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Article

Evaluation of the Fungitoxic Effect of Extracts from the Bark of Quercus laeta Liebm, the Cob of Zea mays and the Leaves of Agave tequilana Weber Blue Variety against Trametes versicolor L. Ex Fr

by
Alberto Gálvez-Martínez
1,
Rosa María Jiménez-Amezcua
1,
José Anzaldo-Hernández
2,
María Guadalupe Lomelí-Ramírez
2,
José Antonio Silva-Guzmán
2,
José Guillermo Torres-Rendón
2,* and
Salvador García-Enriquez
2
1
Chemical Engineering Department, University of Guadalajara, Marcelino García Barragán Blvd. 1451, Guadalajara 44430, Mexico
2
Department of Wood, Cellulose and Paper, University of Guadalajara, Guadalajara-Nogales Highway 15.5 Km, Guadalajara 45220, Mexico
*
Author to whom correspondence should be addressed.
Forests 2024, 15(7), 1204; https://doi.org/10.3390/f15071204
Submission received: 8 June 2024 / Revised: 9 July 2024 / Accepted: 10 July 2024 / Published: 11 July 2024
(This article belongs to the Special Issue Wood Durability and Protection)
Browse Figures
Versions Notes

Abstract

:
Wood products used in outdoor applications can be degraded by xylophage organisms. For this reason, such products require treatments based on biocides in order to delay their service life. This brings troubles of its own due to the inherent toxicity of these treatments towards humans and the environment. Therefore, it is imperative to find less-toxic natural preservatives. In this context, this work deals with the evaluation of the fungitoxic effect of raw extracts obtained from three types of agroindustrial waste materials: bark of Quercus laeta spp., the cob of Zea mays, and the leaves of Agave tequilana Weber Blue variety. Extracts were incorporated into the test wood Alnus acuminata (Aile wood) via a full-cell process. Bark extracts provided excellent protection against the attack of Trametes versicolor (L. ex. Fr.) Pilát, improving the decay resistance of Aile wood from being nonresistant to resistant. Also, bark extracts from Q. laeta showed less leaching than the other extracts.

1. Introduction

Wood was most likely one of the first structural materials used by humans. Due to its organic nature, woods can be affected by several biological agents that can rot and destroy it under certain conditions. This susceptibility varies depending on the species and the conditions of the place where the wood is being used. For example, woods in places of low humidity are less prone to be damaged than woods that are directly in humid places or in direct contact with the soil. Fungi are the main rotting agents of wood; they cause great economic losses and drastically reduce the service life of woods [1,2].
The main biological degradation agents of wood are xylophage fungi. This type of fungus is important in nature due to their parasitic and recycling functions. Also, because of the mentioned degradation effect on wood, xylophage fungi have a critical economic importance in forest products [3,4].
In general, chemical treatments used to protect wood against xylophage fungi have inherent toxicity [5]. Harmful substances to humans and to the environment such as arsenic, chromium, copper and boron have been used as wood preservatives [6].
Therefore, it is necessary to find less-toxic and environmentally friendlier natural preservatives [7,8,9,10]. In this context, extracts from lignocellulosic sources with applicability as natural preservatives have been reported, such as those from teak heartwood [11], Ocotea lancifolia leaves [12], valonia, chestnut, tara, sulphited oak [13], scots pine knotwood, and black locust heartwood [14], among others.
On the other hand, solid waste generated by human activity is an important source of pollution. Thus, alternatives to exploit solid waste/byproducts are being explored [15,16]. In this sense, a circular economic approach, in which waste from an industry is utilized as a starting material in another industry, can be considered. The agriculture industry is one of the main generators of solid waste. Bark, stems, roots, leaves, and other parts of plants are not processed, thus becoming waste [17,18]. In the Mexican agroindustry, the bark of Quercus laeta, the cob of Zea mays, and the leaves of Agave tequilana Weber Blue variety are typical byproducts/waste [19,20,21], so their potential applications are worth exploring.
Quercus laeta Liebm have been used by man since ancient times to obtain wood, as a source of tannins, for tanning skins, and in medicine. These plants, also known as oaks or acorn trees, belong to the genus Quercus, one of the most important worldwide [22,23,24], which is of special interest in North America [25]. Also, it has been used as food for cows and goats [26] and as a component of wood–plastic composites [27,28]. On the other hand, the leaves of A. tequilana have been used for the extraction and quantification of fructans [29], fructo-oligosaccharides [30,31], inulin, and free sugars [32], as well as for the production of bioethanol [33,34,35,36], succinic acid [37], hyaluronic acid [38], and fibers for handmade paper [39,40] and crafts [41]. Fibers from A. tequilana weber have been extracted and intended to be used for food inclusivity [42] and to feed sheep [43]. They have also been used to obtain nanocellulose [44], for the biosorption of Pb II [45], and for preparation of biocomposites [46]. Finally, corn cob has been used extensively for several applications. For example, it has been used to obtain biochar [47,48]; as a food supplement for rabbits [49,50]; as a source of forage for cows [51]; as a biosorbent of dyes [52,53]; and in the production of cellulases, laccases [54], and edible mushrooms [55,56]. Also, it has been used in paper production [57] and for the preparation of biofilms [58]. This work deals with the preparation and characterization of extracts from these three waste materials. The main objective of this study is to demonstrate that these waste-derived extracts can be used as natural preservatives for wood against the attacks of xylophage fungi.

