19 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 Luciane Baierle Lorenzatto Mestra em Ciências da Saúde pela Universidade Comunitária da Região de Chapecó (Unochapecó), Santa Catarina, Brasil. Maria Isabel Gonçalves da Silva Mestra em Ciências da Saúde pela Unochapecó – Chapecó (SC), Brasil. Walter Antonio Roman Junior Docente do Programa de Pós- Graduação Stricto Sensu em Ciências da Saúde da Unochapecó – Chapecó (SC), Brasil. Sinval Adalberto Rodrigues Junior Docente do Programa de Pós-Graduação Stricto Sensu em Ciências da Saúde da Unochapecó – Chapecó (SC), Brasil. Clodoaldo Antônio De Sá Docente e coordenador do Programa de Pós-Graduação Stricto Sensu em Ciências da Saúde da Unochapecó – Chapecó (SC), Brasil. Vanessa da Silva Corralo Docente do Programa de Pós-Graduação Stricto Sensu em Ciências da Saúde da Unochapecó – Chapecó (SC), Brasil. Endereço para correspondência: Vanessa da Silva Corralo – Servidão Anjo da Guarda, 295-D, Efapi – CEP 89809-900 – Chapecó (SC), Brasil – E-mail: vcorralo@unochapeco.edu.br Recebido em: 12/09/2019 Aceito em: 13/02/2020 ABSTRACT Pesticides are compounds widely used in agriculture, due the productive efficiency increase. However, these toxic compounds can exert negative effects to environment and human health. This work aim was analyzing the occupational exposure of rural workers to pesticides, through a descriptive-analytical study with cross-sectional and quantitative approach. The sample consisted of 63 farm workers. Socioeconomic profile characterization, pesticide management, and health conditions was carried out through interviews. After biological samples collection, erythrocyte acetylcholinesterase enzyme activity was assessed in two moments: pre- and post-exposure to pesticides in one of planting and harvesting cycle. Rural workers age mean was 48.1 ± 7.6 years. They reported using personal protective equipment, although incompletely or inadequately, and 46% reported pesticide poisoning. Mood disorders such as anxiety (50.3%) and depressive state (27.0%) were also reported by workers. There was a significant decrease in the activity of the erythrocyte acetylcholinesterase enzyme from 0.83 ± 0.06 delta pH/hour to 0.71 ± 0.11 delta pH/hour (p < 0.05). Individual comparison of enzyme activity values showed that 19.6% of the workers were intoxicated with carbamate and organophosphate. Contamination is attributed to the inadequate management of pesticides. Adopting measures to minimize intoxications and other harmful effects on the health of rural workers is considered urgent. Keywords: agrochemicals; occupational exposure; biomarkers; acetylcholinesterase. RESUMO Os pesticidas são compostos amplamente utilizados na agricultura, por causa do aumento da eficiência produtiva, no entanto esses compostos tóxicos podem exercer efeitos negativos ao meio ambiente e à saúde humana. O objetivo deste trabalho foi analisar a exposição ocupacional de trabalhadores rurais a agrotóxicos, através de um estudo descritivo-analítico com abordagem transversal e quantitativa. A amostra foi composta de 63 trabalhadores agrícolas. A caracterização do perfil socioeconômico, o manejo de pesticidas e as condições de saúde foram realizados por meio de entrevistas. Após a coleta das amostras biológicas, a atividade da enzima acetilcolinesterase eritrocitária foi avaliada em dois momentos: pré e pós-exposição a pesticidas em um ciclo de plantio e colheita. A idade média dos trabalhadores rurais foi de 48,1 ± 7,6 anos. Eles relataram usar equipamentos de proteção individual, embora de forma incompleta ou inadequadamente, e 46% relataram intoxicação por agrotóxicos. Transtornos do humor como ansiedade (50,3%) e estado depressivo (27,0%) também foram relatados pelos trabalhadores. Houve uma diminuição significativa na atividade da enzima acetilcolinesterase eritrocitária de 0,83 ± 0,06 delta pH/hora para 0,71 ± 0,11 delta pH/hora (p < 0,05). A comparação individual dos valores da atividade enzimática mostrou que 19,6% dos trabalhadores estavam intoxicados com carbamatos e organofosforados. A contaminação é atribuída ao manejo inadequado de pesticidas. Considera-se urgente a adoção de medidas para minimizar intoxicações e outros efeitos nocivos à saúde dos trabalhadores