Revista Mexicana Ciencias Agrícolas   volume 14   number 3   April 01 - May 15, 2023

DOI: https://doi.org/10.29312/remexca.v14i3.3081

Article

Growth of Vicia faba L. seedlings in mixtures of soil with biosolids

Maribel Quezada-Cruz1

Porfirio Raúl Galicia-García1§

Serafín Cruz-Izquierdo2 

1Environmental Biotechnology Laboratory-Technological University of Tecámac. Federal Highway Mexico-Pachuca km 37.5, Col. Sierra Hermosa, Tecámac, State of Mexico, Mexico. CP. 55740. Tel. 55 59388400, ext. 452. (maryquezada9@gmail.com).

2Postgraduate in Genetic Resources and Productivity-Genetics. Postgraduate College-Campus Montecillo. Mexico-Texcoco highway km 36.5, Montecillo, Texcoco, State of Mexico, Mexico. CP. 56230. Tel. 55 59388400, ext. 452. (pgaliciag@uttecamac.edu.mx; prgaliciag61@gmail.com).

 §Corresponding author: prgaliciag61@gmail.com.

Abstract

In order to investigate the effect of four biosolids from the dairy, malt, soap and paper industries on the growth and production of biomass of Vicia faba L. seedlings, biosolid:soil mixtures were made in ratios 20:80, 40:60 and 60:40. Physicochemical and microbiological analyses were performed on each biosolid and mixtures. Biosolids showed differences in the presence (MPN) of fecal coliforms from 3 to 1 100, Salmonella sp., from 2 to 3 and from 0 to 16 helminth eggs per g of total solids. The concentration (mg kg-1) of heavy metals in copper was from 0.7 to 1.9, chromium from 0.8 to 2.4, cadmium from 0 to 0.6, zinc from 4.3 to 8.6, nickel from 1.2 to 3.8 and lead from 1.3 to 5.7, without arsenic and mercury. The contaminants were below the permissible limits according to SEMARNAT (2002), in addition, among biosolids the pH varied from 7 to 9.8 and organic material (OM) from 0.3 to 6.2%. Biosolids from the dairy and malt industries incorporated into the soil modified the pH from 8.1 to 7.4, increased OM from 1.2 to 3.5%, the total nitrogen from 7 to 35 mg kg-1 and the available phosphorus from 5 to 25 mg kg-1. At 30 days after sowing in the greenhouse, V. faba seedlings grown in soil with the biosolid from the dairy industry in the highest ratio increased biomass production by 1 145% and length by 342%.

Keywords: Vicia faba L., heavy metals, biosolids, microbiology.

Reception date: January 2023

Acceptance date: March 2023

Introduction

Urban and industrial wastewater treatment plants (WWTPs) generate sludges with a high content of organic matter and nutrients of interest to improve the soil in agricultural production systems (SEMARNAT, 2002). However, the use of industrial sludges is restricted, due to the presence of trace elements, pathogenic organisms, organic xenobiotic substances, endocrine disruptors and hormones (Bücker-Neto et al., 2017), which can induce diverse and adverse damage to plants of agricultural interest (Corrêa-Martins et al., 2016).

The chemical composition and physical properties of the sludges of WWTPs vary depending on the industry from which they come, and the fertilizers derived from such sludges as well, so before using a residual sludge, they must be stabilized to eliminate pathogens, toxic chemical compounds and unpleasant odors. This stabilization is carried out by physical, chemical and biological processes, using heat (Lang and Smith, 2008), alkalinity, aerobic or anaerobic digestion (Goss et al., 2013), among others. After being subjected to the stabilization process, such sludges are called biosolids.

Currently, soil fertility and crop productivity have decreased significantly, and chemical fertilizers are used indiscriminately (Postisek-Talavera et al., 2010). Due to the high cost of chemical fertilizer sources and the decrease in raw material, organic waste recycling has been chosen. For this purpose, biosolids are an excellent source to consider. In Mexico, more than 640 million tons of biosolids on a dry basis are produced annually (Mantilla et al., 2017), which generates serious problems for their elimination and confinement. Biosolids contain a large amount of essential nutrients for plants, so it would be convenient to use them in an agricultural production system and at the same time reduce the negative impact on the environment with a sustainable production system (Gomiero et al., 2011).

