (Nan et al. 2006). In addition, magnesium increases the availability of N and P (Velescu et al. 2021). Mezcal vinasse shows a copper concentration of 358.6 mg/L (Table S1). The presence of this ion is very positive for fungal control since it is a component of some biocides that protect wood from fungal attack (Schilling and Inda 2011). It is also part of commercial pesticide formulations applied in agriculture (Willis et al. 2016).

A characteristic low pH of 5.90 ± 0.05 was reported. An EC of 2.40 mS/cm ± 0.02 mS/cm was found, which was near the value reported by Mejía-Rivas et al. (2021). The color (4.355 ± 63.64 (Pt-Co) and turbidity (357 ± 24.88) NTU, were both related to the presence of melanoidins and polyphenols (Fitzgibbon et al. 1995). These results were also similar to those of Mejía-Rivas et al. (2021) in terms of A. durangensis vinasse. The biodegradable organic material fraction was very low, with a BOD₅ of 719 mg/L ± 4.24 mg/L and a COD of 19410 mg/L ± 70.71 mg/L. This left a high concentration of non-biodegradable organic matter.

Metabolites Identification

Figure S2 shows the curve vs. the concentration of synthetic melanoidins. The equation shows an adjustment coefficient of 0.99. The melanoidin concentration was 473.75 mg/L ±14.50 mg/L, while the phenol concentration was 1.30 mg/L ± 0.08 mg/L (as shown in Table S2). Phenolic compounds can inhibit microorganisms and are the responsible for the mortality of some pests (Larif et al. 2013; Freitas at al. 2018). It is suggested in the future to investigate and identify biologically active ingredients with a potential fungicidal or fungistatic effect.

Phytopathogenic Fungi Strain Isolation and Identification

Four strains were isolated in total; 1 from the root, 1 from the trunk, 2 from the foliage, and 3 from the soil. The isolates correspond to the Fusarium, Penicillium, and Aspergillus genera (Fig. 1). The fungal strain isolated from the root showed macroconidia and hyphal coils. Furthermore, white cottony aerial mycelium was present (Fig. 1). According to Leslie and Summerell (2008), this strain corresponds to the specie Fusarium circinatum. This pathogen has been reported to affect Pinus species and some other conifers in Mexico. Fusarium circinatum is one of the most important pathogens of Pinus spp., as it causes pitch canker (Wingfield et al. 2008).

Fig. 1. The identified strains: a) Fusarium circinatum; b) Aspergillus sp.; c) Penicillium sp.; and d) Aspergillus sp.

Table 3. Morphological Characterization of the Isolate Strain in PDA Culture Media

These three genera (Fig. 1) are of importance, since they are pathogens that decrease the viability of Pinus cooperi seedlings. Furthermore, the Aspergillus genus is the second most pathogenic fungus that affects the germination of Pinus seeds, followed by Fusarium (Ishtiaq et al. 2015). In addition, the three fungi isolated are fungi that have been found in the seeds of Pinus species and all cause a decrease in quality (López et al. 2021). Considering what was reported by other authors, it was decided to carry out the tests only on the fungus Fusarium circinatum, considered the most pathogenic for the selected pine species.

Bioassays

The inhibition percentage determination analysis is shown in Fig. 2. Fusarium circinatum was inhibited by raw vinasse treatment at a pH of 5.9. It did not show inhibition with the other three treatments, i.e., raw vinasse with neutral pH, dry vinasse, and digested vinasse. This can be attributed to the fact that the raw vinasse with neutral pH was adjusted with calcium hydroxide, which precipitated the compounds that are involved in the inhibition. Calcium hydroxide exerts the precipitation of melanoidins in A. durangensis vinasse (Mejía-Rivas et al. 2021). As for the dry vinasse, this may be due to the evaporation of all the phenolic compounds present in the sample, leaving only the effect of the melanoidins. Therefore, the melanoidins present in the mezcal vinasse do not have a fungicidal effect. Only coffee melanoidins have been reported to have a bactericidal effect (Rufián-Henares and Cueva 2009). In addition, the digested vinasse has been anaerobically biotransformed and no longer contains the inhibitory compounds to the fungus.