rurais. Palavras-chave: agrotóxicos; exposição ocupacional; biomarcadores; acetilcolinesterase. DOI: 10.5327/Z2176-947820200528 RURAL WORKERS EXPOSURE TO ORGANOPHOSPHATES AND CARBAMATES EXPOSIÇÃO DE TRABALHADORES RURAIS A ORGANOFOSFORADOS E CARBAMATOS http://n.XX https://orcid.org/0000-0002-8118-8206 https://orcid.org/0000-0001-7768-7531 https://orcid.org/0000-0001-8363-8795 https://orcid.org/0000-0002-4475-1725 https://orcid.org/0000-0001-7409-8870 https://orcid.org/0000-0003-4234-4875 mailto:vcorralo@unochapeco.edu.br Lorenzatto, L.B. et al. 20 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 INTRODUCTION With the changes in the agricultural production pro- cess that have occurred in recent decades, espe- cially with regard to the intensive use of pesticides, the damage caused to the environment has become increasingly evident (LOPES; ALBUQUERQUE, 2018; MAJOLO; REMPEL, 2018). As pesticides can contam- inate water, air, and soil, the natural habitat of nu- merous species becomes altered, causing significant negative impacts on ecosystems. Fish, insects, am- phibians, among other animals, suffer from poisoning by pesticides present in the environment (CHELINHO et al., 2012; GONÇALVES et al., 2015). Similar to what happens with different species of an- imals in nature, the use of pesticides has affected hu- man health, increasing the risk of diseases and work accidents related to intoxication, especially among rural workers (PIGNATI et al., 2017). Rural work- ers represent the population group most exposed to pesticides and, consequently, most vulnerable to health problems resulting from poisoning with these compounds (MURUSSI et al., 2014; STRELITZ; ENGEL; KEIFER, 2014). Among the health problems, the ad- verse effects on the central nervous system (CNS) and peripheral system, increased incidence of cancer, malformations, and endocrine and immune disorders can be mentioned (JONAS, 2015; KIM; KABIR; JAHAN, 2017; MOSTAFALOU; ABDOLLAHI, 2017). The greater susceptibility of this population group is related to factors such as neglect in the use of personal protec- tive equipment (PPE), since rural workers do not use it, or use equipment that is not appropriate, making them even more vulnerable to exposure to pesticides (SIQUEIRA et al., 2013). Organophosphates and carbamates are among the most used pesticides and the most frequently asso- ciated with occupational poisoning (LOZANO-PANI- AGUA et al., 2016). These substances act as inhibitors of the enzyme acetylcholinesterase (AChE), causing an accumulation of acetylcholine at cholinergic recep- tors. Thus, postsynaptic cholinergic transmission does not cease in adequate time, leading to hyperstimula- tion, which may trigger symptoms such as headache, tremors, dizziness, loss of consciousness and fainting (EDDLESTON et al., 2012; ANDRADE FILHO; SOUZA, 2013; SIQUEIRA et al., 2013). The level of acetylcholinesterase enzyme activity in the blood becomes an important biomarker to demon- strate the exposure to pesticides of the organophos- phate and carbamate classes, still used despite their harmful effects on health (MURUSSI et al., 2014; MAGNARELLI, 2015). In this sense, the objective of this study was to analyze occupational exposure to pesti- cides and the relationship between exposure and lev- els of the enzyme acetylcholinesterase in rural workers in western Santa Catarina. MATERIALS AND METHODS This is a descriptive-analytical study with a cross-sectional and quantitative approach involving rural workers oc- cupationally exposed to pesticides. Population, sample, and selection The study population consisted of 658 male rural workers from a rural municipality in western San- ta Catarina, aged between 19 and 60 years (SISAB, 2017). The G* Power 3.1 software was used to cal- culate the sample size. A significance level of 5%, test power of 80.0%, equal standard deviations in pre- and post-exposure scores, and a correlation co- efficient of 0.30 (minimum value to be detected in the consistency assessment) were considered. Based on this calculation, although the minimum sample size required was 35 participants, 63 rural workers were included. For the constitution of the sample, the subjects were drawn from random numbers gen- erated on