Tobacco is an important cash crop of Pakistan. Pesticides are commonly used to increase the crop yield, but their health impact has not been studied yet. The objectives of the study were to determine the frequency of pesticide poisoning and to explore the knowledge, attitudes and practices (KAP) towards safety measures among the tobacco farmers in Swabi, Pakistan. One hundred and five tobacco farmers involved in pesticide application were randomly selected from two villages of district Swabi. A structured questionnaire was used for clinical and KAP information. Plasma cholinesterase (PChE) levels were measured by Ellman's method by using GD Italy kits. All tobacco farmers were males with a mean (SD) age of 26 (9) years. The majority of the farmers reported multiple symptoms headache, dizziness, vomiting, shortness of breath, muscle weakness and skin rash correlate with the clinically significant depression of PChE levels. Out of 105 pesticide applicators, 58 (55%) had post-exposure reduction in PChE levels o20% from baseline, 35 (33%) had mild poisoning (20-40% reduction) and 12 (11%) had moderate poisoning (440% reduction). Most of the farmers did not use any personal protective equipment during pesticide handling. Only a few used shoes (31%), masks (14%) and gloves (9%) during pesticide spray. In conclusion, the tobacco farmers had mild to moderate pesticide poisoning, which was correlated with depression in PChE levels. Moreover, most farmers had little knowledge about the safety measures, casual attitude and unsatisfactory safety practices with regard to the use of basic protective equipments during pesticide applications on the tobacco crop.

The study aimed to determine the hazardous health effects of pesticides exposure in the factory workers by measuring plasma cholinesterase (PChE), pesticides residues, and renal and hepatic biochemical markers. In addition, we also assessed the knowledge, attitudes, and safety practices adopted by the industrial workers. The study was conducted in three different sizes of factories located in Lahore (large), Multan (medium), and Karachi (small) in Pakistan. Total 238 adult males consisting of 184 pesticide industrial workers (exposed group) from large-sized (67), medium-sized (61), small-sized (56) industrial formulation factories, and 54 controls (unexposed) were included in the study. All the participants were male of aged 18 to 58 years. PChE levels were estimated by Ellmann's method. Plasma pesticides residue analysis was performed by using reverse phase C-18 on high-performance liquid chromatograph and GC with NPD detector. Plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine, urea, and gamma glutamyltransferase (GGT) were measured on Selectra E auto analyzer. Plasma and C-reactive protein was analyzed by Immulite 1000. The results revealed a significant decrease in plasma post exposure PChE levels (<30%) as compared to baseline in the workers of small (29%) and medium (8%) industrial units (p < 0.001). Plasma cypermethrin, endosulfan, imidacloprid, thiodicarb, carbofuran, and methamidophos levels were found to be higher than allowable daily intake. Serum AST, ALT, creatinine GGT, malondialdehyde, total antioxidant, and CRP were significantly raised among the workers of small and medium pesticide formulation factories as compared to large industrial unit and controls (p < 0.001). The study demonstrated that unsafe practices among small- and medium-sized pesticides industrial workers cause significant increase in pesticide exposure, oxidative stress, and derangement of hepatic and renal function.

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Phosphotriesterases (PTEs) have been isolated from a range of bacterial species, including Agrobcaterium radiobacter (PTEAr), and are efficient enzymes with broad substrate ranges. The turnover rate of PTEAr for the common organophosphorous insecticide malathion is lower than expected based on its physical properties; principally the pka of its leaving group. In this study, we rationalise the turnover rate of PTEAr for malathion using computational docking of the substrate into a high resolution crystal structure of the enzyme, suggesting that malathion is too large for the PTEAr binding pocket. Protein engineering through combinatorial active site saturation testing (CASTing) was then used to increase the rate of malathion turnover. Variants from a CASTing library in which Ser308 and Tyr309 were mutated yielded variants with increased activity towards malathion. The most active PTEAr variant carried Ser308Leu and Tyr309Ala substitutions, which resulted in a ca. 5000-fold increase in k cat/K M for malathion. X-ray crystal structures for the PTEAr Ser308Leu\Tyr309Ala variant demonstrate that the access to the binding pocket was enhanced by the replacement of the bulky Tyr309 residue with the smaller alanine residue.

Malathion, a well-known organophosphate pesticide, has been used in agriculture over the last two decades for controlling pests of economically important crops. In the present study, a single bacterium, ML-1, was isolated by soilenrichment technique and identified as Bacillus licheniformis on the basis of the 16S rRNA technique. The bacterium was grown in carbon-free minimal salt medium (MSM) and was found to be very efficient in utilizing malathion as the sole source of carbon. Biodegradation experiments were performed in MSM without carbon source to determine the malathion degradation by the selected strain, and the residues of malathion were determined quantitatively using HPLC techniques. Bacillus licheniformis showed very promising results and efficiently consumed malathion as the sole carbon source via malathion carboxylesterase (MCE), and about 78% malathion was degraded within 5 days. The carboxylesterase activity was determined by using crude extract while using malathion as substrate, and the residues were determined by HPLC. It has been found that the MCE hydrolyzed 87% malathion within 96 h of incubation. Characterization of crude MCE revealed that the enzyme is robust in nature in terms of organic solvents, as it was found to be stable in various concentrations of ethanol and acetonitrile. Similarly, and it can work in a wide pH and temperature range. The results of this study highlighted the potential of Bacillus licheniformis strain ML-1 as a biodegrader that can be used for the bioremediation of malathion-contaminated soil.

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This study is based on self-reported information collected from selected farmers of Vehari District, Punjab, Pakistan, to determine their technical knowledge and awareness about pesticide use and associated environmental and health risks. Moreover, soil contamination by routinely used persistent organochlorine pesticide, endosulfan, was also evaluated. Survey data revealed very low literacy rate (on an average 9th grade education) and technical knowledge (almost missing) of the farmers in Vehari District. The farmers are unable to fully read and understand the instructions about the use of pesticide marked on the containers. They are not fully aware of pesticide persistence and toxicity (73%), unable to identify cotton pests and diseases (86%), and do not know which crop to grow in cotton adjacent fields (100%). Data also revealed that the farmers (100%) do not follow safety measure during pesticide application and are unaware of pesticide toxicity symptoms in human as well as the basic first-aid practices (89%).Poor literacy rate and lack of technical knowledge of farmers in Vehari regarding pesticide use and handling are posing serious environmental and health risks among the local inhabitants, particularly among farmers. Soil analysis results showed that concentration of α- and β-endosulfan ranged from 0-14 to 0-14.64 μg/mg, respectively. Principal component analysis showed that soil organic matter is the key soil parameter controlling the occurrence and fate of endosulfan under sandy loam soil conditions of Vehari District. There is a serious need of improving technical and environmental knowledge of farmer about pesticide risks on human health in the studied area, in particular, and the entire country in general. Findings are of great use for policymaking in Pakistan to minimize pesticide risks in Pakistan.