Fig. 2. In vitro control effect of F. circinatum with 100 µL at 24 h of 1) raw vinasse at a pH of 5.9; 2) raw vinasse at a neutral pH; 3) dry vinasse; and 4) digested vinasse

The effect of the mezcal vinasse concentration against fungal strain showed that the higher the concentration, the greater the inhibition (Fig. 3). However, this was observed at concentrations greater than 75%.

Fig. 3. In vitro control effect of F. circinatum with 100 µL at 24 h at different concentrations of mezcal vinasse

The sterile raw vinasse showed a better inhibition effect against Fusarium. This can be attributed to the fact that the sterile vinasse does not contain active microbial biomass that can grow in the medium (Fig. 4).

Fig. 4. In vitro control effect of F. circinatum using sterile raw vinasse and non-sterile vinasse

None of the four isolated fungi were able to grow in PDMVM (Table S3). Therefore, vinasse could be used as an alternative to heat treatment in commercial nurseries, as suggested by Berbegal et al. (2015).

CONCLUSIONS

1. Four fungi were identified in Pinus cooperi, which correspond to the genera Fusarium, Penicillium sp., and Aspergillus. It was decided to carry out the tests only on the fungus Fusarium circinatum, considered the most pathogenic for the selected pine species.
2. The percentage of inhibition of the fungus Fusarium circinatum is directly proportional to the vinasse concentration.
3. A lower pH favors fungi inhibition. It is necessary to consider the effect of long-term stillage application on soil and plants.
4. Sterile raw vinasse showed a better inhibition effect. All isolates were unable to grow on potato dextrose-mezcal vinasse medium (PDMVM), which was supplemented with vinasse.
5. The melanoidins present in the mezcal vinasse do not have a fungicidal effect.

ACKNOWLEDGMENTS

The support of the Consejo Nacional de Ciencia y Tecnologia (CONACyT) is gratefully appreciated for the support of the Sistema Nacional de Investigadores (SNI) as well as the postdoctoral scholarship (130780/20) received by one of the writers.

REFERENCES CITED

Bailón-Salas, A. M., Ordaz-Diaz, L. A., López-Serrano, P. M., Flores-Villegas, M. Y., and Dominguez-Calleros, P. A. (2021). “Wastewater as a resource for pest control: An overview,” BioResources 16(3), 6401-6425. DOI: 10.15376/biores.16.3.Bailon-Salas

Berbegal, M., Landeras, E., Sánchez, D., Abad‐Campos, P., Pérez‐Sierra, A., and Armengol, J. (2015). “Evaluation of Pinus radiata seed treatments to control Fusarium circinatum: Effects on seed emergence and disease incidence,” Forest Pathology 45(6), 525-533. DOI: 10.1111/efp.12204

Capinera, J. L. (2020). Handbook of Vegetable Pests, Academic Press, Cambridge, MA.

COMERCAM (2020). “Consejo Mexicano regulador del mezcal. Informe estadístico 2020,” (https://www.amma.org.mx/PDF/INF_ACTIVIDADES/INFORME2020.pdf), Accessed 24 August 2021.

Deschamps-Ramírez, P. (2016). “Plagas forestales: Hacia una política pública que fomente la acción de las comunidades dueñas de los bosques,” CCMSS, México, D.F. (https://www.ccmss.org.mx/wp-content/uploads/2016/05/CCMSS_Plagas_Forestales_Final.pdf, Accessed 20 December 2021.

Couallier, E. M., Ruiz, B. S., Lameloise, M.-L., and Decloux, M. (2006). “Usefulness of reverse osmosis in the treatment of condensates arising from the concentration of distillery vinasses,” Desalination 196(1-3), 306-317. DOI: 10.1016/j.desal.2006.02.002

Fitzgibbon, F. J., Nigam, P., Singh, D., and Marchant, R. (1995). “Biological treatment of distillery waste for pollution‐remediation,” Journal of Basic Microbiology 35(5), 293-301. DOI: 10.1002/jobm.3620350504

Freitas, P. V., Silva, D. R. d., Beluomini, M. A., Silva, J. L. d., and Stradiotto, N. R. (2018). “Determination of phenolic acids in sugarcane vinasse by HPLC with pulse amperometry,” Journal of Analytical Methods in Chemistry 2018, 1-10 DOI: 10.1155/2018/4869487

