Human health risk assessment of organophosphorus pesticides in maize (Zea mays L.) from Yushu,Northeast China

In order to investigate organophosphorus pesticides (OPs) concentrations in maize and estimate the health risk to consumers, a total of 55 samples were collected from Yushu, one of the most important maize production areas. The concentrations of the eleven detected OPs in maize were analyzed by gas chromatography coupled with flame photometric detector (GC-FPD). The results showed that OPs concentrations of all maize were not higher than maximum residue limit (MRL), 67.3% of samples below MRL and only in 32.7% of samples was not found OPs. The mean concentrations obtained for the eleven OPs in μg kg^-1 were as follows: omethoate (0.8), quinalphos (0.8), phorate (0.7), dimethoate (0.7), parathion-methyl (0.6), isocarbophos (0.6), diazinon (0.5), fenitrothion (0.5), and malathion (0.5), with parathion (0.5) and fenthion (0.3). Phorate (16.4%), dimethoate (16.4%) and quinalphos (16.4%) had the highest frequency in the eleven OPs. 29.1% samples contained two or more kinds of OPs, while 38.2% samples detected one kind of OPs. The target hazard quotients (THQ) values were all less than one and the total acute hazard index (aHI) values for adults and children were 0.051 and 0.121, respectively indicating that consumer may not pose significant chronic and acute health risk through maize.     

Survey of fungal counts and natural occurrence of aflatoxins in Malaysian starch-based foods

In a survey of starch-based foods sampled from retail outlets in Malaysia, fungal colonies were mostly detected in wheat flour (100%), followed by rice flour (74%), glutinous rice grains (72%), ordinary rice grains (60%), glutinous rice flour (48%) and corn flour (26%). All positive samples of ordinary rice and glutinous rice grains had total fungal counts below 103 cfu/g sample, while among the positive rice flour, glutinous rice flour and corn flour samples, the highest total fungal count was more than 103 but less than 104 cfu/g sample respectively. However, in wheat flour samples total fungal count ranged from 102 cfu/g sample to slightly more than 104 cfu/g sample. Aflatoxigenic colonies were mostly detected in wheat flour (20%), followed by ordinary rice grains (4%), glutinous rice grains (4%) and glutinous rice flour (2%). No aflatoxigenic colonies were isolated from rice flour and corn flour samples. Screening of aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2 using reversed-phase HPLC were carried out on 84 samples of ordinary rice grains and 83 samples of wheat flour. Two point four percent (2.4%) of ordinary rice grains were positive for aflatoxin G1 and 3.6% were positive for aflatoxin G2. All the positive samples were collected from private homes at concentrations ranging from 3.69–77.50 μg/kg. One point two percent (1.2%) of wheat flour samples were positive for aflatoxin B1 at a concentration of 25.62 μ};g/kg, 4.8% were positive for aflatoxin B2 at concentrations ranging from 11.25–252.50 μg/kg, 3.6% were positive for aflatoxin G1 at concentrations ranging from 25.00–289.38 μg/kg and 13.25% were positive for aflatoxin G2 at concentrations ranging from 16.25–436.25 μg/kg. Similarly, positive wheat flour samples were mostly collected from private homes. This revised version was published online in June 2006 with corrections to the Cover Date.     

Determination of organophosphorus pesticide residues in human tissues by capillary gas chromatography-negative chemical ionization mass spectrometry analysis

We describe an analytical method that allows the determination of organophosphorus pesticides (OPs) in different human tissues. It involves an extraction procedure with ethanol-ethyl acetate, followed by gel permeation chromatography clean-up step and analysis by capillary gas chromatography-negative chemical ionization mass spectrometry in the selected ion monitoring mode. The method was tested for 37 OPs and the recoveries obtained vary between 60 and 106% with standard deviations ranging between +/-2 and +/-10. These values are independent of the analyzed tissue. Peak area repeatability as RSD for some OPs was < or =4.8% while a good linear relationship in the range 1.0-500 pg microl(-1) with r(2)> or =0.9878 was obtained. The limit of detection for the 37 OPs falls between 0.01 and 0.09 ng g(-1) with an RSD< or =9.5%. The analytical set up in this paper has been used to analyze different samples of human tissues (liver, healthy kidney, cancer kidney and adipose tissue) of 24 patients. The number of the identified OPs in the tissue samples is different (max. 20) according to the sample while their concentration ranges between the limit of detection and 28.0 ng g(-1). The highest concentrations have been determined in liver samples without any pathology (0.4-28.0 ng g(-1)) while the lowest concentrations have been determined in healthy kidney samples (0.01-1.50 ng g(-1)). In the cancer kidney samples OP concentrations vary between 0.03 and 4.6 ng g(-1): these concentrations are more elevated than those determined in healthy kidney samples. The comparison between the concentration of OPs determined in the healthy part, when possible, and those determined in the cancer part of the same kidney sample are very interesting: in fact, in the latter the OP concentration is generally 1-2-times higher than that in the former, an index of lower enzymatic activity in the cancer tissue.     

