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Article

Evaluation of Cytotoxic and Genotoxic Risk Derived from Exposure to Pesticides in Corn Producers in Tlaxcala, Mexico

by
Antonio Rivera
1,
Lilia Cedillo Ramírez
1,
Conrado Parraguirre Lezama
2,
Alfredo Baez Simon
2,
Beatriz Laug Garcia
3 and
Omar Romero-Arenas
2,*
1
Centro de Investigaciones en Ciencias Microbiológicas, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Ciudad Universitaria, Puebla CP72570, Mexico
2
Centro de AgroecologÍa, Benemérita Universidad Autónoma de Puebla, Edificio VAL 1, Km 1,7 Carretera a San Baltazar Tetela-Zacachimalpa, Puebla 72960, Mexico
3
Laboratorio de Ecología Molecular Microbiana, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Ciudad Universitaria, Puebla CP72570, Mexico
*
Author to whom correspondence should be addressed.
Appl. Sci. 2022, 12(18), 9050; https://doi.org/10.3390/app12189050
Submission received: 2 August 2022 / Revised: 1 September 2022 / Accepted: 2 September 2022 / Published: 8 September 2022
(This article belongs to the Special Issue Good Agricultural Practices: Application, Monitoring and Ecotoxicological Effects in Pesticide Use)
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Abstract

:
Corn cultivation represents the largest type of agricultural production in Mexico, with great economic, social, and cultural importance. The health of corn producers could be compromised by the extensive and accumulated use of pesticides. The effects of pesticides in terms of their cytotoxic and genotoxic damage in two groups of peasant maize producers in Tlaxcala, Mexico, were considered here. The buccal micronucleus cytome assay was used as an indicator of cytotoxicity and genotoxicity, along with nuclear abnormalities present in farmers who had used pesticides in the last thirty years. In total, 21 commercial products used in corn production were identified, mainly herbicides belonging to the chlorophenoxy, triazine, and organophosphate compounds; in addition, it was observed that a small group of farmers use the active ingredient carbofuran, as well as insecticides and fungicides. The results show that farmers with higher pesticide use present higher rates of cytotoxic and genotoxic damage compared to the group of producers with higher incidence rates of agroecological practices and lower rates of pesticide use, as revealed by the micronucleus assay, as well as by nuclear abnormalities present in the epithelial cells of the buccal mucosa. The agroecological farmer group used only herbicides, with the 2,4-D (Hierbamina) being the greatest use in maize cultivation.

1. Introduction

Maize (Zea mays L.) is the second most important cereal crop worldwide [1]. Corn consumption represents an excellent source of protein, carbohydrates, fiber, amino acids, and folic acid, as well as minerals such as potassium, magnesium, sodium, and calcium [2], being the most important seed in the Mexican diet [3]. Mexico ranks seventh as a maize producer globally, producing 17,910,944 tons annually; however, it operates as an importer of the same cereal to meet domestic demand [4]. The area planted with maize in the state of Tlaxcala, Mexico, for the year 2021 measured 11,710 ha, producing 36,761 tons and ranking fourteenth in national production [5].
The use of harmful chemical substances in the environment has become a topic of interest and great debate in recent decades [6]. Pesticides are widely applied in agriculture for protection purposes against a wide variety of unwanted living organisms, which can cause large economic losses. It should be mentioned that in 2019 approximately 2 million tons of pesticides were used, of which 47.5% were herbicides, 29.5% were insecticides, 17.5% were fungicides, and 5.5% were other pesticides from around the world [7]. It has been determined that pesticides have negative effects on the population that are directly related to agricultural activities, particularly in developing countries, representing a threat to human health, with more than 900 different pesticides in the market, covering 2600 products. [6].
The World Health Organization (WHO) classifies pesticides according to their toxicity, grouping them as extremely hazardous pesticides, highly hazardous pesticides, moderately hazardous pesticides, slightly hazardous pesticides, and pesticides that are unlikely to present an acute hazard [6,8]. Paying attention to pesticide formulations, most of the commonly used formulations contain pyrethroids, organophosphates, organochlorines, carbamates, glyphosate, glufosinate, and triazoles [9].
In Mexico, the technological schemes of the green revolution were promoted in the second half of the 20th century, incorporating conventional practices, fertilizers, and pesticides in corn production systems [10,11]. The consumption of pesticides per hectare showed an increase from 1940 to the last decade of the 20th century. In this context, it is estimated that there are around 900 pesticides used by producers in the country, with chemical insecticides being the most recurrent for the production of corn, cotton, potatoes, peppers, tomatoes, beans, wheat, avocado, coffee, and tobacco, in quantities ranging from 395 to 13,163 tons per year [12].
The genotoxic potential is a primary risk factor for long-term effects such as carcinogenic and reproductive toxicology, as well as degenerative diseases [13]. Genotoxicity tests assess at the DNA level single and double strand breaks, point mutations, chromosomal aberrations, and micronucleus formation, among others [14]. Biomonitoring studies focused on genomic modifications have been carried out in pesticide-exposed populations from different countries to elucidate the risks associated with exposure to specific compounds or classes of compounds or specific cultivation practices [15]. Among them, the buccal micronucleus cytome assay (BMCA) has been used in the last decades, using peripheral lymphocytes or exfoliated cells from the buccal mucosa [13]. Micronuclei are chromosomal fragments or complete chromosomes that are formed during the anaphase; these fragments cannot be included in the nuclei of daughter cells but form single or multiple micronuclei in the cytoplasm [16].
The BMCA of buccal cells is useful as a biomarker of the genetic damage caused by lifestyle habits such as the consumption of tobacco products and alcohol, micronutrient deficiencies, exposure to pesticides, as well as inherited genetic defects [17]. The studies by Marcelino et al. [18] indicated morphological changes of 1.11 alterations per thousand cells in farmers not exposed to pesticides compared to a damage rate of 3.28 alterations per thousand cells in growers using pesticides. Balderrama-Carmona et al. [19] demonstrated that 53% of agricultural workers show cell damage upon exposure to herbicides such as glyphosate, aminomethylphosphonic acid (AMPA), and picloram via cell proliferation and micronucleus evaluations.
The non-invasive nature of this technique makes it an attractive candidate for the biomonitoring of human populations or individuals [20]. Therefore, the main objective of this study was to compare whether there are differences in relation to the cytotoxic and genotoxic effects in two groups of corn producers belonging to the municipality of Vicente Guerrero, who make use of different pesticides, using a detailed description of the types of nuclear abnormalities present in both groups.

2. Materials and Methods

2.1. Study Area

The locality of Vicente Guerrero is located at kilometer six of the Nanacamilpa highway, within the municipality of Españita, in the western part of the state of Tlaxcala (Figure 1). Españita is in the central highlands of Mexico between parallels 19°22′ N and 19°30′ N in latitude and meridians 98°20′ W and 98°31′ W in longitude, at an altitude range of 2400 to 2900 masl. It is bordered to the north by the municipality of Nanacamilpa de Mariano Arista, to the east by the municipality of Hueyotlipan, to the south by Ixtacuixtla de Mariano Matamoros, and to the west by the state of Puebla. Españita represents 3.5% of the state’s surface area and is made up of 45 localities [21]. The study community has a temperate subhumid climate (Cw) and an average rainfall rate of 800 mm [22]. The rainfed maize crop represents 64.08% of total the agricultural area planted in the municipality [23].