Fuess, L. T., Garcia, M. L., and Zaiat, M. (2018). “Seasonal characterization of sugarcane vinasse: Assessing environmental impacts from fertirrigation and the bioenergy recovery potential through biodigestion,” Science of the Total Environment 634, 29-40. DOI: 10.1016/j.scitotenv.2018.03.326

Gordon, T. R., Swett, C. L., and Wingfield, M. J. (2015). “Management of Fusarium diseases affecting conifers,” Crop Protection 73, 28-39. DOI: 10.1016/j.cropro.2015.02.018

Houbraken, J., Frisvad, J. C., and Samson, R. A. (2011). “Taxonomy of Penicillium section Citrina,” Studies in Mycology 70, 53-138. DOI: 10.3114/sim.2011.70.02

Ishtiaq, M., Noreen, M., Maqbool, M., Hussain, T., and Azam, S. (2015). “Analysis of fungal diversity impacts on Pinus roxburghaii seeds from pine forest and plant nurseries of Azad Kashmir, Pakistan,” Pakistan Journal of Botany 47(4), 1407-1414.

Jactel, H., Koricheva, J., and Castagneyrol, B. (2019). “Responses of forest insect pests to climate change: Not so simple,” Current Opinion in Insect Science 35, 103-108. DOI: 10.1016/j.cois.2019.07.010

Larif, M., Ouhssine, M., Soulaymani, A., and Elmidaoui, A. (2015). “Potential effluent oil mills and antibacterial activity polyphenols against some pathogenic strains,” Research on Chemical Intermediates 41(2), 1213-1225. DOI: 10.1007/s11164-013-1267-0

Larif, M., Zarrouk, A., Soulaymani, A., and Elmidaoui, A. (2013). “New innovation in order to recover the polyphenols of olive mill wastewater extracts for use as a biopesticide against the Euphyllura olivina and Aphis citricola,” Research on Chemical Intermediates 39(9), 4303-4313. DOI: 10.1007/s11164-012-0947-5

Leslie, J. F., and Summerell, B. A. (2008). The Fusarium Laboratory Manual, John Wiley & Sons, Hoboken, NJ.

Logan, J. A., Régnière, J., and Powell, J. A. (2003). “Assessing the impacts of global warming on forest pest dynamics,” Frontiers in Ecology and the Environment 1(3), 130-137. DOI: 10.1890/1540-9295(2003)001[0130:ATIOGW]2.0.CO;2

López, M. S. V., Badillo, M. E. V., Bautista, A. A., and Mancera Rico, A. (2021). “Seed fungi of Pinus montezumae Lamb. and Pinus greggii Engelm. ex Parl. stored under two relative humidities,” Revista Mexicana de Ciencias Forestales 12(66), 165-180. DOI: 10.29298/rmcf.v12i66.689

MacDicken, K., Jonsson, Ö., Piña, L., Marklund, L., Maulo, S., Contessa, V., Adikari, Y., Garzuglia, M., Lindquist, E., Reams, G., etc. (2016). Evaluación de los Recursos Forestales Mundiales 2015: ¿Cómo Están Cambiando los Bosques del Mundo? [Global Forest Resources Assessment 2015. How are the World’s Forests Changing?],” Food and Agriculture Organization, Rome, Italy.

Mejía-Rivas, C. A., Bailón-Salas, A. M., De la Peña-Arellano, L. A., Luis, A., Rodríguez-Rosales, M. D. J., and Ordaz-Díaz, L. A. (2021). “Evaluation of Opuntia ficus-indica potential as a natural coadjuvant for vinasse treatment,” BioResources 16(3), 6031-6056. DOI: 10.15376/biores.16.3.6031-6056

Montagné-Huck, C., and Brunette, M. (2018). “A bibliographic database on economic analysis of natural forest disturbances,” Data in Brief 20, 662-666. DOI: 10.1016/j.dib.2018.08.128

NMX-AA-051 (2001). “Water analysis – Determination of metals by atomic absorption in natural, drinking, wastewaters and wastewaters treated – Test method,” Secretaría de Economía, Mexico City, Mexico.