a website generating random numbers. Each participant drawn to compose the sample was visited at home by a researcher properly trained to perform data collection. http://n.XX Rural workers exposure to organophosphates and carbamates 21 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 Ethical aspects This study followed resolution 466 / CNS / 2012 (BRA- SIL, 2012), of the National Health Council of Brazil, which establishes guidelines to preserve the dignity, rights, security, and well-being of research partici- pants. This study was approved by the Research Eth- ics Committee Involving Human Beings of Unochapecó University (protocol number 2100319). All subjects in the sample were informed about the ob- jectives and procedures of the study and signed an In- formed Consent Form, in two copies, authorizing their participation in the research. A copy of the term was given to the participant and another was under the responsibility of the researchers, together with all the data collected in the study. Interview and data collection instrument Initially, rural workers answered a questionnaire ap- plied in the form of interviews, with questions relat- ed to socioeconomic and demographic characteristics, knowledge about the use of pesticides, and health con- ditions. The application of the questionnaire occurred in an individualized way at the worker’s home. The socioeconomic profile was evaluated through the application of the questionnaire based on the Brazilian Economic Classification Criteria of the Brazilian Asso- ciation of Research Companies (Associação Brasileira de Empresas de Pesquisa — ABEP). These criteria are based on an economic classification tool that uses a survey of the characteristics of the household (pres- ence and quantity of some domestic comfort items and level of education of the head of the family) to differentiate the population (ABEP, 2015). Each item of the instrument is scored and, at the end, the results are added up and the individual’s socioeconomic clas- sification is performed, observing the following cutoff points: class A: 45-100 points; class B1: 38-44 points; class B2: 29-37 points; class C1: 23-28 points; class C2: 17-22 points; and classes D-E: 0-16 points (ABEP, 2015). During data collection, collections of blood samples were scheduled for further analysis of acetylcholines- terase activity. Enzyme dosage To determine exposure to carbamate and organophos- phate classes pesticides by measuring the enzymatic activity of erythrocyte acetylcholinesterase, two 4-mL blood samples were collected in EDTA tubes at the out- patient clinic of the municipality’s Health Unit, in two moments. The first collection of blood samples oc- curred before the agricultural planting period (pre-ex- posure) and the second collection occurred after five months, at the end of the harvest (post-exposure). AChE enzyme activity was analyzed by the potentio- metric method, which allows quantifying from elec- trodes, the pH variation of a medium containing the sample (erythrocyte solution) and the acetylcholine substrate. The enzyme acetylcholinesterase present in erythrocytes should hydrolyze acetylcholine and, as a consequence, release acetic acid. The acidification of the medium promotes quantifiable pH variation, which can be measured by an acid-sensitive electrode, and the enzymatic activity can be expressed as pH variation per hour (delta pH/h). The normal reference values range from 0.58 to 0.95 delta pH/h for men (MI- CHEL, 1949; HOLAS et al., 2012). According to Ribeiro and Mella (2007), it is necessary to perform the pre-exposure baseline test in workers who will have contact with organophosphates and car- bamates, considering that the cholinesterase basal lev- els vary from one person to another. Statistical analysis Data were presented as means and respective stan- dard deviations or absolute and relative frequencies (%). The normality of the continuous data was analyzed by the Shapiro-Wilk test. The Wilcoxon test for related samples was used to compare pre- and post-exposure data. The Statistical Package for the Social Sciences (SPSS®), version 20.0 was used for all analyses. The sig- nificance level was set at 5% (p < 0.05). http://n.XX Lorenzatto, L.B. et al. 22 