2.2. Sample Size Selection

The participants considered for this research are composed solely of male corn producers residing in the municipality of Vicente Guerrero, Españita-Tlaxcala. Using a simple random sampling design considering a bilateral alpha (d) of 0.05, a normal distribution (Z) of 1.96, and a coefficient of variability (CV) of 0.8, we calculated the sample (n) with respect to the list of local producers (N = 90) according to the formula indicated by Santoyo [24] in Equation (1):
n = (NZ2 × CV2)/(((N − 1)d2) + (Z2 × CV2))
where N represents the total number of producers, Z is the normal distribution, CV is the coefficient of variability, d is the bilateral alpha, and n is the sample size.
A questionnaire on exposure to pesticides and sociodemographic characteristics was applied to a total of 90 agricultural workers, called “Campesinos”, with thirty years of experience to obtain general information, in addition to assessing the current use and exposure to the most used pesticides in the production of maize and determining the toxicity according to the relationship with the list of highly dangerous pesticides [25]. The voluntary participants were carefully selected to represent the group with the highest exposure to the use of pesticides (i.e., the group of conventional farmers or FC), identifying 47 farmers producing corn. The control group contained 43 farmers with a higher frequency of agroecological practices or FA and with less use of pesticides. This sample size was sufficient to detect sample effect sizes of approximately two-thirds of the standard deviation.
The selection of the study areas and the recruitment of the participants for both groups “peasant producers of corn” were carried out with the support of the peasant organization Grupo Vicente Guerrero (GVG), which in recent years has been acting as a spokesperson for the peasants, with the purpose of consolidating agroecological alternatives to face poverty and environmental deterioration. They actively work in the municipalities of Nanacamilpa, Españita, Ixtacuixtla, Tlahuapan and Tepetitla, and Ixtenco, carrying out activities related to sustainable agriculture, pesticide use reduction, food sovereignty, and gender equity [26].

2.3. Buccal Micronucleus Cytome Assay (BMCA)

Authorization was obtained from the ethics committee of the Benemerita Universidad Autonoma de Puebla, Mexico, and the Programa de Posgrado en Manejo Sustentable de Agroecosistemas (FACMED/CI2018-2020) in accordance with the Declaration of Helsinki. Prior to the non-invasive examination, the maize farmers were informed about the methods and procedure to be performed. The participants in both groups rinsed their mouths twice with tap water to remove possible food residues, then gently scraped the oral mucosa from both cheeks with separate sterile wooden spatulas, according to the procedure described by Tolbert et al. [27].
For each participant, the right and left cheek slides were prepared and transported to the laboratory for analysis. The smears were immersed in 5 mL of saline (0.9% NaCl) for 5 min and then subjected to centrifugation for 10 min at 1500 rpm [28]. The supernatant was discarded while the cell pellet was spread on a clean glass slide and fixed in freshly prepared cold methanol (3:1 glacial acetic acid mixture) for 15 min, then subsequently stained using the Feulgen reaction technique [29]. The stained slides were observed under oil immersion using an optical microscope (Carl Zeiss, Jena, Germany) at 10× and 40× magnification to identify and record micronucleate cell (MN) counts and nuclear abnormalities.
The frequency of micronucleated cells (MN), total number of micronuclei (MNi), lobulated nuclei “broken eggs” (BUD), and binucleated cells (BN) were determined as indicators of possible aneuploidy and genomic instability. In addition, indicators of genotoxic effects, such as cells with condensed chromatin (CC), karyorrhectic cells (KR), karyolytic cells (KL), pyknosis (PY), and basal cells (BC), were analyzed to assess the proliferative activity in the epithelium of the buccal mucosa [28]. One thousand epithelial cells were examined for everyone in both groups. All parameters were classified according to Bolognesi and Fenech [30] and Holland et al. [31].

2.4. Statistical Analysis

With the information obtained from the surveys, a database was created to determine the centrality, dispersion, and association of the factors (means, frequencies, and proportions). The normality of the variables was confirmed by the Kolmogorov–Smirnov test. Chi-square (χ2) tests were used to evaluate the association between demographic factors, knowledge, attitude towards smoking and alcoholism, and preventive measures with regard to the total number of micronuclei (MNi) present in the study subjects. Subsequently, the Mann–Whitney U test (p ≤ 0.05) was performed to determine statistically significant differences in nuclear abnormalities (total number of micronuclei (MNi), condensed chromatin (CC), lobulated nucleus (BUD), micronuclei (MN), karyolytic cells (KL), karyorrhectic cells (KR), pyknosis (PY), basal cells (CB). and binucleated cells (BN) between the exposed group (FC) and the control group (FA) per 1000 cells.
Finally, a multivariate analysis was performed and the generalized linear model was used, which was adjusted to control the factors of age, educational level, the confounding factors “smoking and alcoholism”, and protection (use of long sleeves, use of glasses, use of protectors to nose, use of mask, use of gloves, and use of boots) with respect to the total number of micronuclei (MNi) present in peasant corn producers from the town of Vicente Guerrero in the municipality of Españita, Tlaxcala, Mexico. The adjusted odds ratio (AOR) was calculated to assess the strength of the association between the outcome and the explanatory variables. The significance of the statistical associations was confirmed by odds ratios with a 95% confidence interval (CI) and p ≤ 0.05 values. All calculations were performed with the statistical package SPSS Statistics version 17 for Windows.

3. Results

The sociodemographic characteristics of both study groups were comparable. The ages of the producers ranged between 47 and 59 years with statistically similar ages (FC: X = 42.86 ± SD = 14.65; FA: X = 40.62 ± SD = 15.83). The level of schooling was predominantly elementary education for the FC group (57.44%), while the FA group had a higher educational level (secondary education, 51.16%). It was also observed that the alcoholic habit prevailed in both groups, unlike the tobacco habit, where 78.72% of the peasants were non-smokers for the FC group, being higher in the FA group (Table 1).
Currently, most peasants plant between one and five hectares (78.8%); few peasants cultivate more than 15 hectares (4.5%). The predominant type of system is maize monoculture (59.1%), although the “milpa system” was detected less frequently (2.5%), which refers to a traditional agricultural system made up of a polyculture, where the main species is corn, accompanied by various species of beans, pumpkins, and chili peppers, which constitutes a dynamic space of genetic resources. It should be noted that of the 90 maize-producing peasants, 34.04% (16/47) of the FC group and 58.13% (25/43) of the FA group reported having received training in pesticide management.
The FC group reported higher pesticide exposure with a 6 day, 8 h work week, with thirty years of experience in corn production. The FA group carried out various agroecological practices in the production of corn, with a higher frequency of mixed activities and less exposure to pesticides, with a work week of 7 days at 10 h a day, presenting the same experience of years worked as the FC group. According to the information obtained from the corn farmers surveyed here, 99.7% of the FC group had used some type of chemical pesticide, applying it two to three times per agricultural cycle. In the case of the FA group, it was identified that a small percentage of farmers used chemical pesticides (2.32% to 18.60%), applying them at least once during the agricultural cycle. In total, 21 commercial products were identified, mainly herbicides and to a lesser extent insecticides and fungicides (Table 2).
The group of peasant maize producers in the FC group reported using more herbicides, such as Esteron, Hierbamina, and Gesaprim, followed by the group of insecticides (Arrivo and Karate) and to a lesser extent the fungicide, with the commercial brand Folicur being the one that showed the greatest use in the cultivation of maize by the peasants.
The results showed that in the FA group, only the use of herbicides was identified, with the commercial brand Hierbamina being the one that presented more use in the maize crop (18.60%). It should be noted that its application is conditioned by the presence of insects, weeds, or microorganisms that damage the maize crop, which means that one, two, or even three applications are required per agricultural cycle. The reading of labels is a practice that is not carried out regularly, since a little more than one-third of the peasant maize producers surveyed in the FA group (37.9%) stated that they always read their contents. In the case of the group exposed to pesticides (FC), it was observed that 52.2% of the peasant did not understand the interpretation of the yellow color band (moderately dangerous products), while 32.8% did not understand the interpretation of the blue color band (slightly dangerous products); the opposite was the case with the red color label (extremely dangerous), showing that 80% of the peasant maize producers identified what this color represents.
The total numbers of micronuclei (MNi) presented highly significant differences (p ≤ 0.05) between the corn producers of the FC group (0.408 ± 0.135) with respect to the control group (FA), who had a higher incidence rate of using agroecological practices (0.253 ± 0.221). The highest frequency of micronuclei (MNi) occurred in the range of 47–59 years for both groups; however, the highest amount of MNi was observed in the older individuals for both groups (Table 3).
An intergroup (between groups) comparison of the total numbers of micronuclei revealed statistically significant differences (p ≤ 0.05) between corn farmers in the FC group (exposed to chemical pesticides) and the FA control group.
There were highly significant differences for the educational factor with secondary preparation for the FA group with respect to the other educational sections for both groups, presenting a lower amount of MNi. However, the corn-producing peasants without education were the ones who presented the highest amounts of MNi in both groups. The smoking factor presented statistically significant differences, whereby the FC group presented a greater amount of MNi in the ranges of more than 10 cigarettes/day and 1–5 cigarettes/day compared to the FA group. However, for the alcohol consumption factor, the FA group presented a greater amount of MNi for the range of 4 or more times a week.
Finally, the personal protection factor that integrated 3 to 5 measures used by the producers presented highly significant differences (Table 3).
The buccal mucosal cytome assay used to evaluate cells with nuclear abnormalities and determine the potential risk and genotoxic damage from the exposure to pesticides in maize farmers showed highly significant differences in both groups (Table 4).
The highest mean frequency values of nuclear abnormalities in buccal epithelial cells were presented in the FC group in relation to the FA group, and the highest frequencies of abnormalities corresponded to total micronuclei (MNi), condensed chromatin (CC), karyorrhexis (KR), and micronuclei (MN), followed by pyknotic nuclei (PY) and lobulated nuclei (BUD).
In the present study, it was observed that for degenerative nuclear changes related to genotoxic damage, such as karyolysis (KL), binucleated cells (BN), and basal cells (CB), there were no significant differences for both groups, as shown in the Figure 2.
When the sociodemographic characteristics of age, educational level, habits (alcohol consumption and smoking), and personal protection were evaluated with the total micronuclei (MNi) in both groups using the ordinal logistic regression model (Table 5), they showed a significant association, indicating that the explanatory power is very strong.
The corn producers of the FC group were twice as likely to have a greater number of total micronuclei (AOR = 1.661, 95% Cl: 1.306–2.112, p ≤ 0.001); however, the other variables did not present statistical differences according to their categories for each level in comparison (p = 0.759, 0.659, 0.610, 0.893 for age, educational level, and habits of alcohol consumption and smoking, respectively).
The maize producers from the FA group had an 11.60% chance of presenting fewer micronuclei (AOR = 0.884, 95% Cl: 0.874–0.903, p ≤ 0.003), unlike the FC group. On the other hand, the probability of presenting micronuclei increased if the corn producers did not use some type of personal protection when applying the pesticides. However, the use of protection measures may be conditioned by inconvenience, ignorance, and the notification of a health risk. The producers expressed difficulty breathing with the use of masks and not having the proper equipment. They also preferred to use alternative materials such as plastic bags to cover their backs and a handkerchief to cover their mouths and noses when they applied pesticides on their plots. In addition, most of the producers interviewed from both groups do not use protective equipment when using pesticides.
The results clearly demonstrated that higher pesticide exposure in the FC group caused changes in cytokinesis and led to a higher number of cells with degenerative nuclear changes related to genotoxic damage, as can be seen in Figure 3.