Biosolids have great potential for use as organic fertilizer, but the nutrient imbalance needs to be overcome (Antille et al., 2013), as they contain a high concentration of micronutrients and some macronutrients are not present in necessary concentrations as required by crops (Dad et al., 2018). Nutrient release rates are difficult to predict so the production response is highly variable, which limits their management from the agronomic and environmental perspective (Deeks et al., 2013).

Broad bean (Vicia faba L.) is a legume and an important source of protein for the population around the world, its consumption is in fresh and dried form by humans and animals (Vela et al., 2018), it is an excellent fixer of nitrogen in the soil, due to the characteristics of the rhizosphere (Harris-Valle et al., 2019), it is of rapid growth of 150 days and has high capacity to accumulate biomass up to 15 t ha-1 of pod (ICAMEX, 2023).

In addition, this species has genes tolerant to osmotic (Ghouili et al., 2021) and saline stress (Rajhi et al., 2020), is capable of accumulating and transferring heavy metals, such as cadmium (Piršelová et al., 2021), arsenic (Gupta et al., 2020), boron, lead, zinc, and copper (Alaoui et al., 2019), to its inedible organs such as leaves and stems, in addition, with low accumulation in edible organs such as seeds, so it is considered an interesting biological model in soil remediation together with food production, which is a necessary practice in the face of food needs in the population (Tang et al., 2019).

V. faba is important in Mexico because it is the second source of vegetable protein nationwide and in the State of Mexico the grain yield is 1.9 t ha-1, lower than the national average of 2.5 t ha-1 (SAGARPA, 2022). Therefore, it is necessary to improve its yield with sustainable fertilization alternatives. The objectives of this work were the partial characterization of biosolids from the dairy, malt, soap and paper industries, determination of their effect in different ratios of biosolid:soil in the growth and production of biomass of Vicia faba seedlings.

Materials and methods

Biosolids

Samples of each biosolid from the dairy, malt, soap and paper industries were collected in mounds of approximately 6 m3, deposited the previous day in a plot of agricultural use for incorporation into the soil. The plot is located in the municipality of Santa Ana Nextlalpan, State of Mexico (19° 40’ 50” north latitude, 99° 07’ 56” west longitude), with 2 235 m altitude. The average annual rainfall of the region is 600 mm distributed from April to September with more than 80% and a dry season from October to March, the average annual temperature is 14.1 °C with a minimum of 2.3 and a maximum of 31 °C, the soil is of lacustrine origin with predominantly alkaline pH, because it is located in the vicinity of what was the former lake of Xaltocan (Velasco, 2010).

Before their incorporation into the soil, by means of the quartering method, a sample composed of 15 kg of each biosolid, which was deposited in polyethylene bags, and a sample of 10 g of each one in sterile containers for microbiological analyses, were obtained from each mound. Each biosolid underwent a microbiological analysis that consisted of quantifying the most probable number (MPN) of fecal coliforms, Salmonella spp., and determining helminth eggs. Soil samples without biosolid (control) were collected in a plot 300 m away from the plot with biosolids.

The soil and biosolid samples were subjected to digestion with nitric and sulfuric acid, to subsequently quantify the concentration (mg kg-1) on a dry basis of the heavy metals of copper (Cu), chromium (Cr), cadmium (Cd), zinc (Zn), nickel (Ni), lead (Pb), mercury (Hg) and arsenic (As), by means of an atomic absorption spectrophotometer (SavantAA, GBC Scientific Equipment, USA). The standard was a mixture of trace elements at 1000 ppm (SCP Science). All of the above was carried out in accordance with Mexican standards (SEMARNAT, 2002). In addition, the pH and percentage of organic matter (OM) were determined using the method of Walkley and Black (SEMARNAT, 2000).