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 RESULTS AND DISCUSSION Characterization of the study population and contact with pesticides The study sample consisted of 63 rural male workers, with a mean age of 48.1 ± 7.6 years, of whom 92.1% were married. Regarding education, 39.7% had ele- mentary education and 28.6% had primary education. In general, 68.3% of the subjects in the sample studied for up to eight years. Most workers were classified in the middle and low economic classes, with 57.1% in class B (B1 and B2) and 38.0% in class C (C1 and C2). The average pesticide exposure time for rural workers in the present study was 24.8 ± 8.3 years. Although 95.2% of individuals reported using PPE, these were not always adequate or sufficient to protect them from pesticides (Table 1). Only 39.7% used the mask with a filter that provides effective protection, 79.0% stat- ed they used gloves, 20.6% used goggles, and 88.9% mentioned the use of coveralls (Table 1). These data demonstrate a high degree of vulnerability of these workers due to the conditions under which they are exposed to pesticides, a condition also observed in oth- er studies (SIQUEIRA et al., 2013; WAHLBRINCK; BICA; REMPEL, 2017; CORCINO et al., 2019). The workers reported that the non-use of PPE is relat- ed to the fact that some equipment causes thermal discomfort, making it difficult to perform the tasks. For this reason, especially in small rural communities, it is common to find rural workers without PPE during the handling and application of pesticides, despite the mandatory use of such equipment. It is important to consider that washing the PPE is also an important source of contamination because the PPE is washed and stored by other people in the family, usually the wife. Another worrying aspect is the wash- ing of clothes used during the application of pesticides, since 41.3% of workers stated that their clothes were washed together with those of the family (GONÇALVES; MELO, 2014). Farmworkers were also asked about direct skin contact with pesticides, and 66.7% admitted that such con- tact occurred at the time of preparation or application of the pesticide, and such contact occurred in many parts of the body (64.3% on the hands, 33.3% on the face, 21.1% on the back, 7.1% on the legs and arms, and 2.4% on the neck). These data also indicate the in- correct use of PPE by the participants. The inappropriate use of PPE leaves workers exposed to the absorption of pesticides that can occur through the dermal, respiratory and oral routes, and can lead to acute and/or chronic poisoning (CARGNIN; ECHER; SILVA, 2017). In this sense, the relevance of hygiene measures after the manipulation of pesticides is an im- portant action to reduce the possibility of contamina- tion and absorption of products by the usual routes of entry into the human body. Workers were also asked about training to work with pesticides, as well as handling empty containers (Table 2). Regarding training on the use of pesticides Personal protective equipment N % Simple mask 33 52.4 Mask with filter 25 39.7 Gloves 50 79.0 Boots 57 90.5 Glasses 13 20.6 Coveralls 56 88.9 Impervious apron 25 39.7 Table 1 – Personal protective equipment used by rural workers living in a municipality in western Santa Catarina (2018). http://n.XX Rural workers exposure to organophosphates and carbamates 23 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 and the necessary care in handling them, only 54.0% of workers reported having received information, of which 47.0% stated that the training was carried out by the company that sold the products and 52.9% by other institutions, such as the Municipal Secretariat of Agriculture and the Agricultural Research and Ru- ral Extension Company of Santa Catarina (Empresa de Pesquisa Agropecuária e Extensão Rural de Santa Cata- rina — EPAGRI). The guidelines provided by the selling company basically consisted of the number of products that should be applied to the crops. Adequate technical assessment of the existence of problems related to the attack of insects and diseases harmful to plantations and the real need for the use of pesticides, as well as technical guidance on the use of pesticides as a means of control, are important