4. Discussion

Within the scope of our study, we can point out that the average age of the corn-producing peasants in the Vicente Guerrero locality, belonging to the municipality of Españita, agreed with the study carried out by Damián-Huato and Ramírez-Valverde [32] for the state of Tlaxcala, Mexico. Furthermore, according to the latest National Agricultural Survey (ENA), 45.8% of producers immersed in agricultural activities are considered older adults [23].
It was determined that the peasant maize producers had a low educational level, given that the majority concluded their education with primary studies (57.45%) for the FC group, followed by the FA group with a higher incidence of agroecological practices and less use of pesticides and with secondary school completion (51.16%). Similar situations also occurred in the studies by Carbajal-López et al. [33] and Ramírez-Mora et al. [34]. Regarding the use of pesticides among peasant maize producers, Damian-Huato and Ramirez-Valverde [35] estimated the use of herbicides at 57%, followed by 67.5% for insecticides, with the commercial brand Karate being the most recurrent among peasants belonging to the state of Tlaxcala, Mexico.
Sánchez and Romero [26] reported that 6% of conventional corn monoculture farmers used hybrid seeds, 93% applied fertilizers, and 68% used herbicides, unlike traditional corn polyculture farmers, where 3% of the peasants used hybrid seeds, 91% used fertilizers, and 11% applied herbicides, which were similar results to those found in the present investigation, with the Hierbamina commercial brand being the one with the greatest representation in the FA group. It is worth mentioning that for the present study there is a considerable use of herbicides and insecticides (84.8% and 81.8%, respectively), which generates a greater dependence on agrochemical inputs for the group with the highest use of pesticides (FC). Nicholson and Williams [36] stated that each year 2700 million kg of pesticides are applied in agriculture worldwide. These chemical synthesis products, including insecticides, herbicides, and fungicides, when applied in corn production, directly threaten the health of farmers [37]. This fact can be interpreted as an indication that the use of pesticides is directly related to a toxicological profile that causes DNA damage and long-term cytotoxic effects [38].
The buccal micronucleus cytome assay (BMCA) is considered a suitable test for the evaluation of risks associated with chemical induction [39,40], which can be used to detect cytotoxic and genotoxic damage caused by short- and long-term exposures to pesticides with effects on the oral mucosa in humans [41]. The peasant maize producers directly involved in the handling and periodic application of pesticides are at high risk of cytotoxic and genotoxic damage, especially due to the absence of protective equipment or not using it properly [42]. In addition, a low level of knowledge about the color code of hazardous pesticides was presented by the peasants in both groups, which is common in many agricultural countries where pesticides are used in food production [43].
The results of the present investigation show that the total micronuclei (MNi) can be considered as markers of genotoxicity present in the group with the highest use of pesticides (FC), since they presented significantly higher frequencies compared to the control group (FA). This corroborates the findings of a similar and recent study on banana cultivation in Brazil [44], which indicated a significant increase in micronuclei in the pesticide-exposed group over the control group.
The frequency analysis of nuclear anomalies showed significantly higher rates in the peasant maize producers exposed to pesticide use (FC) compared to the control group (FA). The highest frequencies were observed in cells with CC, MN, PY, BUD, and KR, a similar situation to that reported by Benedetti et al. [45]. According to Bolognesi et al. [39], these factors are mainly related to cell death and chromosomal instability processes. As in our study, Dutta et al. [46] observed a higher proportion of nuclear abnormalities and cell death parameters in tea garden workers exposed to pesticides.
Regarding the frequency of cells with condensed chromatin (CC), the values obtained were high for both groups (560 ± 251/1000 cells observed vs. 390 ± 165/1000 cells observed) compared to the values recorded in other investigations [33,45,47]; however, studies such as those by Martinez-Valenzuela et al. [48] and Ortega-Martínez et al. [49] reported frequencies similar to those obtained for the exposed group (FC), in the first case involving pesticide applicator pilots, and in the second case involving greenhouse day laborers; both groups had been exposed for more than 25 years, working with complex mixtures and with little use of protective equipment.
Regarding the use of pesticides and according to what was reported by the peasants from both groups, the use of compounds applied for the production of maize is based mainly on the chemical family of chlorophenoxy, triazines, and pyrethroids, which coincides with the producers of maize and soybean from the Province of Córdoba, Argentina [50]; however, the use of organochlorine, organophosphate, and carbamate pesticides continues to be reported more frequently in Mexico, which is why it ranked third in the purchase of agrochemicals in the largest market in North America, growing by 5.2% during the period 2017–2022 [48,51,52].
Based on the results obtained here, it was detected that the group of exposed peasants (FC) presented a greater rate of pesticide use, due to the agricultural conditions developed in the study locality for the production of corn, as it is a predominantly rainfed crop [53]. In addition, the combination of at least two products with different active ingredients during the agricultural cycle exposes farmers to more toxic mixtures that increase the risk to health and can favor the presence of cytogenetic alterations, as shown by the study by Aiassa et al. [54], where they positively correlated this factor with the presence of cytogenetic alterations in agricultural workers with a minimum exposure of three years. It should be noted that some of these active ingredients are presented in Table 2 and are classified in the World Health Organization database [55] as moderately hazardous and highly hazardous.
In many investigations, age is a factor that has been significantly correlated with the presence of damaged cells in workers exposed to pesticides [50]. The results obtained in the present study showed a significant difference in the numbers of total micronuclei (MNi) for both groups; likewise, it was observed that the producers with advanced ages (≥60 years) presented the highest number of total micronuclei (MNi). Balderrama-Carmona et al. [19] reported that age does not influence the increase in micronuclei (MN), since they did not find cell damage in samples of agricultural workers with 30 and 45 years of exposure to pesticides, results that contrast with those presented in the present investigation and agree with those obtained by Ferraz et al. [56], who pointed to the variable “age” as the main factor associated with the induction of damage to the genetic material. It is worth mentioning that the genotoxic and cytotoxic effects related to the “age” variable can be attributed to specific components of the pesticides, which can sometimes have different effects [57], since agricultural workers are generally exposed to a mixture of different pesticides. Likewise, Carbajal et al. [33] mentioned that the increase in MNi could be attributed to a further progressive deterioration in the repair capacity and an increase in free radicals in cells.
Alcoholic and smoking habits were present in both groups, characteristics that are similar to those reported by Matheus et al. [58], where they showed a lower prevalence of micronuclei (MN) for the smoking habit (21.9% and 19.5%) in relation to the alcoholic habit (68.3% and 58.5%). When analyzing the statistical association of the total micronuclei (MNi) with the aforementioned habits (confounding factors), no statistically significant relationship was found, which show that these variables do not directly influence either group of farmers involved in the present study. However, the use of pesticides used by peasant corn producers (Table 2) can have negative effects on their health, coinciding with the results obtained by Larrea et al. [33], Carbajal et al. [59], and Tomiazzi et al. [60]
For the personal protection variable, statistically significant differences were found in the peasants exposed to the use of pesticides (FC) compared to the group with the highest incidence of agroecological practices and the least use of pesticides (FA). Among the most appropriate protective equipment items are gloves, boots, and raincoats [47,48,50]. However, when analyzing the statistical association of the total micronuclei (MNi) with the personal protection variable, we found highly significant differences, which may have been due to the fact that most of the corn producers in the FC group (40.10%) do not use any measure of protection, among which the use of masks and goggles stands out, so they are more exposed and have a greater risk towards their health due to genotoxic damage [54].
The results obtained in the present investigation show that there was a significant statistical difference in both groups for the personal protection variable, which integrated 3 to 5 security measures used by the producers. We can explain this relationship by citing an example found for both groups. For example, the backpack-type manual sprinkler is the most cited piece of equipment among farmers in charge of extensions of less than 5 hectares [34]. Such equipment use predisposes the users to a higher risk of damage due to inhalation and the percutaneous absorption of pesticides, mainly if the appropriate personal protective equipment is not used [61]. According to Lesmes-Fabian et al. [62], the most exposed parts of the body when using this equipment are the arms, back, thighs, and legs, with direct contact due to spills when the application equipment is not in good condition. In the present investigation, it was possible to appreciate that there is a greater application rate of pesticides when there is no wind in the group of peasants with a greater frequency of agroecological practices (FA). Ferré et al. [47] considered the application of pesticides when there is no wind as a good agricultural practice, as observed in the present investigation. However, the lack of training in its handling and the use of a protective mask in both groups, as well as the absence of protective glasses, expose corn producers to a high risk to their health [63]. The conclusions of the various epidemiological studies on the occupational exposure of agricultural workers to pesticides are not unequivocal. The International Agency for Research on Cancer (IARC) has classified certain pesticides as genotoxic in humans, such as the herbicides glyphosate, malathion, and their formulations [20], which were reported by the producers of the FC group in our study.
Joco et al. [64] mentioned that there is a risk factor if there is a significant correlation with the presence of damage to the genetic material caused by exposure to pesticides, as can be seen in Figure 2. According to Guzmán-Plazola et al. [65], the empirical knowledge acquired by the agricultural workers is transmitted from father to son, including the mishandling of these products, which in most cases affects the health of the farmers. Consequently, the risk to the health of the producers exposed to the greater use of pesticides in the Community of Vicente Guerrero, Tlaxcala, Mexico, may increase if they do not consider using adequate protective equipment for the application of pesticides during corn cultivation.