Biosolid-soil mixture

Biosolid-soil mixtures weighing 15 kg of each of the four biosolids from the dairy, malt, soap and paper industries were carried out, in the ratios of 20:80, maximum ratio used by González-Flores et al. (2017) in corn (Zea mays L.); that is, 20% biosolids with 80% soil, as well as biosolid:soil mixtures 40:60 and 60:40. Also, a mixture was made with the four biosolids in the ratio 25:25:25:25 and a control sample (soil without biosolids).

Samples of 1.5 kg of each mixture and of the control were considered for laboratory analysis. In three repetitions of each mixture and the control, the pH was determined by means of a potentiometer (Hanna pH112), bulk density (g cm-3), content (mg kg-1) of nitrogen (micro-Kjeldahl) and available phosphorus (Olsen) on a dry basis, in addition, the percentage of organic matter (Walkley and Black) in each of the mixtures according to SEMARNAT (2000) for soil fertility analysis.

Experiment in the greenhouse

Fourteen substrates were used, which resulted from mixing in three different ratios each of the four biosolids with soil, the mixture in equal parts of the biosolids and the control substrate. For the experiment, seeds of Vicia faba var. ICAMEX-31 were used, the seeds were disinfected with a 0.6% sodium hypochlorite solution for 2 min and rinsed with distilled water several times, a pre-seed germination analysis was performed on the substrates to ensure the number of repetitions, the percentage of germination decreased in some biosolid-soil mixtures from 100 to 90%; nevertheless, at least one seedling was obtained on each substrate and in each of the four repetitions.

The experimental unit consisted of a pot 15 cm high and 20 cm in diameter containing 2 kg of substrate, three seeds were sown in each pot 10 cm away and 3 cm deep in the substrate, four repetitions per treatment were performed. The pots were placed in the greenhouse and humidity was maintained at field capacity at a temperature between 18 and 28 °C during the test period.

At 30 days after sowing, the whole seedlings were harvested, the roots were separated from the aerial part (leaves and stem) and the length (cm) of each section was measured, then they were added to obtain the total length of the seedlings. Subsequently, the stem and root of each plant were placed separately in an oven (Ríos Rocha®) at 80 °C for 3 days until they reached a constant weight, and the dry weight (g) was determined on an analytical balance. The total biomass of each seedling was obtained by adding the dry weights of the stem with leaves and root.

Statistical analysis

The experiment was established with a completely randomized design with a factorial arrangement; with the factor of biosolid:soil mixtures with four levels (dairy:soil, malt:soil, soap:soil and paper:soil), with the factor of biosolid:soil ratios in three levels (20:80, 40:60 and 60:40); the mixture of the four biosolids and soil. The data were subjected to analysis of variance, when statistically significant differences were found, the means were separated by the Duncan test (p≤ 0.05). In all cases, the software of the statistical analysis system (SAS) v.6.12 was used.

Results and discussion

Biosolid analysis

Fecal coliforms and Salmonella spp. were found in all biosolids. With regard to helminth eggs, the only biosolid that presented these parasites was that of the dairy industry. All biosolids showed values below the maximum permissible limits. Those from the malt, soap and paper industries were in class A (Table 1), are suitable for application in urban properties. That from the dairy industry had a class C profile, recommended for use only in forest soils without contact with people due to the number of pathogens present (SEMARNAT, 2002).

Table 1. Pathogens, parasites and classification of biosolids from the malt, soap, paper and dairy industries.

MPN= most probable number; TS= total solids. A= for urban use in contact with people during its application; C= forest use, only improvement without contact with people (SEMARNAT, 2002).

In order to characterize biosolids for agricultural use, organic matter (OM) and pH were determined according to Mexican standards (SEMARNAT, 2000). The biosolids from the dairy, malt, paper and soap industries, their OM contents were, respectively, 6.2%, 2.5%, 1.5% and soap 0.4%. The pH of biosolids from the dairy and paper industries was 7 and 7.3, respectively, that of the malt industry was 7.6, and the biosolid from the soap industry was 9.8 (Table 2).