2. Materials and Methods

2.1. Materials

Barks of Quercus laeta Liebm were collected from the mountain range of Tequila, in Jalisco, Mexico. Cobs of Zea mays were provided by the Hermanos Esparza S.A. de C.V. company in Guadalajara, Mexico. Leaves of Agave tequilana Weber Blue variety were donated by the Bioing biomasas company in Tequila, Mexico. N-Hexane (97%) and ketone (99.9%) were purchased from Sigma-Aldrich (Toluca, Mexico). Malt extract Agar was purchased from BD Difco. Figure 1 shows the methodology followed in this work.

2.2. Preparation of Extracts

The starting materials (bark, corn cob, and agave leaves) were first dried at 60 °C for 48 h; then, they were grinded (blade mill Pulvex 95) and sifted. Particles that passed the 65-mesh sieve (297–211 µm) were collected for extraction.
Extractions were carried out by submerging the sifted agro-industrial residues (50 g per material) in a hexane/ketone/water solution (54:44:2) [7]. The mixing was stirred for 24 h at 20 °C. Then, undesired solids were removed by filtration and the solvent was partially evaporated using a rotary evaporator. The concentrated solution was placed in an oven at 40 °C until it was completely dry.

2.3. Wood Sample Preparation

Wood samples from the heartwood of Alnus acuminata (Aile wood) were collected in the San Juan Nuevo sawmill in Michoacan, Mexico. No damage by microorganisms was observed. Collected samples were cut into cubes of 19 mm on each side. Also, samples were cut into blocks of 12 × 35 × 3 mm3 to be used as feeding blocks for the rooting tests. The cubes were dried at 60 °C until constant weight (T1) and their dimensions were measured. The volume of each cube was also determined. The density of the cubes was obtained, and groups of cubes were formed depending on their density.

Impregnation

Cubes were impregnated using solutions with different concentrations of extracts (0.1, 1.0, 3.0 and 5.0 g/L). Solutions were prepared using the same solvent system utilized for extraction (hexane/ketone/water, 54:44:2). The number of impregnated cubes used for the leaching and bioassay tests are shown in Table 1.
Impregnation of cubes was carried out using the full-cell method. A vacuum–pressure–vacuum cycle was performed at 30 cm Hg (30 min), 5 kg/cm2 (60 min), and 30 cm Hg (25 min), respectively (Figure 2b). The equipment utilized to impregnate the cubes is represented in Figure 2. After completion of the process, the cubes were taken out from the biocide solution and weighted (recorded as weight T2). Then, the cubes were dried at 60 °C until achieving constant weight (recorded as weight T3).
Absorption is calculated using Equation (1):
A = T 2 − T 1 V ;   k g m 3
where V is the volume of impregnated wood.
Results were classified according to the following ranges: high absorption (HA), higher than 200 kg/m3; good absorption (GA), from 150 to 200 kg/m3; bad absorption (BA), from 100 to 150 kg/m3; and zero absorption (ZA), less than 100 kg/m3 [59].
Retention is the quantity of active toxic components in the preservative that remained in the wood after impregnation. It can be calculated using Equation (2).
R = G × C V   × 10 ;   k g m 3
where
G = mass of absorbed solution in grams = (T3 − T1);
C = mass (g) of preservative in 100 g of solution;
V = Cube volume in cm3.

2.4. Test Fungus Preparation

A strain from Trametes versicolor (L. ex. Fr.) Pilat was utilized as the biodegrading agent. The strain (CFNL 01760) was donated by the Faculty of Forestal Sciences from the Autonomous University of Nuevo Leon (Monterrey, Mexico). The strain (kept in agar test-tubes at 5 °C) was maintained viable by performing two reinoculations. The mycelium for the bioassay tests was obtained from the two-time reinoculated Petri dishes. The inoculated Petri dishes were incubated at 28 °C.