mea- sures to prevent accidents and pesticide-related poi- sonings (JACOBSON et al., 2009; SIQUEIRA et al., 2013). Handling empty containers also requires special atten- tion. According to NBR 13968 (ABNT, 1997), the con- tainers must be washed three times and returned to the manufacturer. In this sense, 98.4% of the rural workers interviewed stated that they perform the tri- ple washing of empty containers and 68.3% reported discarding the washing water using the product’s own application equipment (Table 2). Although workers were aware of the importance of triple washing and subsequent return of containers to the place where they were purchased, a considerable number of workers (30.2%) declared to discard water in inappropriate places such as close to homes and streams (Table 2). This fact is quite worrying, as these actions imply an increased risk of contamination by other family members, besides domestic animals and water, as observed in previous studies (SIQUEIRA et al., 2013; MIRANDA, 2016). Variables N % Training to work with pesticides Yes 34 54.0 No 29 46.0 Provider of the training* Selling company 16 47.0 Others 18 52.9 Disposal of empty containers Return to the provider 60 95.2 Storage 3 4.8 Triple wash Yes 62 98.4 Place of elimination of triple washing water Spray 43 68.3 Ground/place where it is washed 19 30.2 Others 1 1.3 Table 2 – Characterization of training and management of empty containers by rural workers in a municipality in western Santa Catarina, 2018. *Only workers who said to have received training responded this item. http://n.XX Lorenzatto, L.B. et al. 24 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 The results of the present study also made it possible to know the type of destination given to empty contain- ers. In 95.2% of cases, respondents reported returning the packaging to the place of purchase, according to the law. Moreover, rural workers reported being aware of the existence of the law that determines the correct disposal of pesticide packaging. Adequate disposal is considered to go beyond a con- scious attitude toward compliance with the law, which establishes the rules for the return of empty packaging according to the instructions on the package inserts (Law No. 7,082, of July 11th, 1989, and Decree No. 9,974, of June 6th, 2000). The deadline for disposal is up to one year from the date of purchase. A longer term is only possible if authorized by the regulatory agency (BRASIL, 2000). Failure to comply with this law can result in fines for the farmer, dealer and even the pesticide manufacturer. Types of pesticides used by workers Rural workers were asked about the types of pesticides used. Farmers use an average of 6.4 ± 4.1 types of pes- ticides, with those using less using only one type and those using more using 24 different types. Different formulations were mentioned, including all classes of pesticides, such as insecticides, fungicides, herbicides, poison for ants, termiticides and growth regulators. It is worth mentioning that the participants cited the commercial names of the pesticides and, subsequently, a search was made in the monographs provided by the National Health Surveillance Agency (Agência Nacional de Vigilância Sanitária — ANVISA) on the active ingre- dients (AI), chemical groups, toxicological classification, among other information. Fifty-eight active ingredients were mentioned, and the 10 most cited are described in Table 3. The herbicide glyphosate was the substance most cited by rural workers, and all the interviewees mentioned the use on their farms. Although glyphosate is classi- fied as little toxic (Class IV), it has been in the process of toxicological re-evaluation by ANVISA since 2008 due to indications of its carcinogenic potential and other human health hazards. For this reason, its use will be banned in France from 2022 (BOMBARDI, 2017). It is also worth noting that three of the ten active principles most cited belong to the toxicological classifications I and II (extremely and highly toxic, respectively). Other active ingredients cited by rural workers in the present study, such as 2,4-D and abamectin, paraquat and acephate, which are on the list of the ten best-sell- ing agrochemicals in Brazil, were