Table 2. Organic matter (OM) and potential of hydrogen (pH) of biosolids from dairy, malt, soap and paper industries.

pH= strongly alkaline = +; moderately alkaline = ±; organic matter (OM), very low = ♦; low = ♦♦; medium = ♦♦♦; very high = ♦♦♦♦♦ (SEMARNAT, 2002).

These results show that biosolids from the malt and paper industries can be useful to improve soil characteristics, due to low concentrations of pathogens (fecal coliforms and Salmonella spp.) and they do not present parasites (helminth eggs). They also show a good percentage of OM and a pH suitable for plant growth. However, the biosolid from the dairy industry, even with good characteristics of OM and pH, needs a more rigorous stabilization process to remove parasites. In relation to the biosolid from the soap-manufacturing industry, its usefulness in agriculture should be reconsidered because Price et al. (2021) report that due to its characteristics, it has an effect on the decrease of beneficial soil microfauna.

The four biosolids included essential elements for plants such as nickel, copper and zinc, others such as chromium, cadmium and lead that are not necessary, in no case mercury and arsenic were detected. Copper was only found in biosolids from the dairy and soap industries (Table 3). All concentrations were below the maximum permissible limits indicated by the Mexican standard (SEMARNAT, 2002) and therefore useful to add them to the soil for agricultural purposes.

Table 3. Concentration of heavy metals in biosolids from the dairy, malt, soap and paper industries.

Below the limit of detection (BLD). Limit of detection of (SavantAA, GBC Scientific) for Cu (0.05), Cr (0.17), Cd (0.009), Zn (0.4), Ni (0.04), Pb (0.06), Hg (1.6), As (0.04). n = 3; ± standard deviation.

Previous reports in Mexico of analysis of biosolids product of urban WWTPs show a concentration of heavy metals higher than those obtained in this work, although within the maximum permissible limits of the standard. In the state of Puebla, Mexico, González-Flores et al. (2017) reported 25.09 mg kg-1 copper and 163.99 mg kg-1 zinc. In Durango, Mexico, Flores-Félix et al. (2014) reported concentrations (mg kg-1) as follows: 266 copper, 14.6 chromium, 899 zinc, 11.7 nickel and 65 lead, in addition to elements such as arsenic and mercury of 12.1 and 1.23 mg kg-1, respectively.

Physicochemical analyses of biosolid mixtures

Soil, when adding the biosolid from the dairy industry to the soil, the pH decreased from 8.1 to 7.2 (Table 4). The same effect, but to a lesser degree, was observed when using malt (8.1 to 7.3) and paper (8.1 to 7.5) biosolids. The biosolid from the soap industry showed an opposite effect, increasing the pH of the soil from 8.1 to 8.8, in this case, to produce soap or detergents, the industries use hydroxides for saponification.

Table 4. Organic matter (OM), potential of hydrogen (pH), total nitrogen (N-total) and available phosphorus (P), in mixtures of biosolid with soil in different ratios and soil without biosolid (control).

(OM)= very low =♦; low =♦♦; medium =♦♦♦; high =♦♦♦♦; very high =♦♦♦♦♦. Available (P)= low =♦; medium =♦♦; high =♦♦♦; N-total= total nitrogen: very low =♦; low =♦♦; medium =♦♦♦; high =♦♦♦♦; very high =♦♦♦♦♦ (SERMARNAT, 2000). n= 3 ± standard deviation.

The soil samples had on average 1.2% OM, mixing the soil with some of the biosolids increased the OM of the mixtures, but in different amounts. Mixtures with biosolids from the dairy industry with soil had on average 5.5% OM, those from the malt industry with soil had on average 3.2% OM, those from the paper industry with soil had 1.8 OM and mixtures of biosolids from the soap industry with soil had 1.2% OM.