2.5. Bioassay

The fungitoxic activity of extracts was evaluated using the soil/block test methodology (AWPA E10-22) [60], which is based on determining the weight loss of the wood after a fungal attack.

2.5.1. Soil Analysis

Analysis was performed on soil samples free of fumigation chemicals. First, the soil was dried and sifted. The particles that passed through the 40-mesh sieve were selected for further preparation. The soil particles were dried at 104 °C for 24 h and later placed in a desiccator to cool down to room temperature. Then, 90 mL of soil was weighted.
The pH of the soil was determined as follows: 10 g of soil was placed in 50 mL of distilled water and shook for 20 min. The pH of the slurry was measured in a pH meter equipped with a calomel electrode (Ag/AgCl).
In order to calculate the water retention capacity, 50 g of soil was placed in a beaker, and water was poured on the soil until it was just submerged. The slurry was left for 12 h. After humectation, excess water was removed using vacuum in a Büchner flask for 15 min. Later on, 5 humected samples were weighted (Ww) and then dried at 105 °C for 24 h. The weight of dried samples was also recorded (as Wd). Retention capacity was calculated using Equation (3).
Water retention capacity (%) = (Ww − Wd)/Wd × 100

2.5.2. Culture Bottles

Glass flasks (450 mL of capacity) with plastic screw caps were used as culture bottles. According to the procedure stated in the AWPA E10-22 standard [60], water (47 mL) and soil samples (118 g) were placed in each chamber. The soil samples had a pH 6.12, 131 g of normalized volume, and a 32.37% water retention capacity.
Inoculation was performed in a laminar flow hood in sterile conditions. A feeding block was added for fungus propagation, and the flasks were prepared with caps loosened 1/4 turn. Sterilization was carried out in an autoclave for 30 min at 121 °C and 15 lb/in2 of pressure. Once room temperature was reached, inoculation of feeding blocks was completed in aseptic conditions. Blocks were inoculated with the previously prepared mycelium, except the blanks. The inoculum, 1 cm2, was obtained from zones around the colonies using sterilized punch tools, approximately from the same radial distance. The inoculum were then placed on the middle of the blocks, which were incubated for 3 weeks to ensure fungal development.
The cubes were sterilized in dried conditions for 20 min. Once room temperature was reached, the cubes were placed in culture bottles and incubated for 12 weeks, in the absence of light, at 27 °C, and relative humidity of 70%.
Once the test was finished, the cubes were cleaned (mycelium removal) and weighted (Tm weight). Then, the cubes were dried at 60 °C until they reached constant weight (recorded as T4). In order to determine the weight loss of the cubes exposed to fungi, the following equation was used:
% WL = (T3 − T4)/T3 × 100
where
WL = Weight loss;
T3 = Initial weight of the cube after impregnation;
T4 = Final weight of the cube after fungal exposition.

2.6. Leaching

The leaching material was evaluated according to the AWPA E11-16 (2006) standard [61]. Table 1 shows the quantity of impregnated cubes utilized for each formulation. The specimens were impregnated with bi-distilled water using a vacuum of 39 kPa for 30 min. Then, the cubes were submerged in water for 2 weeks, with the water containing leachate being changed with fresh water every 24 h. The water was discarded. After that, samples were dried at 60 °C until reaching constant weight (Tlix). Finally, weight loss was calculated according to Equation (5) with an alteration—by changing T4 for Tlix.
WLlix (%) = (T3 − Tlix)/T3 × 100
where
WLlix = Weight loss by leaching;
T3 = Initial weight of cube after impregnation;
Tlix = Fina weight of cube after leaching.

2.7. Statistical Analysis

All samples were analyzed according to the guidelines of the AWPA E-10 [60] and AWPA E-11 [61] standards, and results were reported as statistical means. Statistical analysis (two-way ANOVA) was carried out using OriginLab. The two factors, A and B, were the lignocellulosic starting materials (agave leaves, corn cob, and oak bark) and the concentrations of extracts were 0.1, 0.5, 1.0, 3.0, and 5.0 g/L, respectively. The p-values were reported as pA, pB, and pAB to determine if A and B are significantly different and if their interaction is significant. The significance level was set at 0.05.