reassessed by ANVISA, and their use was kept restricted after the evaluation. Active ingredients Chemical group Toxicological classification Glyphosate Substituted glycine Class IV Trifloxystrobin Strobilurin Class II Prothioconazole Triazolinethione Class IV Lambda-cyhalothrin Pyrethroid Class III Methomyl Oxime methyl carbamate Class I Thiamethoxam Neonicotinoid Class III Chlorantraniliprole Anthranilamide Class III 2.4-D Aryloxyalkanoic acid Class I Paraquat Bipyridylium Class III Diurom Urea Class III Table 3 – Active ingredients used by rural workers living in a municipality in western Santa Catarina, 2018. http://n.XX Rural workers exposure to organophosphates and carbamates 25 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 In the European Union (EU), they were banned in 2009 and 2003, respectively. The thiamethoxam insecticide, the sixth most used by the population studied, was also banned in the EU (CARNEIRO, 2015; BOMBARDI, 2017). The active ingredients used by the rural workers partici- pating in the study belonged to the following toxicological classifications: 46.5% class III, 27.6% class II, 13.8% class I; and 12.1% class IV. The use of a high number of active in- gredients belonging to toxicological classes I and II (41.4%) indicates that the population is exposed to highly harmful substances. However, in most cases, these substances can be replaced by less toxic compounds. According to Hess (2018), many products registered with ANVISA belong to toxicological classes III and IV, which meet most of the demands of farmers and with less toxicity. Therefore, the authorized use of highly toxic active substances (classes I and II) is not justified because they can be replaced by products with low- er toxicity. In addition, the main chemical groups used were pyre- throids and triazole, with six ingredients each, followed by organophosphates, carbamates, and strobilurin, with four active ingredients each group, and the neonicoti- noids with three active ingredients. It should also be not- ed that of the 58 active ingredients mentioned, only eight belonged to the chemical groups of organophosphates and carbamates, which cause changes in the activity of the enzyme acetylcholinesterase, with the possibility of acute intoxications. For the other chemical groups used by rural workers, there are no specific biomarkers for the identification of acute or chronic poisoning. Characterization of the effects of pesticides on population health Table 4 shows that the population with occupational exposure to pesticides presented intoxication con- ditions, with 26 signs/symptoms reported after han- dling or applying pesticides. Data analysis showed a significant prevalence of intoxication reported by ru- ral workers (46%). Other Brazilian studies found lower prevalence rates of poisoning, ranging from 11 to 22% (FARIA, 2012; CABRAL, 2012; BURALLI, 2016). Signs and symptoms of poisoning related to exposure to similar pesticides were also found in other studies (CABRAL, 2012; OLIVEIRA, 2017). The signs and symp- toms cited by the workers are linked to acute poisoning (nausea, vomiting, headache, salivation, tremors, men- tal confusion, convulsion, among others) and chronic poisoning (agitation, weakness, respiratory diseases). Some of these symptoms may be related to poisoning by cholinesterase inhibitors. Early symptoms relat- ed to the CNS include headache, dizziness, agitation, tremors, muscle weakness, fatigue, cramps and exces- sive salivation, which is a common muscarinic signal (ALONZO; CORRÊA, 2014). It is important to emphasize that 29 workers (46%) re- ported intoxication, however, when analyzing the symp- toms mentioned (Table 4), 37 workers reported feel- ing discomfort (58.8%), 34 reported headache (54%), and 31 reported nausea (49.2%), these were typical of mild acute poisoning. Buralli (2016) demonstrated in his study that, although a minority of workers reported poi- soning by exposure to pesticides, a considerable number of workers reported having experienced various signs and symptoms characteristic of the poisoning. The fact that farmworkers do not understand these signs as a possible poisoning reveals that the damages are not always perceived, or their occurrence is not