In the same order of mixtures, these provided nitrogen (N-total) and available phosphorus (P). On average, biosolid:soil mixtures of the dairy industry had 55 mg kg-1 of N-total and 34 mg kg-1 of available (P); those of the malt industry with soil had 26 mg kg-1 of N-total and 32 mg kg-1 of available (P), those of biosolids from the paper industry with soil had 19 mg kg-1 of N-total and 7.8 mg kg-1 of available (P), those of soap with soil had 12 mg kg-1 of N-total and 20 mg kg-1 of available (P). Finally, the mixture of the four biosolids had a pH of 6.9, 1.4% OM, 12 mg kg-1 of N-total and 28 mg kg-1 of available (P), according to the standard in Mexico (SEMARNAT, 2000).

Analysis of seedling growth and biomass

At 30 days after sowing, the analysis of variance (Table 5), revealed that the biosolid:soil mixtures of the four industries evaluated, as well as in the three different ratios, had highly significant effect differences (p≤ 0.001) on the biomass and length of V. faba seedlings, it also evidenced that there was no interaction between the mixtures and the ratios; that is, the effect of biosolids on soil was equal to the effect of ratios on both variables.

Table 5. Mean squares and statistical significance of biomass (g) and length (cm) of broad bean (Vicia faba L.) seedlings grown in biosolid:soil mixtures.

ns= non-significant difference; **= highly significant difference (p≤ 0.01).

Figure 1 shows the average biomass and length of broad bean seedlings. Biosolids from the dairy and malt industries added to the soil achieved the greatest effect on biomass production 1.7 g and 1.6 g, respectively. Biosolids from the soap and paper industries also increased biomass, although to a lesser extent, 1.4 g, compared to soil without biosolid (1.1 g).

Figure 1. Average biomass and length of broad bean (Vicia faba L.) seedlings grown in soil without biosolids (control), biosolid mixtures and biosolid:soil mixtures. Means with the same letter are statistically equal (Duncan, p≤ 0.05) in each variable.

This same effect was observed in the length of seedlings with biosolids from the dairy and malt industries, which produced an increase three times more their length 56.3 and 55.8 cm, compared to soil without biosolid 12.7 cm and the length increased with soap and paper up to 40 and 47 cm. With these results it is inferred that the biosolids from the dairy, malt, soap and paper industries individually modified the characteristics of the soil and as a consequence, a highly significant increase in the biomass and length of V. faba seedlings was detected. In the case of seedling root biomass, no significant effect was observed on biomass and its length when adding biosolids to the soil, similar to what was reported by Vela et al. (2018).

Seedlings that grew in biosolid-soil mixtures of the soap and paper industries showed lower biomass than those that grew in malt and dairy industry mixtures but higher biomass than seedlings that grew in soil without biosolid. From this observation, it follows that the low amount of organic matter and the alkaline pH decreased the adequate conditions of growth and nutrition, which limited the increase in biomass and length of the seedlings, even though V. faba has the capacity to grow in unfavorable environments such as pH and salt stress (Belachew and Stoddard, 2017).

In the biosolid:soil mixtures 20:80, the average biomass was 8.8 g, in 40:60 it was 10.4 g and in 60:40 it was 13.4 g; that is, a gradual increase in biomass was observed as a result of the increase in the amount of biosolid that was mixed with soil (Figure 2). Therefore, it follows that the increase in the amount of biosolids in the soil gradually improves soil characteristics and manifests itself in highly significant differences for the biomass and length of V. faba seedlings. This is consistent with what was reported by González-Flores et al. (2017), who obtained an increase in the height of adult corn plants from 34 to 114 cm and an increase in biomass from 9 to 125 g, by adding biosolids from an urban WWTP to the soil.

Figure 2. Accumulated biomass of broad bean (V. faba L.) seedlings, grown for 30 days from sowing in soil without biosolid, biosolid mixtures, biosolids:soil mixtures in ratios 20:80, 40:60, and 60:40. Means with the same letter are statistically equal (Duncan, p≤ 0.05).