3. Results

3.1. Extractions

Extracts from the leaves of agave had a dark brown color and an oily appearance, while those from the corn cob had a light brown color, with a pasty consistency. The bark extracts had a black color with a slightly oily appearance (Figure 3). Extraction yields for the agave leaves, corn cob, and bark were 9.4%, 12.4%, and 13.6%, respectively.

3.2. Soil Analysis

Results of the soil analysis of the three different samples, all taken from the property where the Center of Exact Sciences and Engineering (University of Guadalajara) is located, are shown in Table 2.
The standard values for the pH of the soil must be between 5 and 8, the water retention capacity must be in the range of 20 and 40%, and the normalized volume regarding the 118 cc weight must be higher than 90 g. Samples 2 and 3 are within these requirements; however, sample 3 had a higher water retention capacity, which helps to retain the necessary humidity inside the culture bottles during testing time. For this reason, soil sample 3 was chosen.

3.3. Impregnation of Preservative

Figure 4 shows the absorptions results from Aile wood samples impregnated with biocide substances. A slight increment in the absorption was observed when the concentration of extracts increased in the impregnation solution. Absorption average values where very similar, between 646 and 672 kg/m3. These values are within the range of high absorption according to Guevara-Salnicov (1996) [59]. The statistical analysis showed that pA > 0.0 and pB > 0.05, which indicated that, independently of the origin and concentration of the extracts, the absorption was very similar in all evaluated samples.
Average values of the retention of biocide substances and retention capacities as a function of biocide concentrations are shown in Table 3 and Figure 5, respectively. The retention increased when the biocide concentration increased. A linear relationship between retention and the concentration of biocide solution was observed. This behavior was also observed by Ramirez et al., (2012) [62]. A linear regression analysis was performed using ORIGIN 8.0 software. Extract retentions were not significantly different in relation to the lignocellulosic starting material (pA > 0.05). On the other hand, extract retentions were significantly different with regard to the concentration of used extract solutions (pB < 0.05). Also, the interaction between the lignocellulosic source and extract concentration was significant (pAB < 0.05).

3.4. Rooting Evaluation

Figure 6 shows the fungal development during the test. The inoculated feeder block (Figure 6a) was totally covered by the fungus in 3 weeks (Figure 6b). The cubes were exposed to the fungus (Figure 6c) and covered by the fungus 6 weeks after the test started (Figure 6d). Figure 6e shows the totally covered cubes. Guzman et al. (2010) reported that Aile wood is highly susceptible to fungal attacks (class V according to DIN WN 350-2) [63].
The values of the average weight loss of impregnated blocks are displayed in Table 4. The two-way ANOVA analysis showed that the weight loss was significantly different between the lignocellulosic sources (pA < 0.05) of extracts and between concentrations of biocide solutions (p < 0.05). The analysis also showed that the interaction between the source of extracts and the concentration of solutions was significant.
The weight loss as a function of the retained biocide substance’s quantity, in the wood cubes, is shown in Figure 7.
Bark extracts show a promising potential as wood preservatives. Figure 5a shows that retention increased when the concentration of bark extracts was higher in the impregnation solution. On the other hand, the weight loss decreased when the quantity of retained bark extract increased (Figure 7a), which was consistent with the lesser fungal growth and lesser mechanical damage on the impregnated cubes. Moreover, decay resistance was notably improved when using a 1.0 g/L concentration. Based on the decay resistance classification that appears in the ASTM D 2017-05 standard, 1.0 g/L solution-impregnated cubes can be classified as resistant (19% of average weight loss). In terms of average weight loss percentages, the standard states the following wood decay resistance classification: highly resistant (0%–10%), resistant (11%–24%), moderately resistant (25%–44%), and nonresistant/slightly resistant (45% or more) [64].
In the case of corn cob extracts (Figure 7b), the weight loss decreased to approximately 22% at a retention of 0.067 kg/m3, while it decreased to 25% when the retention was 0.677 kg/m3. At 2.028 kg/m3, the weight loss reduction was 35% compared to the blank blocks. The best result in terms of decay resistance is the average weight loss obtained from 3.0 g/L solution-impregnated cubes (40.69%), which can merit a moderately resistant classification according to the ASTM D 2017-05 standard [64].
For the extracts of agave leaves (Figure 7c), there was no statistically significant difference between weight loss values for the 0.1 g/L and 1.0 g/L concentrations (Table 4). However, both promoted fungal inhibition in the wood since the average weight loss decreased by 25% in comparison to blocks without extracts. Blocks impregnated with a 5.0 g/L solution had a 40% reduction and an average weight loss of 32.99%, which is the lowest obtained for this type of extract. In the same line of analysis of the rest of extracts, based on the classification that appears in the ASTM D 2017-05 standard [64], the decay resistance of 5.0 g/L solution-impregnated wood cubes is moderate. In general, a higher content of retained extracts leads to a lower weight loss.