5. Conclusions

Within the framework of this study, samples of epithelial cells collected under field conditions showed that cytotoxic biomonitoring and genetic damage in the community of Vicente Guerrero, Tlaxcala, Mexico, are useful and feasible tools to estimate the health risks to corn farmers derived from exposure to chemical pesticides.
The peasant corn producers with higher usage rates of pesticides (FC) had a higher risk of presenting genotoxic and cytotoxic damage compared to the group of farmers with a higher incidence of agroecological practices and less use of pesticides (FA), as revealed by the testing of micronuclei, as well as nuclear abnormalities present in cells of the epithelium of the buccal mucosa. This exposes the need to evaluate the secondary effects on the health of farmers to prevent diseases related to the continuous exposure to chemical synthesis products.
The types of pesticides most frequently reported by the peasants exposed to pesticides (FC) in maize cultivation are herbicides belonging to the chlorophenoxy and triazine chemical groups, classified mainly as moderately hazardous, while a small group of peasants use the active ingredient carbofuran, classified as highly dangerous. The FA group presented the use of herbicides, with Hierbamina (2,4-D) being the one with the greatest use in maize cultivation.
The results presented in this research show the need for protection measures for farmers exposed to higher use of pesticides (FC). Therefore, we assume a serious lack of knowledge about the need for protection measures, as well as a general neglect of security measures and regulatory provisions. It is urgently recommended to implement training, education, and information programs on the proper use of pesticides, as well as the provision of protective clothing and supervision by local authorities.

Author Contributions

Conceptualization, O.R.-A., A.R. and L.C.R.; methodology, A.R., L.C.R. and O.R.-A.; software, C.P.L. and O.R.-A.; validation, L.C.R., A.R. and O.R.-A.; formal analysis, A.B.S., C.P.L. and O.R.-A.; resources, A.R. and L.C.R.; original—draft preparation, A.B.S., B.L.G. and O.R.-A.; writing—review and editing, A.R. and O.R.-A.; visualization, O.R.-A. and A.B.S.; supervision, L.C.R.; project administration, O.R.-A.; funding acquisition, A.R. All authors have read and agreed to the published version of the manuscript.

Funding

This research was supported by the program PRODEP 2020 of the Secretaría de educación Pública of Mexico (SEP); the Consejo Nacional de Ciencia y Tecnología (CONACyT), number 927199; and Benémerita Universidad Autónoma of Puebla, number 100233566.

Institutional Review Board Statement

The study was conducted in accordance with the Declaration of Helsinki and was approved by the ethics committee of the Benemerita Universidad Autonoma de Puebla, Mexico, and the Postgraduate Program in Sustainable Management of Agroecosystems (FACMED/CI2018-2020) for studies with human beings.

Informed Consent Statement

Informed consent was obtained from all subjects involved in the study.