However, in Figures 1 and 2, it is observed that the seedlings that were in the substrate that was the mixture of the four biosolids, their length was 19.4 cm which represents only 15.3% of the length in the biosolid from the dairy industry with 56.3 cm, and less accumulation of biomass of 0.7 g which represents 136.3% less than the seedlings in the mixture 80% of dairy biosolids and 20% of soil. This can be explained by the fact that it had a pH of 6.9, minimum concentration of OM and N-total and large amount of available (P), in addition to having a sticky consistency and poor drainage; that is, an inappropriate mixture so that nutrients were available for seedlings, so in the field it should be avoided.

When considering the recommendation of fertilization of V. faba, in the study area (INIFAP, 2017), the four biosolids contribute in a very variable way with the amount of nitrogen and phosphorus to cover the needs of the crop. With the exception of nitrogen provided by the biosolid from the dairy industry, the other biosolids provide less than half the nitrogen and phosphorus needed for the crop. This can be corrected if the necessary element or elements are added in a sufficient dose of biosolid to the soil (Sandaña et al., 2018).

Interestingly, our results show that the biosolids that come from the WWTPs of the studied industries located in Mexico have a concentration of heavy metals up to ten times lower (Bedoya-Urrego et al., 2013; Flores-Félix et al., 2014; González-Flores et al., 2017), minimal amount of pathogenic microorganisms (Flemming et al., 2017; Hernández et al., 2017) and do not include metals such as arsenic and mercury, important differences with the biosolids from urban WWTPs, which indicates that some biosolids of industrial origin are favorable for use in agriculture.

Conclusions

Biosolids from the dairy, malt, paper and soap industries showed differences in potential of hydrogen, percentage of organic matter, concentration of heavy metals, total nitrogen and available phosphorus, and microbiological differences. Mixing the soil with biosolids from the dairy, malt and paper industries significantly improved the previous characteristics, decreased the pH from 8.1 to 7.4, increased the percentage of OM from 1.2 to 3.5, as well as total nitrogen from 7 to 35 mg kg-1 and available phosphorus from 5 to 25 mg kg-1.

In contrast, the biosolid from the soap industry increased the pH from 8.1 to 8.8, did not modify the OM, only contributed 5 mg kg-1 of total nitrogen and contributed an increase in available phosphorus to 20 mg kg-1. In biosolids:soil mixtures, at 30 days the seedlings of Vicia faba significantly increased the yield of biomass from 1.1 to 13.7 g, which corresponds to 1 145% more, and the length from 12.7 to 56.3 cm, which corresponds to 343%.

Cited literature

Alaoui, A. E.; Bechtaoui, N.; Benidire, L.; Gharmali, A. E.; Achouak, W.; Daoui, K.; Imziln, B. and Oufdou, K. 2019. Growth and heavy metals uptake by Vicia faba in mining soil and tolerance of its symbiotic rhizobacteria. Environment protection engineering. 45(1):84-96. https://doi.org/ 10.5277/epe190107 

Antille, L. D.; Sakrabani, R.; Tyrrel, S. F.; Minh, S. L. and Godwin, R. J. 2013. Characterisation of organomineral fertilisers derived from nutrient-enriched biosolids granules. Appl. Environ. Soil. Sci. 1-11 pp. doi.org/10.1155/2013/694597.

Ashekuzzaman, S. M.; Forrestal, P.; Richards, K. and Fenton, O. 2019. Dairy industry derived wastewater treatment sludge: Generation, type and characterization of nutrients and metals for agricultural reuse. Journal of Cleaner Production. 230:1266-1275. doi.org/10.1016/ j.jclepro.2019.05.025.

Belachew, K. Y. and Stoddard, F. L. 2017. Screening of faba beans (Vicia faba L.) accessions to acidity and aluminium stresses. Peer. J. 5, e2963. doi.org/10.7717/peerj.2963.

Bücker-Neto, L.; Sobral-Paiva, A. L.; Dorneles-Machado, R.; Augusto-Arenhart, R. and Margis-Pinheiro, M. 2017. Interactions between plant hormones and heavy metals responses. Genet. Mol. Biol. 40(1suppl.):373-386. doi.org/10.1590/1678-4685-GMB-2016-0087.