3.5. Leaching

The retention of biocides in impregnated products is not only important to achieve resistance against fungi in the wood, but it is also relevant regarding their impact on the environment and human health due to their release to air, water, and soil. Leaching (at 14 days) of extract-impregnated Aile blocks is shown in Figure 8. It was observed that when retention of the biocide substance increased, the quantity of leachate also increased. The extracts that presented higher leaching were those of agave, followed by those from corn cob and oak bark, in this order. ANOVA showed that leaching was not significantly different between lignocellulosic starting materials (pA > 0.05), as well as for extract retention (pB > 0.05). The interaction between the lignocellulosic origin and retention is not significant.

4. Discussion

The extracts, also known as secondary metabolites, are mainly fat, fatty acids, fatty alcohols, phenols, terpenes, steroids, resin acids, rosins, waxes, quinones, amino acids, phenylpropanes, lignans, flavonoids, inorganic substances, and many other organic compounds [65]. All those components exist as monomers, dimers, and low-molecular-weight polymers. Previous studies showed that the most successful mixture to obtain extracts is the acetone/hexane/water (54:44:2) mixture [7,66,67,68,69], from which essential oils, resins, tannins, and phlobaphenes are regularly extracted.
Our results in the case of extract production (yields of extracts), using the hexane/acetone/water solvent system, are in the range of those reported in similar studies. García (2000) reported extractions from the bark of several conifers, obtaining yields between 8.89 and 12.8% [7]. In another study, Torres (2004) performed extractions with ethanol on corn cobs and agave leaves, obtaining yields of 6.35 and 7.19%, respectively [70]. Velásquez et al. (2006) reported yields of 11.0, 15.24, and 13.48% using acetone, ethanol, and water, respectively, for carob heartwood’s extracts. Moreover, in the same study, yields of 11.0, 16.57, and 8.68% were obtained for Puy heartwood´s extracts using the same solvents [71]. Ramírez et al. (2012) reported a 13.5% yield of extracts obtained from the mesocarp of Cocus nucifera Liem utilizing a 2% sodium bisulfate solution [62]. It is evident that differences of extract yields in the literature could be attributed to many factors, such as the studied species, part of the tree/plant evaluated, solvents used, hydromodule system used, etc.
It is well known that preservative products will not provide the desired effect if they are not properly used (OIMT, 1989). The ideal scenario is to impregnate wood with a highly effective product, resistant to leaching losses or evaporation, and in sufficient doses to achieve a homogeneous distribution of active substances. When wood is treated, its properties, the nature of the preservative, and its way of usage must be considered. Ideal protection can be obtained if these factors are working synergistically. The method to incorporate the preservative should be one that allows deep penetration of the preservative within the wood. The permeability of wood is a property that approximately indicates the ability of a substance to penetrate it. This permeability will determine the quality of the impregnation, which varies according to the methods employed and to the involved penetration phenomena [72].
The cubes were impregnated using the full-cell method via a vacuum–pressure–vacuum cycle. A common belief is that the purpose of the initial vacuum is to remove the water within the wood, which is impossible considering the pressure, temperature, and duration of these processes. The initial vacuum seeks to remove the air from the cylinder and cellular lumens, which facilitates the incorporation of the biocide solution to the wood. In the present study, five cubes were cut longitudinally, confirming that the biocide solution penetrated to the center of the cubes. Machuca-Velasco et al. (2005) impregnated wood from Spondias mombís L. using copper, chrome, and arsenic (CCA) salts, pentachlorophenol, and creosote, reporting absorption values of 560.75, 22.95, and 174.55 kg/m3, respectively [73]. Also, Otaño et al. (1999) reported absorption values of 583.71, 571.64, 560.54, and 477.3 kg/m3 for woods from Pinus halepensis Mill., P. pinaster Ait., P. pineal. L., and P. radiata D. Don., respectively [74]. In this work, absorption results were classified as high absorption (HA) and were in the range of results reported for low-density woods. Moreover, absorption values were practically the same for all evaluated samples, which indicates high reproducibility and reliability of the utilized impregnation process.
Lomelí et al. (2012) reported retention values from 2.023 to 14.03 kg/m3 using extracts from Aile wood [69]. These values are very high, which suggests that such extracts could not be used as low-cost natural preservatives. In another study, Aburto (2006) reported the impregnation of wood from Mangifera indica L. using boron salts, obtaining a value of 7.5 kg/m3 [75]. Moreover, Díaz (2014) obtained a lower retention value (4.1 kg/m3) in Q. rugosa´s wood [76]. In our case, retention values were lower, mainly due to the concentrations of biocide solutions (≤5.0 g/L).