Mapping quantitative trait loci associated with arsenic accumulation in rice (Oryza sativa)

The quantitative trait loci (QTLs) associated with arsenic (As) accumulation in rice were mapped using a doubled haploid population established by anther culture of F1 plants from a cross between a Japonica cultivar CJ06 and an Indica cultivar TN1 (Oryza sativa). Four QTLs for arsenic (As) concentrations were detected in the map. At the seedling stage, one QTL was mapped on chromosome 2 for As concentrations in shoots with 24.4% phenotypic variance and one QTL for As concentrations in roots was detected on chromosome 3. At maturity, two QTLs for As concentrations in grains were found on chromosomes 6 and 8, with 26.3 and 35.2% phenotypic variance, respectively. No common loci were detected among these three traits. Interestingly, the QTL on chromosome 8 was found to be colocated for As concentrations in grain at maturity and shoot phosphorus (P) concentrations at seedling stage. These results provide an insight into the genetic basis of As uptake and accumulation in rice, and will be useful in identifying genes associated with As accumulation.     

Assessment of heavy metal contamination of rice grains (Oryza sativa) and soil from Ada field,Enugu, Nigeria: Estimating the human healtrisk

The concentrations of heavy metals in rice grains and soils from Ada cultivated fields were investigated. Rice and soil samples were digested and the heavy metal concentrations determined using atomic absorption spectrophotometer. The results showed the following concentrations of metals (mg/kg): soil—Pb (4.64 ± 2.18), Cd (0.83 ± 0.83), Zn (20.26 ± 18.60), Mn (68.90 ± 19.91), Ni (3.46 ± 2.42), and Cr (21.41 ± 14.6); rice—Pb (3.99 ± 1.43), Cd (1.10 ± 0.53), Zn (65.37 ± 58.09), Mn (37.81 ± 5.82), Ni (3.12 ± 1.49), and Cr (10.87 ± 6.47). The Canadian, Nigerian and Chinese maximum allowable concentration for cadmium in soil were exceeded by 15%, 30%, and 85% of the soil samples, respectively. Heavy metals in all the rice samples evaluated were found to be above the World Health Organization (WHO) maximum permissible limit for lead, cadmium, and chromium. Strong positive and significant correlations were observed between some metal pairs in soil and rice indicating the similarity in origin. The estimated daily intakes of Pb and Cd from rice grown on the fields were higher than the safety levels established by WHO and the Joint FAO/WHO Expert Committee Food Additive, respectively. Hazard quotients and total hazard index for Pb and Cd were greater than 1. This indicates that consumption of rice from these fields will likely induce adverse health effects arising largely from Pb and Cd exposure.     

Genetic information from progeny trials: a comparison between progenies generated by open pollination and by controlled crosses

When conifer progenies generated by open pollination are assessed in field tests, it is usually assumed that all progenies of the same mother are true half-sibs. This assumption may be invalid, leading to overestimation of additive genetic variation and heritability and to biased breeding values. From one Scots pine (Pinus sylvestris) and one Norway spruce (Picea abies) seed orchard, containing 28 and 36 parent clones, respectively, progenies generated by open pollination (OP) and by controlled crosses (CC) were planted in adjacent trials at two to three sites in southern Sweden. The tree height and diameter at breast height were measured, and genetic parameters based on these traits were estimated for OP and CC progenies separately, in order to enable comparisons. Narrow-sense heritability estimates for Scots pine and Norway spruce OP progenies (in the ranges 0.04–0.13 and 0.15–0.38, respectively) did not differ significantly from CC estimates (0.07–0.12 and 0.23–0.30), suggesting that OP-based heritability values were not overestimated to any great extent. Similarly, genetic correlations between OP and CC progenies were in the ranges of 0.87–0.88 and 0.74–0.77 for Scots pine and Norway spruce, respectively, being significantly lower than unity only in the case of Norway spruce. OP-based breeding values for both species should therefore correspond well with those predicted from CC progenies, albeit not perfectly for Norway spruce. In conclusion, the assumption of true half-sibs for OP progenies was not violated to the extent that genetic parameter estimates or breeding value predictions were seriously biased.     