Corrêa-Martins, M. N.; Ventura-Souza, V. and Silva-Souza, T. 2016. Genotoxic and mutagenic effects of sewage sludge on higher plants. Ecotoxicol. Environ. Saf. 124:489-496. doi.org/10.1016/j.ecoenv.2015.11.031.

Flemming, C. A.; Pileggi, V.; Chen, S. and Lee, S. S. 2017. Pathogen survey of pulp and paper mill biosolids compared with soils, composts, and sewage biosolids. J. Environ. Qual. 46(5):984-993. doi:10.2134/jeq2016.12.0467.

Flores-Félix, E.; Moreno-Casillas, H.; Figueroa-Viramontes, U. and Potisek-Talavera, M. C. 2014. Disponibilidad de nitrógeno y desarrollo de avena forrajera (Avena sativa L.) con aplicación de biosólidos. Terra Latinoam. 32(2):99-105.

Ghouili, E.; Sassi, K.; Jebara, M.; Hidri, Y.; Ouertani, R. N.; Muhovski, Y.; Jebara, S. H.; Ayed, M.; Abdelkarim, S.; Chaieb, O.; Jallouli, S.; Kalleli, F.; M’hamdi, M.; Souissi, F. and Ghassen, A. 2021. Physiological responses and expression of sugar associated genes in faba bean (Vicia faba L.) exposed to osmotic stress. Physiol. Mol. Biol. Plants. 27(1):135-150. https://doi.org/10.1007/s12298-021-00935-1.

Gomiero, T.; Pimentel, D. and Paoletti, M. G. 2011. Environmental impact of different agricultural management practices: conventional vs. organic agriculture. Crit. Rev. Plant. Sci. 30(1):95-124. doi.org/10.1080/07352689.2011.554355.

González-Flores, E.; Ramos-Barragán, J. E.; Tornero-Campante, M. A. y Murillo-Murillo, M. 2017. Evaluación de dosis de biosólidos urbanos en maíz bajo condiciones en el invernadero. Rev. Mex. Cienc. Agric. 8(1):119-132. doi.org/10.29312/remexca.v8i1.76.

Goss, M. J.; Tubeileh, A. and Goorahoo, D. 2013. A review of the use of organic amendments and the risk to human health. In: Adv. Agron. Sparks D. J. (Ed). First edition. Academic Press. San Diego, CA, USA. 275-379 pp. https://doi.org/10.1016/B978-0-12-407686-0.00005-1

Gupta, K.; Srivastava, A.; Srivastava, S. and Kumar, A. 2020. Phyto-genotoxicity of arsenic contaminated soil from Lakhimpur Kheri, India on Vicia faba L. Chemosphere. 241:1-10. https://doi.org/10.1016/j.chemosphere.2019.125063.

Harris-Valle, C.; Mora-Guzmán, E.; Palafox-Rodríguez, M.; Pérez-Pacheco, C. K.; Mejía-Franco, V. y Vázquez-Flores, Y. 2019. Crecimiento de haba en simbiosis con microorganismos nativos de regiones productoras del norte de Puebla. Rev. Fitotec. Mex. 42(3):243-250.

Hernández, R. J. J.; Rivera, M. M. C. y Arellano, E. R. 2017. Análisis de biosólidos para una hortaliza. Joven en la ciencia. 3(2):240-344.

ICAMEX. 2023. Instituto de Investigación y Capacitación Agropecuaria, Acuícola y Forestal. El cultivo del haba. Gobierno del Estado de México. https://icamex.edomex.gob.mx/haba.

INIFAP. 2017. Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias. Agenda Técnica Agrícola de Estado de México. 423 p. https://vun.inifap.gob.mx/BibliotecaWeb/-Content?//=AT.

Lang, L. N. and Smith, S. R. 2008. Time and temperature inactivation kinetics of enteric bacteria relevant to sewage sludge treatment processes for agricultural use. Water Res. 42(8-9):2229-2241. doi.org/10.1016/j.watres.2007.12.001.