Several studies have reported the presence of biocide substances in bark extracts. Hathway (1958) reported the presence of catechin, gallocatechin, leucodelphinidin, and phlobatannins in the bark of Quercus pedunculata Ehrh and Quercus sessiliflora Salisb [77]. In another study, Parker (1977) reported the presence of catechins, quercitrin, robinin, quercetin 3-methyl ether, scopoletin, chlorogenic acid, various leucoanthocyanins, and condensed and hydrolyzable tannins in the bark of Quercus velutina Lamarck (black oak) [78]. Stirling et al. (2007) evaluated the individual components of extracts from the wood of occidental red cedar and determined their capacities as biocides, metal chelators, and radical removers. They found that the thujaplicins, β -thujaplicinol, and the plicatic acid were toxic to wood-decay fungi and were good metal chelators [79]. Extracts from the bark of Acacia mollissima and from the heartwood of Schinopsis lorentzii showed fungal resistance, while extracts from the bark of Pinus brutia were ineffective when tested against white- and brown-rot fungi. In the case of extracts from agave leaves, Torres et al. (2004) reported that ethanolic extracts from the leaves of agave inhibited the mycelial growth of Trametes versicolor [70]. Morales-Serna et al. (2010) studied the homoisoflavanones of the leaves of Agave tequilana Weber, in which they identified sitosterol and stig-masterol as the main components of the hexane extract, while glucose, sucrose, and fructose were obtained from the methanolic extract. From the acetone extract, homoisoflavanones: 5,7-dihydroxy-3-(4-methoxybenzyl)-chroman-4-one (1), 7-hydroxy-3-(4-hydroxybenzyl)-chroman-4-one, and 4′-demethyl-3,9-dihydro-punctatin were able to be isolated [79]. Studies dealing with corn cob extracts were not found.
When the retention values of the extracts in wood samples increased, the weight loss of all analyzed species decreased [80]. In this work, we observed that by evaluating inhibitory effect in terms of weight loss, good results were obtained. For example, bark extract-impregnated cubes could be classified as resistant according to the ASTM D 2017-05 standard [64]. Similarly, cubes impregnated with agave leaves and corn cob both could be classified as moderately resistant. Bark extracts provided the best protection, which could be due to the protective role of the bark. We also believe that better protection against fungal attacks can be achieved if the concentration of the extract increases. However, based on the results of the present study, one can argue that using corn cob and agave leave extracts could be too expensive due to the high amount of extracts needed to achieve wood resistance.
Goktas et al. (2007) reported weight losses between 8.64 and 24.06%, caused by Trametes versicolor in non-impregnated wood, and from 5.02 to 28.25% with wood impregnated with extracts of non-timber products such as Nerium oleander [81]. Also, Ramirez et al. (2012) reported weight losses of 71.04, 61.62, 57.71, and 57.09% for extracts from the mesocarp of Cocos nucifera linn using impregnation solutions of 0.5, 1.0, 2.0, and 4.0 g/L, respectively. The blank sample displayed a 73.0% weight loss [62].
Regarding the blank sample, we can say that the weight loss values for Aile wood without impregnation were consistent with the weight losses of other woods in the same period of time and by the same fungus, T. versicolor [62]. In the case of the biocidal substances evaluated in this work, it was observed that all of them inhibited T. versicolor. The most encouraging extracts are those obtained from the bark of Q. laeta, followed by those from the corn cob and agave leaves, in this order.
Given that water exposure of treated wood represents the main route for the emission of toxic biocides into the environment, it is necessary to study their mechanisms and methodologies to evaluate the toxic effects exhibited by leaching processes. The degree of biocide release from treated wood in contact with water is a crucial parameter that determines the magnitude of negative effects on human health and other living organims. Corn cob and bark extract displayed less leaching than in comparison to agave leaves, probably because they had a better affinity with Aile wood. The release of biocides from treated wood products is the result of coupled chemical reactions, transport processes, and biological activity. In contact with water, the wood material undergoes chemical and structural changes, releasing soluble compounds, such as natural mineral salts and extractives. Fixed biocides are partially desorbed after achieving a thermodynamic equilibrium [82].
Little information has been published on the mechanism of organic biocides’ fixation in wood materials [83]. However, it was observed that these molecules are resistant to leaching, suggesting that they undergo possible interactions with wood structures. It is expected that organic biocides interact with different components of wood, at least through weak physical bonds (van der Waals, dipolar, and charge transfer interactions), thus adsorbing biopolymers. The distribution of biocides between the aqueous solution and wood polymers depends on their hydrophobicity and aqueous solubility. Therefore, hydrophobic molecules are expected to have more affinity for lignin than for cellulose. Stronger interactions can occur when biocides have functional groups capable of reacting with specific sites in the wood, such as hydroxyls (hydrogen bonds). In this context, it is reported that the leaching of biocidal substances from impregnated wood is significantly affected by the preservative’s concentration, the conservation method, and wood species [84]. Also, it is possible to improve the fixation of extracts by introducing other substances. For example, Sen et al. (2009) reported the use of boron salts to improve the extract fixation of Quercus macrolepis Ky, the bark of Pinus brutia, and the leaf powder of Rhus coriariae [8].