韭菜上五种有机磷农药的残留监测与安全性评价

【目的】2013—2016年,在北京、天津、河北和山东采集韭菜样品,采用超高效液相色谱-三重四级杆串联质谱法检测甲胺磷、甲拌磷、甲基对硫磷、对硫磷和毒死蜱的残留量,并对其残留安全性进行评价。【方法】韭菜样品用乙腈提取,石墨碳固相萃取小柱净化,超高效反相液相色谱分离,三重四级杆质谱检测,基质匹配标准溶液的外标法定量,在添加浓度为0.01~5.0 mg/kg时,方法的准确度、灵敏度和精密度满足农药残留检测的要求。【结果】甲胺磷的最高检出浓度由70.51 mg/kg降低到0.03 mg/kg,超标率由38.10%降到2.03%;甲拌磷的最高检出浓度由6.45 mg/kg降低到0.04 mg/kg,超标率由23.81%降到1.35%;甲基对硫磷的最高检出浓度由6.62 mg/kg降低到0.13 mg/kg,超标率由25.40%降到2.70%;毒死蜱的最高检出浓度由3.95 mg/kg降低到0.47 mg/kg,超标率由15.87%降到4.73%;对硫磷的最高检出浓度由22.69mg/kg降低到0.03 mg/kg,超标率由15.08%降到1.35%。【结论】上述农药的最高残留检出浓度和超标率大幅降低,但是韭菜中仍能检出少量残留。应加强安全综合防治技术的研发,加强安全用药的宣传和指导,保障韭菜产品安全。     

A theoretical model of the competition between hydrolase and carboxylesterase in protection against organophosphorus poisoning

A system of coupled non-linear differential equations describing interactions between organophosphorus compounds (OPs), OP hydrolase, acetylcholinesterase (AChE), and carboxylesterase (CaE) in a single compartment was derived incorporating irreversible combination of OP with AChE, hydrolytic breakdown of OP, and irreversible combination of OP with CaE. The equations were then uncoupled, providing non-linear differential equations on AChE, CaE and OP concentrations. One steady state solution of the AChE equation provided theoretical expressions for the amounts of OP hydrolyzed, bound with CaE, and bound with AChE. Assuming that the LD50 of an OP reflects the dose that depletes AChE to a 'minimal essential' level and that a single compartment model is applicable in vivo, the steady state solution becomes an equation predicting the LD50 from rate constants, initial enzyme levels, and the allowable AChE depletion. Normalization by initial AChE concentration produced a dimensionless relationship describing an 'OP toxicity surface' that clearly demonstrates regions where hydrolysis and CaE offer protection against OP poisoning. The surface can be used to theoretically predict an LD50 given only kinetic rate constants and effective whole-body AChE and CaE levels. Predictions of LD50s of seven OPs in rats were compared with published data. The relationship was found to adequately predict published LD50s spanning 5 orders of magnitude. The OP toxicity surface relationship provides a conceptual tool for use in OP toxicity research but should be particularly useful in predicting the relative protective effects of catalytic and stoichiometric scavenger mechanisms for an OP.     

Isolation of bacterial strains that produce the endocrine disruptor,octylphenol diethoxylates,in paddy fields

Topsoil samples were collected from 36 different paddy fields in West Japan. Each soil sample was incubated with a basal salt-medium containing 0.2% OPPEO. Twelve samples possessed OPPEO-degrading activity, from which twelve cultures of OPPEO-degrading bacteria were isolated. The isolated bacteria grew on a medium containing 0.2% OPPEO as the sole carbon source, and OP2EO and OP3EO were accumulated in the medium under aerobic conditions. OP1EO and octylphenol, which have often been identified in surface water together with OP2EO, were not observed in this experiment. The bacterial isolates were gram negative and tentatively identified as Pseudomonas putida (10 isolates) and Burkholderia cepacia (one isolate) by BIOLOG and 16S rDNA RFLP analyses.     

Heavy metal contamination and health risk assessment in soil-rice system near Xinqiao mine in Tongling city,Anhui province,China

In this study, paddy soil and rice grain samples were collected from the vicinity around the Xinqiao mine in Tongling, China to test for the presence of heavy metals (Cd, Ni, Cr, Cu, Zn, and Pb) in soil-rice system. Results indicated that the soil samples were primarily contaminated with Cd and Cu and followed with Zn and Pb. In rice grains, Cd, Cu, and Cr concentrations exceeded recommended guidelines. However, the regional distribution of heavy metals in rice grains and soil was inconsistent. The bioaccumulation factor of heavy metals in rice grains decreased in the order of Cd > Zn > Cu > Ni > Cr > Pb. The BAF was significantly positively correlated with TCLP-extractable metals and significantly negatively correlated with soil pH. However, the relationship between soil organic matter and the BAF in rice grains was complex. Health risk assessment through rice intake showed that hazard quotients of Cu and Cd were greater than 1 and could pose a considerable non-cancer health risk to adults and children; meanwhile, Cr, Ni, and Cd could pose an unacceptable cancer risk. The results indicated that the government must take measures to reduce heavy metal contents in paddy soil and rice.     