Acknowledgments

The authors are grateful to Consejo Nacional de Ciencia y Tecnología (CONA-CyT) and Laboratory 204 of the Center for Agroecology at Benémerita Universidad Autónoma de Puebla, México, as well as to the Vicente Guerrero Group and the authorities and producers of the town of Vicente Guerrero, Españita, Tlaxcala, for the facilities used to carry out this investigation.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Walne, C.H.; Reddy, K.R. Temperature Effects on the Shoot and Root Growth, Development, and Biomass Accumulation of Maize (Zea mays L). Agriculture 2022, 12, 443. [Google Scholar] [CrossRef]
  2. Siyuan, S.; Tong, L.; Liu, R. Fitoquímicos del maíz y sus beneficios para la salud. Food Sci. Hum. Wellness 2018, 7, 185–195. [Google Scholar] [CrossRef]
  3. Ruiz-Huerta, E.A.; Armienta-Hernández, M.A.; Dubrovsky, J.G. Bioaccumulation of heavy metals and as in maize (Zea mays L) grown close to mine tailings strongly impacts plant development. Ecotoxicology 2022, 31, 447–467. [Google Scholar] [CrossRef] [PubMed]
  4. Organización de las Naciones Unidas para la Alimentación y la Agricultura (FAO). Available online: https://www.fao.org/faostat/es/#data/QCL (accessed on 21 May 2022).
  5. Servicio de Información Agroalimentaria y Pesquera (SIAP). Available online: https://nube.siap.gob.mx/gobmx_publicaciones_siap/pag/2020/Atlas-Agroalimentario-2020 (accessed on 21 May 2022).
  6. Damalas, C.A.; Eleftherohorinos, I.G. Pesticide exposure, safety Issues, and risk assessment indicators. Int. J. Environ. Res. Public Health 2011, 8, 1402–1419. [Google Scholar] [CrossRef]
  7. Cech, R.M.; Jovanovic, S.; Kegley, S.; Koen Hertoge, K.; Leisch, F.; Zaller, J.G. Reducing overall herbicide use may reduce risks to humans but increase toxic loads to honeybees, earthworms, and birds. Environ. Sci. Eur. 2022, 34, 1–20. [Google Scholar] [CrossRef]
  8. Li, Z.; Jennings, A. Worldwide regulations of standard values of pesticides for human health risk control: A Review. Int. J. Environ. Res. Public Health. 2017, 14, 826. [Google Scholar] [CrossRef]
  9. Lucia, A.; Guzmán, E. Emulsions containing essential oils, their components or volatile semi chemicals as promising tools for insect pest and pathogen management. Adv. Colloid Interface Sci. 2021, 287, e102330. [Google Scholar] [CrossRef]
  10. Harwood, J. Could the adverse consequences of the green revolution have been foreseen? How experts responded to unwelcome evidence. Agroecol. Sustain. Food Syst. 2020, 44, 509–535. [Google Scholar] [CrossRef]
  11. Del-Val, E.; Ramírez, E.; Astier, M. Comparison of arthropod communities between high and low input maize farms in Mexico. CABI. Agric. Biosci. 2021, 2, 40. [Google Scholar] [CrossRef]
  12. García-Gutiérrez, C.; Rodríguez-Meza, G.D. Problemática y riesgo ambiental por el uso de plaguicidas en Sinaloa. Ra Ximhai 2012, 8, 1–10. [Google Scholar] [CrossRef]
  13. Bolognesi, C.; Creus, A.; Ostrosky-Wegman, P.; Marcos, R. Micronuclei and pesticide exposure. Mutagenesis 2011, 26, 19–26. [Google Scholar] [CrossRef] [PubMed]
  14. Reddy, M.V.; Storer, R.D.; Laws, G.M.; Armstrong, M.J.; Barnum, J.E.; Gara, J.P. Genotoxicity of naturally occurring indole compounds: Correlation between covalent DNA binding and other genotoxicity tests. Environ. Mol. Mutagen. 2002, 40, 1–17. [Google Scholar] [CrossRef] [PubMed]
  15. Sánchez-Alarcón, J.; Milić, M.; Kašuba, V.; Tenorio-Arvide, M.G.; Montiel-González, J.M.R.; Bonassi, S.; Valencia-Quintana, R. A Systematic Review of Studies on Genotoxicity and Related Biomarkers in Populations Exposed to Pesticides in Mexico. Toxics 2021, 9, 272. [Google Scholar] [CrossRef] [PubMed]
  16. Fenech, M.; Kirsch-Volders, A.T.; Natarajan, J.; Surralles, J.W.; Crott, J.; Parry, H.; Norppa, D.A.; Eastmond, J.D.; Tucker, P. Molecular mechanisms of micronucleus, nucleoplasmic bridge and nuclear bud formation in mammalian and human cells. Mutagenesis 2011, 26, 125–132. [Google Scholar] [CrossRef]
  17. Costa, S.; Costa, C.; Madureira, J.; Valdiglesias, V.; Teixeira-Gomes, A.; Guedes de Pinho, P.; Laffon, B.; Teixeira, J.P. Occupational exposure to formaldehyde and early biomarkers of cancer risk, immunotoxicity and susceptibility. Environ. Res. 2019, 179 Pt A, 108740. [Google Scholar] [CrossRef]
  18. Marcelino, A.F.; Wachtel, C.C.; Ghisi, N.d.C. Are Our Farm Workers in Danger? Genetic Damage in Farmers Exposed to Pesticides. Int. J. Environ. Res. Public Health 2019, 16, 358. [Google Scholar] [CrossRef]
  19. Balderrama-Carmona, A.P.; Valenzuela, M.; Zamora-Álvarez, L.A. Herbicide biomonitoring in agricultural workers in Valle del Mayo, Sonora Mexico. Environ. Sci. Pollut. Res. 2020, 27, 28480–28489. [Google Scholar] [CrossRef]
  20. Podrimaj-Bytyqi, A.; Borovečki, A.; Selimi, Q. The frequencies of micronuclei, nucleoplasmic bridges and nuclear buds as biomarkers of genomic instability in patients with urothelial cell carcinoma. Sci. Rep. 2018, 8, 17873. [Google Scholar] [CrossRef]
  21. Instituto Nacional de Estadística, Geografía e Informática (INEGI). Compendio de Información Geográfica Municipal 2010 Españita, Tlaxcala. Available online: http://geoweb2.inegi.org.mx/compendiosmun/ctrlpant (accessed on 12 October 2021).
  22. García, E. Modificaciones al Sistema de Clasificación Climática de Köppen, 5th ed.; Instituto de Geografía, Universidad Autónoma de México: México City, Mexico, 2004; pp. 11–90. [Google Scholar]
  23. Instituto Nacional de Estadística, Geografía e Informática (INEGI). Encuesta Nacional Agropecuaria 2019. Available online: https://www.inegi.org.mx/programas/ena/2017/ (accessed on 12 October 2021).
  24. Santoyo, H.; Ramírez, P.; Suvedi, M. Manual para la Evaluación de Programas de Desarrollo Rural. 2002. Available online: http://ciestaam.edu.mx/libro/manual-la-evaluacion-programas-desarrollo-rural-2/ (accessed on 11 April 2022).
  25. Pesticide Action Network (PAN). PAN International Consolidated List of Banned Pesticides 4th. Available online: http://pan-international.org/pan-international-consolidated-list-of-banned-pesticides/ (accessed on 12 May 2022).
  26. Sánchez, M.; Romero-Arenas, O. Evaluación de la sustentabilidad del sistema milpa en el estado de Tlaxcala, México. Rev. Col. San Luis 2018, 8, 107–134. [Google Scholar] [CrossRef]
  27. Tolbert, P.E.; Shy, C.M.; Allen, J.W. Micronuclei and other nuclear anomalies in buccal smears: Methods development. Mutat. Res. 1992, 271, 69–77. [Google Scholar] [CrossRef]
  28. Thomas, P.; Holland, N.; Bolognesi, C.; Kirsch-Volders, M.; Bonassi, S.; Zieger, E.; Knasmueller, S.; Fenech, M. Buccal micronucleus cytome assay. Nat. Protoc. 2009, 4, 825–837. [Google Scholar] [CrossRef] [PubMed]
  29. Biesterfeld, S.; Beckers, S.; Del Carmen Villa Cadenas, M.; Schramm, M. Feulgen staining remains the gold standard for precise DNA image cytometry. Anticancer Res. 2011, 31, 53–58. [Google Scholar] [PubMed]
  30. Bolognesi, C.; Fenech, M. Micronucleus Cytome Assays in Human Lymphocytes and Buccal Cells. Methods. Mol. Biol. 2019, 2031, 147–163. [Google Scholar] [CrossRef]