5. Conclusions

In this work, the soil block method was used to evaluate the degree of biodeterioration caused by T. versicolor in Aile wood, which was impregnated with extracts obtained from the bark of Q. laeta, the cob of Zea mays, and the leaves of A. Tequilana. The bark of Q. laeta had the highest extraction yield (13.6%) and, in the case of the impregnation procedure, there were no significant differences in average absorption values between concentration groups and different types of extracts. Moreover, a linear relationship was observed between the retention and the concentration of biocide solutions. The wood cubes that had the lowest average weight loss (19.38%) were the ones impregnated with extracts from the bark of Q. laeta, which can greatly improve the decay resistance of Aile wood from being nonresistant to resistant, according to the standard ASTM D 2017-05. In general, a higher content of retained extracts leads to a lower weight loss. It was also observed that, by increasing the retention of the biocide substance, the amount of leachate also increased. The extracts that showed the greatest leaching were those from agave, followed by those of corn cob and oak bark. Based on these results, it is clear that the most promising type of extract with a potential for being used as a natural preservative for woods is the one from the bark of Q. laeta. Therefore, it is important to focus future works on the identification of the bioactive substances responsible for its fungitoxic capacity.

Author Contributions

Conceptualization, M.G.L.-R., J.G.T.-R. and S.G.-E.; methodology, A.G.-M., J.A.-H., M.G.L.-R. and S.G.-E.; validation, A.G.-M., J.G.T.-R. and R.M.J.-A.; formal analysis, A.G.-M., J.A.-H., M.G.L.-R. and S.G.-E.; investigation, J.G.T.-R., J.A.S.-G., M.G.L.-R. and S.G.-E.; resources, J.A.S.-G. and S.G.-E.; data curation, J.G.T.-R. and S.G.-E.; writing—original draft preparation, R.M.J.-A., J.G.T.-R. and S.G.-E.; writing—review and editing, R.M.J.-A., J.G.T.-R. and S.G.-E.; supervision, J.A.S.-G. and S.G.-E.; project administration, J.G.T.-R. and S.G.-E.; funding acquisition, M.G.L.-R. and S.G.-E. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Data Availability Statement

Data are contained within the article.

Conflicts of Interest

The authors declare no conflicts of interest.