given due importance. There is great misinformation among work- ers about the use and dangers of pesticides. They also show to be afraid of recognizing the symptoms of poi- soning, and do not seek medical care or do not admit that the symptoms they have may be related to the use of these substances (LONDRES, 2011). As to the occurrence of health problems, there was a high number (50.8%) of workers with anxiety. It is of fundamental importance to highlight the occurrence of behavioral disorders as an effect of exposure to pesticides. Such disorders include anxiety, irritabili- ty, attention disorders, and sleep disorders. In an ep- idemiological study, Faria et al. (2000) quantitatively demonstrated a strong association between pesticide poisoning and minor psychiatric disorders such as ner- vousness, anxiety, and discouragement. Regarding depression, this study demonstrated that 27.0% of rural workers mentioned this disease and re- ported the use of antidepressants. When compared to the urban male population, the situation is even http://n.XX Lorenzatto, L.B. et al. 26 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 more worrying. A study conducted in an urban pop- ulation estimated that the prevalence of depression was 12.7% among men (PRADO et al., 2012). In this study, the value found for the male population was more than double (27%), suggesting that rural work- ers are more susceptible to the occurrence of depres- sion than urban workers. The high prevalence of signs and symptoms evidenced in the present study indicates that the health of rural Variables N % Reported intoxication 29 46.0 Malaise 37 58.8 Headache 34 54.0 Nausea 31 49.2 Weakness 27 43.0 Salivation 26 41.3 Eye irritation 25 39.7 Shaking 24 38.0 Dizziness 24 38.0 Vomiting 19 30.1 Tremors 19 30.1 Sweating 17 27.0 Colic 15 23.8 Tingling 14 22.2 Blurry vision 14 22.2 Palpitation 13 20.6 Diarrhea 12 19.0 Numbness 11 17.5 Cough 11 17.5 Skin irritation 10 15.9 Shortness of breath 10 15.9 Chest pain 7 11.1 Cramps 5 8.0 Mental confusion 5 8.0 Others 3 4.7 Convulsion 2 3.1 Uncoordinated movements 1 1.6 Table 4 – List of signs and symptoms reported by rural workers residing in a municipality in western Santa Catarina after handling/applying pesticides, 2018. http://n.XX Rural workers exposure to organophosphates and carbamates 27 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 workers is compromised and conditioned to the inten- sive use of pesticides. According to Gomes Filho Neto, Andrade and Felden (2018), understanding the rela- tionship between poisoning and diseases (mental or not) associated with the use of pesticides, is essential for health and occupational safety professionals to act preventively and minimize damage to workers. Expo- sure to pesticides should be seen as a risk determinant for the health of rural workers, and as such, requires public health programs aimed at promoting the health of this population (SILVA, 2013). Evaluation of the erythrocyte acetylcholinesterase enzyme activity According to Ribeiro and Mella (2007), a baseline test is necessary before exposure of workers who will have contact with organophosphate and carbamate pesti- cides, considering that baseline levels of cholinesterase vary from one person to another. The analysis of the activity of the erythrocyte AChE and the comparison of its values before and after exposure are shown in Figure 1. The activity of the AChE enzyme reduced from 0.83 ± 0.06 delta pH/hour in pre-exposure to 0.71 ± 0.11 delta pH/hour in post-exposure (Figure 1). Wilcoxon’s nonparametric test indicated a statistically significant (p < 0.05) decrease in post-exposure AChE values as compared to the pre-expo- sure values. The decrease in enzyme activity (p < 0.05), sug- gests that rural workers were contaminated by pesticides of the organophosphate and carbamate classes. This statistically significant decrease indicates that ru- ral workers exposed to pesticides are vulnerable to the harmful action of these substances. It should be noted that the determination of acetylcholinesterase activi- ty constituted an appropriate indicator to monitor the exposures to these pesticides, but it is not suitable for pesticides from other chemical groups. When the results of