  31. Holland, N.; Bolognesi, C.; Kirsch-Volders, M.; Bonassi, S.; Zeiger, E.; Knasmueller, S.; Fenech, M. The micronucleus in human buccal cells as a tool for biomonitoring DNA damage: The HUMN project perspective on current status and knowledge gaps. Mutat. Res. 2008, 659, 93–108. [Google Scholar] [CrossRef]
  32. Damián-Huato, M.A.; Ramírez-Valverde, B. Dependencia científica y tecnologías campesinas. El caso de los productores de maíz del estado de Tlaxcala. Econ. Soc. 2008, 14, 59–76. [Google Scholar]
  33. Carbajal-López, Y.; Gómez-Arroyo, S.; Villalobos-Pietrini, R.; Calderón-Segura, M.; Martínez-Arroyo, A. Biomonitoring of agricultural workers exposed to pesticide mixtures in Guerrero state, Mexico, with comet assay and micronucleus test. Environ. Sci. Pollut. Res. 2016, 23, 2513–2520. [Google Scholar] [CrossRef]
  34. Ramírez-Mora, E.; Pérez-Vázquez, A.; Landeros-Sánchez, C.; Martínez-Dávila, J.; Villanueva-Jimenez, J.; Lagunés-Espinoza, L.C. Occupational exposure to pesticides in sugarcane agroecosystems in the central region of Veracruz state, Mexico. Rev. Bio Cienc. 2019, 6, 495. [Google Scholar] [CrossRef]
  35. Damián-Huato, M.A. Manejo del maíz en el estado de Tlaxcala, México: Entre lo convencional y lo agroecológico. Rev. Lat. Rec. Nat. 2010, 6, 67–76. [Google Scholar]
  36. Nicholson, C.C.; Williams, N.M. Cropland heterogeneity drives frequency and intensity of pesticide use. Environ. Res. Lett. 2021, 16, e074008. [Google Scholar] [CrossRef]
  37. Nboyine, J.A.; Asamani, E.; Agboyi, L.K. Assessment of the optimal frequency of insecticide sprays required to manage fall armyworm (Spodoptera frugiperda J.E Smith) in maize (Zea mays L.) in northern Ghana. CABI Agric. Biosci. 2022, 3, e3. [Google Scholar] [CrossRef]
  38. Hutter, H.-P.; Poteser, M.; Lemmerer, K.; Wallner, P.; Shahraki Sanavi, S.; Kundi, M.; Moshammer, H.; Weitensfelder, L. Indicators of Genotoxicity in Farmers and Laborers of Ecological and Conventional Banana Plantations in Ecuador. Int. J. Environ. Res. Public Health 2020, 17, 1435. [Google Scholar] [CrossRef] [PubMed]
  39. Bolognesi, C.; Fenech, M. Micronucleus assay in human cells: Lymphocytes and buccal cells. Methods Mol. Biol. 2013, 1044, 191–207. [Google Scholar] [CrossRef] [PubMed]
  40. Bloching, M.; Hofmann, A.; Lautenschläger, C.; Berghaus, A.; Grummt, T. Exfoliative cytology of normal buccal mucosa to predict the relative risk of cancer in the upper aerodigestive tract using the MN-assay. Oral Oncol. 2000, 36, 550–555. [Google Scholar] [CrossRef]
  41. Arévalo-Jaramillo, P.; Idrobo, A.; Salcedo, L.; Cabrera, A.; Vintimilla, A.; Carrión, M.; Bailon-Moscoso, N. Biochemical and genotoxic effects in women exposed to pesticides in Southern Ecuador. Environ. Sci. Pollut. Res. Int. 2019, 26, 24911–24921. [Google Scholar] [CrossRef] [PubMed]
  42. Hutter, H.-P.; Khan, A.W.; Lemmerer, K.; Wallner, P.; Kundi, M.; Moshammer, H. Cytotoxic and Genotoxic Effects of Pesticide Exposure in Male Coffee Farmworkers of the Jarabacoa Region, Dominican Republic. Int. J. Environ. Salud Pública 2018, 15, 1641. [Google Scholar] [CrossRef]
  43. Salvi, R.M.; Lara, D.R.; Ghisolfi, E.S.; Portela, L.V.; Dias, R.D.; Souza, D.O. Neuropsychiatric evaluation in subjects chronically exposed to organophosphate pesticides. Toxicol. Sci. 2003, 72, 267–271. [Google Scholar] [CrossRef]
  44. Claudio, S.R.; Simas, J.M.M.; Souza, A.C.F.; De Alencar, M.D.C.B.; Yamauchi, L.Y.; Ribeiro, D.A. Genomic Instability and Cytotoxicity in Buccal Mucosal Cells of Workers in Banana Farming Evaluated by Micronucleus Test. Anticancer Res. 2019, 39, 1283–1286. [Google Scholar] [CrossRef]
  45. Benedetti, D.; Nunes, E.; Sarmento, M.; Porto, C.; Santos, C.E.I.; Dias, J.F.; da Silva, J. Genetic damage in soybean workers exposed to pesticides: Evaluation with the comet and buccal micronucleus cytome assays. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2013, 752, 28–33. [Google Scholar] [CrossRef]
  46. Dutta, S.; Bahadur, M. Cytogenetic analysis of micronuclei and cell death parameters in epithelial cells of pesticide exposed tea garden workers. Toxicol. Mech. Methods. 2016, 26, 627–634. [Google Scholar] [CrossRef]
  47. Ferré, D.M.; Quero, M.; Hynes, V.; Saldeña, E.; Lentini, V.; Tornello, M.; Carracedo, R.; Gorla, N.B. Ensayo de micronúcleos de citoma bucal en trabajadores de fincas frutícola que han aplicado plaguicidas alrededor de quince años. Rev. Int. Contam. 2018, 34, 23–33. [Google Scholar] [CrossRef]
  48. Martínez-Valenzuela, C.; Waliszewski, S.M.; Amador-Muñoz, O.; Meza, E.; Calderón-Segura, M.E.; Zenteno, E.; Huichapan-Martínez, J.; Caba, M.; Félix-Gastélum, R.; Longoria-Espinoza, R. Aerial pesticide application causes DNA damage in pilots from Sinaloa, Mexico. Environ. Sci. Pollut. Res. 2017, 24, 2412–2420. [Google Scholar] [CrossRef] [PubMed]
  49. Ortega-Martínez, L.D.; Pérez-Armendáriz, B.; Waliszewski, S.; Gómez-Arroyo, S.; Baños-Lara, M.R.; Terán-Cervantes, M.; Castro-Carranza, G.; Martínez-Valenzuela, M.C. Daño genético y citotóxico provocado por plaguicidas en invernaderos en Atlixco, Puebla, México. Rev. Int. Contam. 2019, 35, 271–279. [Google Scholar] [CrossRef]
  50. Gentile, N.; Mañas, F.; Bosch, B.; Peralta, L.; Gorla, N.; Aiassa, D. Micronucleus Assay as a Biomarker of Genotoxicity in the Occupational Exposure to Agrochemicals in Rural Workers. Bull. Environ. Contam. Toxicol. 2012, 88, 816–822. [Google Scholar] [CrossRef] [PubMed]
  51. Sharma, A.; Kumar, V.; Shahzad, B. Worldwide pesticide usage and its impacts on ecosystem. SN Appl. Sci. 2019, 1, e1446. [Google Scholar] [CrossRef]
  52. Parra-Arroyo, L.; Gonzalez-Gonzalez, R.B.; Castillo-Zacarias, C.; Melchor Martinez, E.M.; Sosa-Hernandez, J.E.; Bilal, M.; Iqbal, H.M.N.; Barcelo, D.; Parra-Saldivar, R. Highly hazardous pesticides, and related pollutants: Toxicological, regulatory, and analytical aspects. Sci. Total Environ. 2022, 807, 151879. [Google Scholar] [CrossRef]
  53. Ramírez, A.M.; Haller, V.V.; Romero, M.L.G. Sistemas de producción agrícola de maíz en tres municipios de Tlaxcala: Los componentes tecnológicos de la Revolución Verde con relación a la tecnología tradicional. Reg. Y Desarro. Sustentable 2019, 19, 67–93. [Google Scholar]
  54. Aiassa, D.E.; Mañas, F.J.; Gentile, N.E.; Bosch, B.; Salinero, M.C.; Gorla, N.B.M. Evaluation of genetic damage in pesticides applicators from the province of Córdoba, Argentina. Environ. Sci. Pollut. Res. 2019, 26, 20981–20988. [Google Scholar] [CrossRef]
  55. World Health Organization (WHO). Recommended Classification of Pesticides by Hazard and Guidelines to Classification 2009. Available online: https://www.who.int/iFCs/publications/pesticides_hazard_2009.pdf (accessed on 2 October 2021).
  56. Ferraz, G.A.; Costa Neto, A.O.; Cerqueira, E.M.M.; Meireles, J.R.C. Effects of age on the frequency of micronuclei and degenerative 105 nuclear abnormalities. Rev. Bras. Geriatr. Gerontol. 2016, 19, 627–634. [Google Scholar] [CrossRef]
