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Pertanika J. Trop. Agric. Sc. 42 (1): 61 - 77 (2019) TROPICAL AGRICULTURAL SCIENCE Journal homepage: http://www.pertanika.upm.edu.my/ The Monitoring of Organophosphorus and Carbamate Insecticides and Heavy Metal Contents in Paddy Field Soils, Water and Rice (Oryza sativa L.) Sirikul Thummajitsakul1*, Chanon Sangdee1, Suteema Thaisa1, Penpuck Ruengwiroon1 and Kun Silprasit2 1Division of Health Promotion, Faculty of Physical Therapy, Srinakharinwirot University, Ongkharak, Nakhon- Nayok, 26120, Thailand 2Faculty of Environmental Culture and Ecotourism, Srinakharinwirot University, Bangkok, 10110, Thailand ABSTRACT The aim of this work was to monitor organophosphorus and carbamate insecticides in paddy field soils, water and rice plants by measuring anti-acetylcholinesterase activity, and heavy metal contents by an atomic absorption spectrometry. The results showed that the percentages of anti-acetylcholinesterase activities were found in the order of shoot > soil > root > grain > water. Concentrations of heavy metals in all samples were found in the following order: Fe>Mn>Ni>Pb>Zn, particularly Fe, Ni and Pb were found in concentrations exceeding their maximum permissible levels in all samples of water and rice plants. Interestingly, principal component analysis confirmed positive correlations (significantly:P <0.05) between the percentages of anti-acetylcholinesterase activities and heavy metal contents, between Zn and Mn contents, between Zn and Pb contents, between Pb and Mn contents, and between Pb and Ni contents. Additionally, the translocation factors (TFsoil, TFroot, and TFshoot) and bioaccumulation factor (BAF) above 1 of the rice plant were observed in order of Zn>Fe>Mn>Ni or Pb, Zn or Ni> Pb or Mn >Fe, Pb>Zn or Mn or Fe>Ni, and Zn>Pb>Mn>Ni>Fe, respectively. These data support developing a useful biomarker to monitor organophosphorus and carbamate insecticides and heavy ARTICLE INFO metals, and management programs in Article history: Received: 12 April 2018 health promotion and toxic surveillance for Accepted: 08 October 2018 Published: 26 February 2019 insecticides and heavy metals in food and E-mail addresses: environments. [email protected] (Sirikul Thummajitsakul) [email protected] (Chanon Sangdee) Keywords: Bioaccumulation, heavy metals, [email protected] (Suteema Thaisa) [email protected] (Penpuck Ruengwiroon) organophosphorus and carbamate insecticides, rice [email protected] (Kun Silprasit) * Corresponding author ISSN: 1511-3701 e-ISSN: 2231-8542 © Universiti Putra Malaysia Press Sirikul Thummajitsakul, Chanon Sangdee, Suteema Thaisa, Penpuck Ruengwiroon and Kun Silprasit INTRODUCTION Occupational and Environmental Diseases Nakhon Nayok is one of central provinces [BOED], 2016). Recently, insecticides of Thailand where there are areas for rice of the organophosphorus and carbamate growing of approximately 935.3 square km group are increasingly used to control which provide 311,566 tons of rice products pests, such as insects in agriculture regions. (National Statistical Office [NSO], 2014). However, these compounds can impact the Ongkharak is one of the most important environment and human health. districts of Nakhon Nayok province in The organophosphorus and carbamate rice cultivation. Nowadays, demand of insecticides (e.g., dichlorvos, parathion, agricultural products has continued to fenamiphos, and N-methyl carbamates) increase overtime because of Thailand’s are known as irreversible inhibitors of dramatically growing population. The acetylcholinesterase activity and commonly use of pesticides is therefore essential and used as biomarkers for environmental rapidly increasing. Previously, it has been and human monitoring (Lionetto et al., reported that pesticides were imported to 2013; Vale & Lotti, 2015). Corresponding Thailand in 2016, which was higher than in with previous reports, the estimation of the previous year by approximately 7.54% acetylcholinesterase inhibitory activities (Office of Agricultural Economics [OAE], has been used to monitor or indicate 2017). the presence of organophosphorus and Generally, several chemicals used carbamate insecticides in vegetable and in paddy fields (i.e. inorganic fertilizers fruit juice (Korpraditskul et al., 2004). and pesticides) consist of heavy metals Similarly, the relationship between the (Gimeno-García et al., 1996; Wuana & use of the insecticides and the level of Okieimen, 2011). Therefore, the application acetylcholinesterase in the blood has been of pesticides can lead to extensive significantly found in 236 farmers from distribution and long-term accumulation of Sam Chuk district, Suphan Buri province heavy metals in soils (Jablonowski et al., (Duangchinda et al., 2014). 