the enzymatic dosages of each in- dividual worker were compared, 12 workers (19.6%) were poisoned as indicated by the enzymatic activity with a reduction greater than or equal to 30%, in accor- dance with the NR-7 (BRASIL, 2013). These data indicate that the inadequate management of pesticides by rural workers allowed a significant absorp- tion of organophosphates and carbamates after exposure. Other studies found between 3.6 and 4.6% of the sample presenting a reduction greater than 30% in acetylcholines- terase levels (OLIVEIRA-SILVA et al., 2001; NASCIMENTO, 2013; NASCIMENTO et al., 2013; NERILO et al., 2014). It is important to note that thousands of farmers are intoxicated as a result of exposure to pesticides world- wide (OPAS; OMS, 2018), which has generated great concern for several countries and also for the World Health Organization (KOS et al., 2013). The potential *Statistically significant difference (p < 0.05). Figure 1 – Dosage of erythrocyte acetylcholinesterase enzyme activity in rural workers of a municipality in western Santa Catarina pre- and post-exposure to pesticides, 2018. * Pre-exposure Post-exposure A ch e (D el ta p H /h ou r) 1.0 0.8 0.6 0.4 0.2 0.0 http://n.XX Lorenzatto, L.B. et al. 28 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 REFERENCES ALONZO, H.G.A.; CORRÊA, C.L. Praguicidas. In: OGA, S. et al. (orgs.). Fundamentos de Toxicologia. 4ª ed. São Paulo: Atheneu, 2014. p. 323-341. ANDRADE FILHO, A.; SOUZA, S.D. Anticolinesterásicos. In: ANDRADE FILHO, A.; CAMPOLINA, D.; DIAS, M.B. (orgs.). Toxicologia na Prática Clínica. 2ª ed. Belo Horizonte: Folium, 2013. p. 89-98. ASSOCIAÇÃO BRASILEIRA DE EMPRESAS DE PESQUISA (ABEP). Critério de classificação econômica Brasil. Rio de Janeiro: ABEP, 2015. Disponível em: . Acesso em: 2 jul. 2019. ASSOCIAÇÃO BRASILEIRA DE NORMAS TÉCNICAS (ABNT). NBR 13968: embalagem rígida vazia de agrotóxico: procedimento de lavagem. 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Acesso em: 23 ago. 2019. toxicity of organophosphates and carbamates has been demonstrated in several studies that have shown signif- icant reductions in acetylcholinesterase activity in ex- posed rural workers, making the problem of poisoning by these compounds even more evident (OLIVEIRA-SIL- VA et al., 2001; DHANANJAYAN et al., 2012; MURUSSI et al., 2014; NERILO et al., 2014; KLEIN et al., 2018). In this study, the analysis of erythrocyte acetylcholinester- ase activity indicates that farmworkers are vulnerable to pesticide contamination and may suffer health damage over time. A significant reduction in enzyme activity after exposure indicates inadequate handling of pesticides and unsatisfactory use of protective equipment that allowed an important absorption of these compounds. CONCLUSIONS The results show that exposure to pesticides can have serious health consequences. Acknowledging the com- plexity of the problem and the adoption of more sustain- able and responsible agricultural alternatives are urgently needed in order to reduce the use of these substances and the damage to human health and the environment. CONFLICT OF INTERESTS All authors declare no conflicting interests. ACKNOWLEDGEMENT To the Coordination for the Improvement of Higher Education Personnel (CAPES) and the Community Uni- versity of the Chapecó Region (Unochapecó) for finan- cial support. http://n.XX http://www.abep.org/criterio-brasil http://conexaoagua.mpf.mp.br/arquivos/agrotoxicos/05-larissa-bombardi-atlas-agrotoxico-2017.pdf http://conexaoagua.mpf.mp.br/arquivos/agrotoxicos/05-larissa-bombardi-atlas-agrotoxico-2017.pdf http://renastonline.ensp.fiocruz.br/sites/default/files/arquivos/recursos/Documento%20orientador%20Vig%20Pop%20Agrotox_Vigipeq_completo2013.pdf http://renastonline.ensp.fiocruz.br/sites/default/files/arquivos/recursos/Documento%20orientador%20Vig%20Pop%20Agrotox_Vigipeq_completo2013.pdf https://legis.senado.leg.br/sdleg-getter/documento?dm=3653399&ts=1559239802921&disposition=inline https://legis.senado.leg.br/sdleg-getter/documento?dm=3653399&ts=1559239802921&disposition=inline Rural workers exposure to organophosphates and carbamates 29 RBCIAMB | v.55 | n.1 | mar 2020 | 19-31 - ISSN 2176-9478 BRASIL. 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