Núñez-Zofío, M., Larregla, S., Garbisu, C., Guerrero, M. M., Lacasa, C. M., and Lacasa, A. (2013). “Application of sugar beet vinasse followed by solarization reduces the incidence of Meloidogyne incognita in pepper crops while improving soil quality,” Phytoparasitica 41(2), 181-191. DOI: 10.1007%2Fs12600-012-0277-6

Ordaz-Diaz, L. A., Ceniceros, O. F. R., Rosales, M. D. J. R., Valle-Cervantes, S., Palacio, M. M. d., Kopa, K., and Bailón-Salas, A. M. (2019). “La vinaza del mezcal, un producto que de contaminante puede ser un fertilizante [Mezcal vinasse, a pollutant product that can be a fertilizer ],” Eek’ Revista de Divulgación Cientifica del COZCyT 8(4), 7-9.

Pitt, J. I., and Hocking, A. D. (2009). “Penicillium and related genera,” in: Fungi and Food Spoilage, J. I. Pitt, and A. D. Hocking (ed.), Springer, Boston, MA, pp. 169-273.

Proença, D. N., Grass, G., and Morais, P. V. (2017). “Understanding pine wilt disease: Roles of the pine endophytic bacteria and of the bacteria carried by the disease‐causing pinewood nematode,” MicrobiologyOpen 6(2), 1-20. DOI: 10.1002/mbo3.415

Rai, M., Abd-Elsalam, K. A., and Ingle, A. P. (2020). Pythium: Diagnosis, Diseases and Management, CRC Press, Boca Raton, FL.

Rubin-Aguirre, A., Sáenz-Romero, C., Lindig-Cisneros, R., del-Rio-Mora, A. A., Tena-Morelos, C. A., Campos-Bolaños, R., and del-Val, E. (2015). “Bark beetle pests in an altitudinal gradient of a Mexican managed forest,” Forest Ecology and Management 343, 73-79. DOI: 10.1016/j.foreco.2015.01.028

Rufián-Henares, J. A., and Cueva, S. P. d. l. (2009). “Antimicrobial activity of coffee melanoidins – A study of their metal-chelating properties,” Journal of Agricultural and Food Chemistry 57(2), 432-438. DOI: 10.1021/jf8027842

Saeed, Q., Ahmad, F., Iqbal, N., and Zaka, S. M. (2019). “Chemical control of polyphagous pests on their auxiliary hosts can minimize insecticide resistance: a case study of Spodoptera exigua Hübner (Lepidoptera: Noctuidae) in cotton agroecosystem,” Ecotoxicology and Environmental Safety 171, 721-727. DOI: 10.1016/j.ecoenv.2019.01.038

Santos, M., Diánez, F., Cara, M. d., and Tello, J. C. (2008). “Possibilities of the use of vinasses in the control of fungi phytopathogens,” Bioresource Technology 99(18), 9040-9043. DOI: 10.1016/j.biortech.2008.04.032

Schilling, J. S., and Inda, J. (2011). “Assessing the relative bioavailability of copper to fungi degrading treated wood,” International Biodeterioration & Biodegradation 65(1), 18-22. DOI: 10.1016/j.ibiod.2010.05.013

Simon-Delso, N., Amaral-Rogers, V., Belzenuces, L. P., Bonmatin, J. M., Chagnon, M., Downs, C., Furlan, L., Gibbons, D. W., Giorio, C., Girolami, V., et al. (2015). “Systemic insecticides (neonicotinoids and fipronil): trends, uses, mode of action and metabolites,” Environmental Science and Pollution Research 22, 5-22. DOI: 10.1007/s11356-014-3470-y

SRNMA (2006). “Programa estratégico forestal 2030 para Durango,” (http://www.conafor.gob.mx:8080/documentos/docs/12/177Programa%20Estrat%c3%a9gico%20Forestal%20de%20Durango.pdf), Accessed 24 August 2021.

SRNyMA (2015). “Estudio de la cuenca de abastecimiento forestal “Otinapa” del estado de Durango,” (http://www.conafor.gob.mx:8080/documentos/docs/22/6379Otinapa%20del%20Estado%20de%20Durango.pdf), Accessed 24 August 2021.