Mantilla, M. G.; Sandoval, Y. L.; Ramírez, C. E. M.; Gasca, A. S.; Navarro, F. J.; Hernández, C. N.; García, R. J. L.; Esquivel, S. A. y Calderón, M. C. G. 2017. Energía limpia del agua sucia: aprovechamiento de aguas residuales. Instituto mexicano de tecnología del agua. México, DF. 88 p.

Piršelová, B.; Kuna, R.; Lukáč, P. and Havrlentová, M. 2021. Effect of cadmium on growth, photosynthetis pigments, iron, and cadmium accumulation of faba bean (Vicia faba cv. Aštar). Agriculture (Poľnohospodárstvo). 62(2):72-79. https://doi.org/10.1515/agri-2016-0008.

Postisek-Talavera, T. M.; Figueroa-Viramontes, U.; González-Cervantes, J. I. y Orona-Castillo, I. 2010. Aplicación de biosólidos al suelo y su efecto sobre contenido de materia orgánica y nutrimentos. Terra Latinoam. 28(4):327-333.

Price, G. W.; Langille, G. I. M and Svetlana, N. Y. 2021. Microbial co-occurrence network analysis of soils receiving short and long term applications of alkaline treated biosolids. Science of the total environment. 751:141687. https://doi.org/10.1016/j.scitotenv.

Rajhi, I.; Moussa, B.; Neji, I.; Baccouri, B.; Chikha, M. B.; Chammakhi, C.; Amri, M.; Brouquisse, R. and Mhadhbi, H. 2020. Photosynthetic and physiological responses of small seeded faba bean genotypes (Vicia faba L.) to salinity stress: identification of a contrasting pair towards salinity. Photosynthetica. 58(1):174-185. https://doi.org/10.32615/ps.2019.152 

SAGARPA. 2022. Secretaría de Agricultura y Desarrollo Rural. Anuario estadístico de la producción agrícola. Servicio de Información Agroalimentaria y Pesquera (SIAP). https://nube.siap.gob.mx/cierreagricola/.

Sandaña, P.; Orena, S.; Santos, R. J.; Kalazich, J. and Uribe, M. 2018. Critical value of soil Olsen-P for potato production systems in volcanic soils of Chile. J. Soil. Sci. Plant. Nutr. 18(4):965-976. doi.org/10.4067/S0718-95162018005002801.

SEMARNAT. 2000. Secretaría del Medio Ambiente y Recursos Naturales. Norma Oficial Mexicana NOM-021-. Que establece las especificaciones de fertilidad, salinidad y clasificación de suelos, estudio, muestreo y análisis. Diario oficial de la federación. http://www.ordenjuridico.gob.mx/documentos/federal/wo69255.pdf.

SEMARNAT. 2002. Secretaría del Medio Ambiente y Recursos Naturales. Norma Oficial Mexicana NOM-004. Protección ambiental lodos y biosólidos especificaciones y límites máximos permisibles de contaminantes para su aprovechamiento y disposición final. http://biblioteca.semarnat.gob.mx/janium-bin/detalle.pl?Id=20200923160311.

Tang, L.; Hamid, Y.; Zehra, A.; Sahito Z. A.; Hec, Z.; Hussain, B.; Gurajala, H. K. and Yang, X. 2019. Characterization of fava bean (Vicia faba L.) genotypes for phytoremediation of cadmium and lead co-contaminated soils coupled with agro-production. Ecotoxicol. Environ. Saf. 171:190-198. https://doi.org/10.1016/j.ecoenv. 2018.12.083.

Vela, C. M. A.; López, T. Z. G.; Sandoval, C. E.; Tornero, C. M. A. y Cobos, P. M. A. 2018. La fertilización órgano-mineral en el rendimiento de haba en suelo e hidroponia en agricultura protegida. Rev. Mex. Cienc. Agríc. 9(8):1603-1614.

Velasco, M. A. 2010. Modificación del plan municipal de desarrollo urbano del municipio de Nextlalpan. Periódico oficial del gobierno del estado libre y soberano de México. Estado de México. 172 p.