  57. Vindas, R.; Ortiz, F.; Ramírez, V.; Cuenca, P. Genotoxicity of three pesticides used in Costa Rican banana plantations. Rev. Biol. Trop. 2004, 52, 601–609. [Google Scholar]
  58. Matheus, T.; Aular, Y.; Bolaños, A.; Fernández, Y.; Barrios, E.; Hung, M.-L. Actividad de butirilcolinesterasa y micronúcleos en trabajadores agrícolas expuestos a mezclas de plaguicidas. Salud Trab. 2017, 25, 23–36. [Google Scholar]
  59. Larrea-Poma, M.; Tirado-Bustillos, N.; Ascarrunz, G.M.E. Daño genotóxico por exposición a plaguicidas en agricultores del Municipio de Luribay. Biofarbo 2010, 18, 31–43. [Google Scholar]
  60. Tomiazzi, J.S.; Judai, M.A.; Nai, G.A.; Pereira, D.R.; Antunes, P.A.; Favareto, A.P.A. Evaluation of genotoxic effects in Brazilian agricultural workers exposed to pesticides and cigarette smoke using machine-learning algorithms. Environ. Sci. Pollut. Res. 2018, 25, 1259–1269. [Google Scholar] [CrossRef] [PubMed]
  61. Damalas, C.A.; Koutroubas, S.D. Farmers’ Exposure to Pesticides: Toxicity Types and Ways of Prevention. Toxics 2016, 4, 1. [Google Scholar] [CrossRef] [PubMed]
  62. Lesmes-Fabian, C.; García-Santos, G.; Leuenberger, F.; Nuyttens, D.; Binder, C.R. Dermal exposure assessment of pesticide use: The case of sprayers in potato farms in the Colombian highlands. Sci. Total Environ. 2012, 430, 202–208. [Google Scholar] [CrossRef]
  63. Moshou, H.; Karakitsou, A.; Yfanti, F.; Hela, D.; Vlastos, D.; Paschalidou, A.K.; Kassomenos, P.; Petrou, I. Assessment of genetic effects and pesticide exposure of farmers in NW Greece. Environ. Res. 2020, 186, e109558. [Google Scholar] [CrossRef]
  64. Joko, T.; Dewanti, N.A.Y.; Dangiran, H.L. Pesticide Poisoning and the Use of Personal Protective Equipment (PPE) in Indonesian Farmers. J. Environ. Public Health. 2020, 1–7. [Google Scholar] [CrossRef] [Green Version]
  65. Guzmán-Plazola, P.; Guevara-Gutiérrez, R.D.; Olguín-López, J.L.; Mancilla-Villa, O.R. Perspectiva campesina, intoxicaciones por plaguicidas y uso de agroquímicos. Idesia 2016, 34, 69–80. [Google Scholar] [CrossRef] [Green Version]
Applsci 12 09050 g001 550
Figure 1. Overview of the geographical location of the study site (locality of Vicente Guerrero, Españita-Tlaxcala, Mexico) [21].
Figure 1. Overview of the geographical location of the study site (locality of Vicente Guerrero, Españita-Tlaxcala, Mexico) [21].
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Figure 2. Mean frequency (95% confidence intervals) of nuclear anormalities per 1000 cells in each pool (FC = group of conventional farmers; FA = group of farmers using agroecological practices): (a) total number of micronuclei (MNi); (b) condensed chromatin (CC); (c) karyolysis (KL); (d) binucleate cell (BN); (e) micronuclei (MN); (f) karyorrhexis (KR); (g) pyknosis (PY); (h) basal cells (BC); (i) lobed nuclei (BUD). * Significant statistical differences. *** Highly significant statistical differences.
Figure 2. Mean frequency (95% confidence intervals) of nuclear anormalities per 1000 cells in each pool (FC = group of conventional farmers; FA = group of farmers using agroecological practices): (a) total number of micronuclei (MNi); (b) condensed chromatin (CC); (c) karyolysis (KL); (d) binucleate cell (BN); (e) micronuclei (MN); (f) karyorrhexis (KR); (g) pyknosis (PY); (h) basal cells (BC); (i) lobed nuclei (BUD). * Significant statistical differences. *** Highly significant statistical differences.
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Figure 3. Cells with degenerative nuclear changes related to genotoxic damage of the FC group: (a) normal cells; (b) condensed chromatin (CC); (c) karyolysis (KL); (d) karyorrhexis (KR); (e) pyknotic nuclei (PY); (f) binucleated cells (BN); (g) lobulated nuclei (BUD); (h,i) cells with micronuclei (MN).
Figure 3. Cells with degenerative nuclear changes related to genotoxic damage of the FC group: (a) normal cells; (b) condensed chromatin (CC); (c) karyolysis (KL); (d) karyorrhexis (KR); (e) pyknotic nuclei (PY); (f) binucleated cells (BN); (g) lobulated nuclei (BUD); (h,i) cells with micronuclei (MN).
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Table
Table 1. Sociodemographic characteristics of the groups under study.
Table 1. Sociodemographic characteristics of the groups under study.
VariablesGroup with the Highest Use of PesticidesControl Group, with Less Use of Pesticides
FC = 47FA = 43
FrequencyPercentage (%)FrequencyPercentage (%)
Age<46 years1327.66920.93
47–59 years2144.681944.19
60–72 years1123.401227.91
73–84 years24.2636.98
No education48.5124.65
Level of educationElementary education2757.451637.21
Secondary education1429.792251.16
Technical education24.2636.98
Smoking habitNo smoking3778.723683.72
1–5 cigarettes/day24.2649.30
6–10 cigarettes/day36.3824.65
More than 10 cigarettes/day510.6412.33
Alcohol consumptionNo alcohol24.2612.33
Once a week1123.401227.91
2 to 3 times a week2859.572660.47
4 or more times a week612.7749.30
FC = group of conventional farmers; FA = group of agroecological farmers.
Table
Table 2. Pesticides used by peasant maize producers in the municipality of Vicente Guerrero, Tlaxcala, Mexico.
Table 2. Pesticides used by peasant maize producers in the municipality of Vicente Guerrero, Tlaxcala, Mexico.
Ingredient ActiveChemical FamilyCommercial NameType 1TC 2List PAP 3Type of Pesticides Used
Group with the Highest Use of Pesticides (FC)Control Group, with Less Use of Pesticides (FA)
TriasulfuronSulfonylureaAmber 75 GSHIII-19.14% (9/47)4.65% (2/43)
Atrazine + MesotrioneTriazine + TriketoneCallisto XtraHIII-4.25% (2/47)4.65% (2/43)
DicambaBenzoic acids2-CambaHII-8.51% (4/47)2.32% (1/43)
CypermethrinPyrethroidsArrivo 200,
Combat 20,
Fipol 200		III331.8% (14/47)-
2,4-DClorofenoxiHerbipo l 4,
Hierbamina,
Esteron 47		HII-40.42% (19/47)18.60% (8/43)
GlyphosateOrganophosphate (Glycine)FaenaHIII18.51% (4/47)9.30% (4/43)
AtrazineTriazineGesaprimHIII240.42% (19/47)13.95% (6/43)
NicosulfuronSulfonylureaSanson 4SCHU-12.76% (6/47)4.65% (2/43)
ClethodimCyclohexanedionesSelect ultraHIII-10.63% (5/47)2.32% (1/43)
CarbofuranCarbamateCurater 4f,
Furadan 350		I, NIb1,3,42.12% (1/47)-
Lambda cyalotrinPyrethroidsKarate Zeon 5III1,2,331.8% (14/47)-
MalathionOrganophosphateMalathion 1000IIII2,340.42% (19/47)-
TebuconazoleBenzimidazoleFolicur 250FII-23.40% (11/47)-
MancozebDithiocarbamatesManzate superFU219.14% (9/47)-
AzoxystrobinMethoxyacrylatesMaxidor 250FU-12.76% (6/47)-
Hydrogen peroxide + Peroxyacetic acidPeroxyacidOxicureF, NII-4.25% (2/47)-
FC = group of conventional farmers; FA = group of agroecological practices farmers. (1) Type: I = insecticide; H = herbicide; F = fungicide; N = nematicide. (2) Toxicological category (TC): Ib = very hazardous; II = moderately hazardous; III = slightly hazardous; U = unlikely to present an acute hazard. (3) Criteria for inclusion in the list of highly hazardous pesticides (HHP): 1 = High acute toxicity; 2 = chronic effects on human health; 3 = environmental toxicity; 4 = restricted or banned by environmental conventions [25].