2012). Heavy metals from soils can enter Therefore, our major goals focused water reservoirs, then accumulate in plants on monitoring organophosphorus and animals, and transfer to humans via food and carbamate insecticides by anti- webs (Gall et al., 2015). Heavy metals are acetylcholinesterase activity, and measuring toxic to human health as they directly effect heavy metal content in soil, water, and rice biochemical and physiological functions from paddy fields in Ongkharak district, in the human body (Jaishankar et al., Nakhon Nayok province by an atomic 2014). Moreover, prevalence rates of toxic absorption spectrometry. Bioaccumulation effect of pesticides have been reported factor (BAF) and translocation factors (TFs) at 14.47 cases per 100,000 people or were also calculated for each rice sample 8,689 cases per year in 2016 (Bureau of and heavy metal. Our hypothesis was that 62 Pertanika J. Trop. Agric. Sc. 42 (1): 61 - 77 (2019) Insecticides and Heavy Metals in Paddy Field Soils, Water and Rice percentages of anti-acetylcholinesterase and chromium) for an atomic absorption activities correlated with each heavy spectrometry were obtained from Sigma- metal content in rice plants. These basic Aldrich. data are useful for promoting awareness and surveillance of human health and Sampling environmental impacts. These benefits help Samples from paddy soils (N= 30), rice to reduce the insecticide use of farmers in (N=30) and water (N=10) were collected paddy fields, and to manage programs in from 10 paddy fields located in Tambon sustainable prevention and treatment of Ongkharak (OK), Khlong Yai (KY), Sisa human diseases from insecticides and heavy Krabue (SK), Bang Luk Suea (BLS), Sai metals in food. Mun (SM), Bang Plakot (BP), Chumphon (CP), Bueng San (BS), Phra Achan (PA), MATERIALS AND METHOD and Bang Sombun (BSB) of Ongkharak Chemicals district, Nakhon Nayok province between Acetylthiocholine iodide was purchased August 2017 and February 2018. Sampling from Sigma. Acetylcholinesterase enzyme, locations were demonstrated with GPS DTNB (5,5′-dithiobis, 2-nitrobenzoic acid) (Global Positioning System) using Google and nitric acid were obtained from Sigma- Earth (version 7.1.2.2041) (Figure 1). Aldrich. Absolute ethanol was obtained Water samples were collected by the from Merk. Heavy metal standards (copper, grab sampling method at approximately 15 lead, ferrous, nickel, zinc, manganese, cm depth from the water surface, then kept in Figure 1. Sampling locations were revealed with GPS (Global Positioning System) using Google Earth (version 7.1.2.2041). Tambon Ongkharak = OK, Khlong Yai = KY, Sisa Krabue = SK, Bang Luk Suea = BLS, Sai Mun =SM, Bang Plakot = BP, Chumphon =CP, Bueng San =BS, Phra Achan =PA, and Bang Sombun =BSB. Pertanika J. Trop. Agric. Sc. 42 (1): 61 - 77 (2019) 63 Sirikul Thummajitsakul, Chanon Sangdee, Suteema Thaisa, Penpuck Ruengwiroon and Kun Silprasit 50 ml polyethylene tubes at 4oC (Chowdhur at 410 nm by spectrophotometer (Model et al., 2012; Thummajitsakul et al., 2015). T60UV). Negative controls for quality Soil subsamples from 10 sites were collected control procedures were carried out using at 15 depth from the surface by sampling distilled water, and each sample extract tools, then combined into one sample, and without acetylcholinesterase enzymes. each sample (approximately 500 grams) was Methylcarbamate was used as positive used for drying at 50oC before preserving in control at concentrations 0, 0.2, 2, 20 and 200 plastic bags at a temperature of 4oC (Fery µg/ml. Percentages of acetylcholinesterase & Murphy, 2013; Thummajitsakul et al., inhibition were calculated following the 2015). For rice sampling, each subsample below formula. was collected from 10 sites per a paddy field, % Acetylcholinesterase inhibition = then combined into one sample, and divided into three parts (root, shoot and grain) [(Awater – Ablank1) – (Asample – Ablank2)]*100 according to the report of Kingsawat and (A – A ) Roachanakanan (2011). The different parts water blank1 of the rice plant were cleaned with deionized Note: water, then sectioned into small pieces, and (a) Awater and Asample were the absorbance dried at 50oC, followed by grinding with a of distilled water and sample extract homogenizer and preserving the powdered with acetylcholinesterase enzyme, samples in plastic bag at 4o C. respectively. (b) Ablank1 and Ablank2 were the absorbance Anti-acetylcholinesterase Activity of distilled water and sample The anti-acetylcholinesterase activity extract without acetylcholinesterase was performed using Ellman colorimetric enzyme, respectively. assay (Ellman et al., 1961) with some modifications. Each powdered sample Digestion and Sample Analysis (0.5 grams for rice samples and 2.5 grams Sample digestion was performed according for soil samples)