Sun, N., Zeng, X., Li, J., Gao, J., and Wang, B. (2006). “Effects of magnesium compound with fertilizer on daylily (Hemerocallis citrina Baroni) growth and soil nutrients,” Agricultural Sciences in China 5(2), 123-129. DOI: 10.1016/S1671-2927(06)60029-1

Velescu, A., Homeier, J., Bendix, J., Valarezo, C., and Wilcke, W. (2021). “Response of water-bound fluxes of potassium, calcium, magnesium and sodium to nutrient additions in an Ecuadorian tropical montane forest,” Forest Ecology and Management 501, 1-14. DOI: 10.1016/j.foreco.2021.119661

Visagie, C. M., Houbraken, J., Frisvad, J. C., Hong, S.-B., Klaassen, C. H. W., Perrone, G., Seifert, K. A., Varga, J., Yaguchi, T., and Samson, R. A. (2014). “Identification and nomenclature of the genus Penicillium,” Studies in Mycology 78, 343-371. DOI: 10.1016/j.simyco.2014.09.001

Wan, D., Ma, M., Peng, N., Luo, X., Chen, W., Cai, P., Wu, L., Pan, H., Chen, J., Yu, G., et al. (2021). “Effects of long-term fertilization on calcium-associated soil organic carbon: Implications for C sequestration in agricultural soils,” Science of The Total Environment 772, 1-9. DOI: 10.1016/j.scitotenv.2021.145037

Willis, B. E., and Bishop, W. M. (2016). “Understanding fate and effects of copper pesticides in aquatic systems,” Journal of Geoscience and Environment Protection 4(5), 37-42. DOI: 10.4236/gep.2016.45004

Wingfield, M. J., Brockerhoff, E. G., Wingfield, B. D., and Slippers, B. (2015). “Planted forest health: The need for a global strategy,” Science 349(6250), 832-836. DOI: 10.1126/science.aac6674

Wingfield, M. J., Hammerbacher, A., Ganley, R. J., Steenkamp, E. T., Gordon, T. R., Wingfield, B. D., and Coutinho, T. A. (2008). “Pitch canker caused by Fusarium circinatum – A growing threat to pine plantations and forests worldwide,” Australasian Plant Pathology 37(4), 319-334. DOI: 10.1071/AP08036

WPCF, A. A. (2005). American Public Health Association American Water Works Association & Water Pollution Control Federation. Standard Methods for the examination of water and wastewater. Washington, DC (USA).

Yu, G., Xiao, J., Hu, S., Polizzotto, M. L., Zhao, F., McGrath, S. P., Li, H., Ran, W., and Shen, Q. (2017). “Mineral availability as a key regulator of soil carbon storage,” Environmental Science & Technology 51(9), 4960-4969. DOI: 10.1021/acs.est.7b00305

Zhang, J., Huang, Y., Pu, R., Gonzalez-Moreno, P., Yuan, L., Wu, K., and Huang, W. (2019). “Monitoring plant diseases and pests through remote sensing technology: A review,” Computers and Electronics in Agriculture 165, 1-14. DOI: 10.1016/j.compag.2019.104943

Article submitted: October 18, 2021; Peer review completed: December 11, 2021; Revised version received and accepted: December 20, 2021; Published: January 5, 2022.

DOI: 10.15376/biores.17.1.1285-1300

APPENDIX

Supplemental Material

Fig. S1. Sampling of the a) storage pit; and b) anaerobic digester of the mezcal vinasses

Table S1. Metal and Metalloid Characterization

Table S2. Physicochemical and Metabolites Characterization

Fig. S2. Absorbance vs concentration of the synthetic melanoidins

Fig. S3. Sampling of the infected young pine seedlings with isolated fungi Pinus cooperi: a) Healthy Pinus cooperi seedlings; b) Diseased Pinus cooperi seedlings; c) Root wilt; d) soil sample in PDA medium at 48 h of incubation; e) root sample in PDA medium at 24 h of incubation; and sample of the trunk root and aerial part in PDA medium at f) 24 h; and g) 48 h of incubation

Table S3. Evaluation of the Inhibitory Effect of A. durangensis Vinasse Against 4 Isolated Fungal Strains of P. cooperi at 24 h and 48 h of Incubation