Table
Table 3. Total micronuclei (MNi) evaluated in peasant corn producers according to the sociodemographic characteristics of the locality of Vicente Guerrero in the municipality of Españita, Tlaxcala, Mexico.
Table 3. Total micronuclei (MNi) evaluated in peasant corn producers according to the sociodemographic characteristics of the locality of Vicente Guerrero in the municipality of Españita, Tlaxcala, Mexico.
VariablesN%Group with the Highest Use of Pesticides (FC)Control Group, with Less Use of Pesticides (FA)p Value
nMNi%nMNi%
Total MNi90100470.40852.2430.25347.80.001 *
Age<46 years2224.4130.41914.490.237100.021 *
47–59 years4044.4210.42123.3190.25021.1
60–72 years2425.5110.36712.2120.25713.3
73–84 years55.620.4302.230.3073.4
Level of educationNo education66.640.4534.420.4602.20.02 *
Elementary education4347.8270.41330160.31717.8
Secondary education3640140.39315.6220.19124.4
Technical education55.520.3602.230.2303.3
Smoking habitNo smoking7280370.38541.1360.28138.90.02 *
1–5 cigarettes/day86.620.4852.240.0804.4
6–10 cigarettes/day55.530.5073.320.2702.2
Más de 10 cigarettes/day67.850.4945.610.092.2
Alcohol consumptionNo alcohol33.320.5402.210.1101.10.006 *
Once a week2325.5110.37612.2120.23613.3
2 to 3 times a week5460280.42831.1260.25228.9
4 or more times a week1011.160.3306.740.3434.4
Personal 1
Protective equipment		Does not use3741.1250.49827.8120.478813.30.001 *
1 to 2 protective equipment used1415.690.3761050.3505.6
2 to 3 protective equipment used1415.680.3008.960.2836.7
3 to 5 protective equipment used2527.850.1895.6200.08422.2
FC = group of conventional farmers; FA = group of agroecological farmers; N = total population of corn producers; n = population per group of corn producers; % = percentage; MNi = total number of micronuclei. 1 Protective equipment used: (a) use of long sleeves, (b) use of protective glasses, (c) use of nasal mask, (d) use of gloves, and (e) use of boots. * Significant statistical differences (p ≤ 0.05).
Table
Table 4. Comparison of BMCA results between exposed (FC) and control (FA) groups.
Table 4. Comparison of BMCA results between exposed (FC) and control (FA) groups.
VariablesMedianIQRMann–Whitney
U Test		Z1-βdSignificance
p ≤ 0.05
MNi(FC) = 0.3800.180533.50−3.850.990.930.001 ***
(FA) = 0.2700.300
CC(FC) = 0.6700.328547.50−3.740.910.720.001 ***
(FA) = 0.4130.238
KL(FC) = 0.2220.225929.50−0.650.050.040.513
(FA) = 0.1390.165
BN(FC) = 0.0000.000991.50−3.180.070.100.612
(FA) = 0.0000.000
MN(FC) = 0.0070.009389.00−5.040.991.080.001 ***
(FA) = 0.0030.003
KR(FC) = 0.0220.360516.50−3.990.950.790.001 ***
(FA) = 0.0070.110
PY(FC) = 0.0360.360704.00−2.480.800.610.013 *
(FA) = 0.0240.170
BC(FC) = 0.0000.00908.50−1.390.270.290.162
(FA) = 0.0000.00
BUD(FC) = 0.0000.000795.00−3.180.820.550.001 ***
(FA) = 0.0000.000
FC = group of conventional farmers; FA = group of agroecological farmers; BMCA = buccal micronucleus cytome assay. Note: Total number of micronuclei (MNi), condensed chromatin (CC), karyolysis (KL), binucleate cells (BN), micronuclei (MN), karyorrhexis (KR), pyknosis (PY), basal cells (BC), lobed nucleus (BUD). IQR = interquartile range; 1-β = power of a study; d = effect size d. * Significant statistical differences. *** Highly significant statistical differences according to Mann–Whitney U test (p ≤ 0.05).
Table
Table 5. Total micronuclei (MNi) with respect to sociodemographic variables, confounding factors, and personal protection measures of corn producers from the town of Vicente Guerrero in the municipality of Españita, Tlaxcala, Mexico.
Table 5. Total micronuclei (MNi) with respect to sociodemographic variables, confounding factors, and personal protection measures of corn producers from the town of Vicente Guerrero in the municipality of Españita, Tlaxcala, Mexico.
Groups/Results VariablesMNiAdjusted Odds Ratio [AOR] †(95% CI) †p Interact †,*
Group FC47 (52.2)1.6611.306–2.112≤0.001 *
Group FA43 (47.8)1REF
Age
Groups/Results variables<46
years		47–59
years		60–72
years		73–84
years		Adjusted Odds Ratio [AOR] †(95% CI) †p interact †,*
Group FC13 (27.70)21 (44.70)11 (23.40)2 (4.30)0.9930.971–1.0150.759
Group FA9 (20.90)19 (44.20)12 (27.90)3 (7.00)1REF
Leve of education
Groups/Results variablesNo
education		Elementary
education		Secondary
education		Technical
education		Adjusted Odds Ratio [AOR] †(95% CI) †p interact †,*
Group FC4 (8.50)27 (57.40)14 (29.80)2 (94.30)0.9860.959–1.0130.659
Group FA2 (4.70)16 (37.20)22 (51.20)3 (7.00)1REF
Smoking habit
Groups/Results variablesNo
smoking		1–5
cigarettes/day		6–10
cigarettes/day		>10
cigarettes/day		Adjusted Odds Ratio [AOR] †(95% CI) †p interact †,*
Group FC37 (78.70)2 (4.30)3 (6.40)5 (10.60)1.0080.988–1.0290.610
Group FA35 8(1.40)4 (9.30)2 (4.70)2 (4.70)1REF
Alcohol consumption
Groups/Results variablesNo
alcohol		Once
a week		2 to 3
times a week		4 or more times a weekAdjusted Odds Ratio [AOR] †(95% CI) †p interact †,*
Group FC2 (4.30)11 (23.90)28 (59.60)6 (12.80)0.9960.969–1.0240.893
Group FA1 (2.30)12 (27.90)26 (60.50)4 (9.30)1REF
1 Personal protective equipment
Groups/Results variablesDoes
not use		1 to 2
protective equipment used		2 to 3
protective equipment used		3 to 5
protective equipment used		Adjusted Odds Ratio [AOR] †(95% CI) †p interact †,*
Group FC25 (40.10)9 (29.10)8 (19.20)5 (11.60)0.8840.874–0.903≤0.003 *
Group FA12 (39.20)5 3 (32.60)6 (17.80)20 (10.40)1REF
FC = group of conventional farmers; FA = group of agroecological farmers; MNi = total number of micronuclei. † Adjusted for producer group, age, level of education, smoking habit, alcohol consumption, and personal protective equipment. Numbers in parentheses indicate the percentage strengths of associations with respect to the respondents in each category of the outcome variables. 1 Protective equipment used: (a) use of long sleeves, (b) use of protective glasses, (c) use of nasal mask, (d) use of gloves, (e) use of boots. Note: * p values for interactions from the likelihood ratio test.
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Rivera, A.; Cedillo Ramírez, L.; Parraguirre Lezama, C.; Baez Simon, A.; Laug Garcia, B.; Romero-Arenas, O. Evaluation of Cytotoxic and Genotoxic Risk Derived from Exposure to Pesticides in Corn Producers in Tlaxcala, Mexico. Appl. Sci. 2022, 12, 9050. https://doi.org/10.3390/app12189050

AMA Style

Rivera A, Cedillo Ramírez L, Parraguirre Lezama C, Baez Simon A, Laug Garcia B, Romero-Arenas O. Evaluation of Cytotoxic and Genotoxic Risk Derived from Exposure to Pesticides in Corn Producers in Tlaxcala, Mexico. Applied Sciences. 2022; 12(18):9050. https://doi.org/10.3390/app12189050

Chicago/Turabian Style

Rivera, Antonio, Lilia Cedillo Ramírez, Conrado Parraguirre Lezama, Alfredo Baez Simon, Beatriz Laug Garcia, and Omar Romero-Arenas. 2022. "Evaluation of Cytotoxic and Genotoxic Risk Derived from Exposure to Pesticides in Corn Producers in Tlaxcala, Mexico" Applied Sciences 12, no. 18: 9050. https://doi.org/10.3390/app12189050

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