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Figure 1. Diagram of the methodology.
Figure 1. Diagram of the methodology.
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Figure 2. (a) Representation of the impregnation equipment. (b) Vacuum–pressure cycle used in the impregnation process.
Figure 2. (a) Representation of the impregnation equipment. (b) Vacuum–pressure cycle used in the impregnation process.
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Figure 3. Extracts from (a) agave leaves, (b) corn cob, and (c) oak bark.
Figure 3. Extracts from (a) agave leaves, (b) corn cob, and (c) oak bark.
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Figure 4. Absorption of different concentrations of biocide solutions used to impregnate Aile wood samples.
Figure 4. Absorption of different concentrations of biocide solutions used to impregnate Aile wood samples.
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Figure 5. Extract retention as a function of the concentration of biocide solution. Every colored shape represents a single evaluated sample.
Figure 5. Extract retention as a function of the concentration of biocide solution. Every colored shape represents a single evaluated sample.
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Figure 6. (a) Inoculation in culture bottles; (b) fungal growth in the feeding block; (c) cubes exposition to the fungus; (d) culture bottles at 6 weeks and (e) at test completion.
Figure 6. (a) Inoculation in culture bottles; (b) fungal growth in the feeding block; (c) cubes exposition to the fungus; (d) culture bottles at 6 weeks and (e) at test completion.
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Figure 7. Weight loss as a function of the quantity of retained biocide substance. Every colored shape represents a single evaluated sample.
Figure 7. Weight loss as a function of the quantity of retained biocide substance. Every colored shape represents a single evaluated sample.
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Forests 15 01204 g008
Figure 8. Average leaching percentage as a function of the average retention of biocide substances (14th day).
Figure 8. Average leaching percentage as a function of the average retention of biocide substances (14th day).
Forests 15 01204 g008
Table 1. Numbers of cubes impregnated for leaching and bioassay tests.
Table 1. Numbers of cubes impregnated for leaching and bioassay tests.
Concentration (g/L)Number of Cubes Impregnated with Extracts
Agave LeavesCorn CobOak Bark
LeachingBioassayLeachingBioassayLeachingBioassay
0.0101610161016
0.1101610161016
0.5----1016
1.0101610161016
3.0--1016--
5.01016----
Table 2. Analysis of soil samples.
Table 2. Analysis of soil samples.
SamplespHWater Retention Capacity (%)Normalized Volume (g)
14.86 ± 0.0929.18 ± 1.89114.9 ± 3.12
25.73 ± 0.0822.61 ± 1.26108.4 ± 2.73
36.12 ± 0.0932.37 ± 1.72131.0 ± 3.26
Table 3. Average retention of biocide substances in Aile samples.
Table 3. Average retention of biocide substances in Aile samples.
Concentration (g/L)Average Retention (kg/m3)
Agave LeavesCorn CobBark Oak
0.00.0000.0000.000
0.10.065 ± 0.0060.067 ± 0.0670.067 ± 0.007
0.5--0.338 ± 0.026
1.00.653 ± 0.0430.677 ± 0.0770.621 ± 0.051
3.0-2.028 ± 0.179-
5.03.367 ± 0.218--
Linear analysis
Retention (kg/m3) = A + B × Concentration (g/L)
A0.0275 ± 0.02950.00289 ± 0.00340.0059 ± 0.0094
B0.7382 ± 0.01160.68495 ± 0.00220.6321 ± 0.0168
R20.9990.9990.998
Table 4. Weight loss of Aile wood blocks at the end of bioassay.
Table 4. Weight loss of Aile wood blocks at the end of bioassay.
Concentration (g/L)Weight Loss (%)
Agave LeavesCorn CobBark Oak
0.075.68 ± 2.475.68 ± 2.475.68 ± 2.4
0.149.77 ± 3.253.64 ± 2.544.19 ± 3.2
0.5--28.05 ± 1.9
1.047.07± 4.150.37 ± 1.9719.38 ± 2.1
3.0-40.69 ± 2.18-
5.032.99 ± 3.9--
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MDPI and ACS Style

Gálvez-Martínez, A.; Jiménez-Amezcua, R.M.; Anzaldo-Hernández, J.; Lomelí-Ramírez, M.G.; Silva-Guzmán, J.A.; Torres-Rendón, J.G.; García-Enriquez, S. Evaluation of the Fungitoxic Effect of Extracts from the Bark of Quercus laeta Liebm, the Cob of Zea mays and the Leaves of Agave tequilana Weber Blue Variety against Trametes versicolor L. Ex Fr. Forests 2024, 15, 1204. https://doi.org/10.3390/f15071204

AMA Style

Gálvez-Martínez A, Jiménez-Amezcua RM, Anzaldo-Hernández J, Lomelí-Ramírez MG, Silva-Guzmán JA, Torres-Rendón JG, García-Enriquez S. Evaluation of the Fungitoxic Effect of Extracts from the Bark of Quercus laeta Liebm, the Cob of Zea mays and the Leaves of Agave tequilana Weber Blue Variety against Trametes versicolor L. Ex Fr. Forests. 2024; 15(7):1204. https://doi.org/10.3390/f15071204

Chicago/Turabian Style

Gálvez-Martínez, Alberto, Rosa María Jiménez-Amezcua, José Anzaldo-Hernández, María Guadalupe Lomelí-Ramírez, José Antonio Silva-Guzmán, José Guillermo Torres-Rendón, and Salvador García-Enriquez. 2024. "Evaluation of the Fungitoxic Effect of Extracts from the Bark of Quercus laeta Liebm, the Cob of Zea mays and the Leaves of Agave tequilana Weber Blue Variety against Trametes versicolor L. Ex Fr" Forests 15, no. 7: 1204. https://doi.org/10.3390/f15071204

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