Polymorphism in the acetylcholinesterase gene of Musca domestica L. field populations in Turkey

Acetylcholinesterase (AChE), encoded by the Ace gene, is the primary target of organophosphates (OPs) and carbamates (CBs) in insects. Ace mutations have been identified in OP and CB resistant strains of Musca domestica. In this study, the Ace gene was partially amplified and sequenced at amino acid positions 260, 342, and 407 to determine the frequencies of these mutations in housefly samples collected from farms and garbage disposal sites of 16 provinces in the Aegean and Mediterranean regions of Turkey. In addition, the percent remaining AChE activities in these samples were assayed by using three OPs (malaoxon, paraoxon, and dichlorvos) and one CB (carbaryl) compound as inhibitors. In all the analyzed samples, 13 different combinations at the three amino acid positions were identified and the L/V260-A/G342-F/Y407 combination was found in the highest frequency. No susceptible individual was detected. The highest mean percent remaining AChE activities were detected in the individuals having the L260-A/G342-F/Y407 genotype when malaoxon and paraoxon were used as inhibitors and in the individuals with the L260-A342-F/Y407 combination when dichlorvos and carbaryl were used as inhibitors. The obtained data were heterogeneous and there was no exact correlation between the molecular genetic background and the resistance phenotypes of the flies. The findings of this study at the molecular and biochemical levels indicate the presence of significant control problems in the field.     

Accumulation of ACE Inhibitory Tripeptides,Val-Pro-Pro and Ile-Pro-Pro,in Vascular Endothelial Cells

Topsoil samples were collected from 36 different paddy fields in West Japan. Each soil sample was incubated with a basal salt-medium containing 0.2% OPPEO. Twelve samples possessed OPPEO-degrading activity, from which twelve cultures of OPPEO-degrading bacteria were isolated. The isolated bacteria grew on a medium containing 0.2% OPPEO as the sole carbon source, and OP2EO and OP3EO were accumulated in the medium under aerobic conditions. OP1EO and octylphenol, which have often been identified in surface water together with OP2EO, were not observed in this experiment. The bacterial isolates were gram negative and tentatively identified as Pseudomonas putida (10 isolates) and Burkholderia cepacia (one isolate) by BIOLOG and 16S rDNA RFLP analyses.     

A simple, label-free AuNPs-based colorimetric ultrasensitive detection of nerve agents and highly toxic organophosphate pesticide

Here, a simple label-free colorimetric sensing method for organophosphate (OP) nerve agents and pesticide based on catalytic reaction of acetylcholine esterase (AChE) and the aggregation of lipoic acid (LA) capped AuNPs has been established, which is highly sensitive with a limit of detection (LOD) lowered to pM level. In this method, only the AChE hydrolysis product of acetylthiocholine (ATCh), i.e., cationic thiocholine (TCh) can induce the aggregation of LA capped AuNPs along with a distinct color change from red to steel-blue. When OPs as enzyme inhibitors exist, the generation of TCh can be suppressed and the color change of LA capped AuNPs is gradually diminished according to different concentrations of OPs. The feasibility of this method has been demonstrated by sensitive measurement of OP nerve agents and pesticide in a spiked fruit sample with reliable results. This distinct and rapid colorimetric response enables us to readily probe OPs without more technical demand.     

A preliminary health risk assessment of heavy metals in local and imported rice grains marketed in Trinidad and Tobago,W.I.

Abstract

Within recent times, heavy metal contamination in rice (Oryza Sativa) has been of growing concern particularly due to its implications on human health via consumption. Twenty-one different brands of rice marketed in Trinidad and Tobago were investigated for their heavy metal content (Cd, Cr, Cu, Mn, Ni, Pb and Zn) using Flame Atomic Absorption Spectroscopy. Lead was not detected in any of the samples, while both Cadmium and Nickel were detected in only one sample each at 0.27 and 1.79?µg/g, respectively. The average metal concentrations decreased in the following order: Zinc?> Manganese?> Copper?>?Chromium with concentrations ranging from 1.00–4.35, 1.72–29.92, 5.87–43.56 and 0.06–1.16?µg/g, respectively. The target hazard quotient (THQ) and hazard index (HI) values for all samples were less than one, indicating no risk of non-carcinogenic effects to consumers.     

Cytogenetic response without changes in peripheral cholinesterase enzymes following exposure to a sheep dip containing diazinon in vivo and in vitro

Occupational exposure to organophosphorus insecticides (OPs), such as diazinon, may be monitored by the measurement of the activity of peripheral cholinesterase enzymes, including erythrocyte acetylcholinesterase (EAChE) and plasma or serum cholinesterase (plasma or serum ChE). Exposures have also been measured by the analysis of dialkyl phosphate metabolites of OPs in urine. The potential health risks associated with exposure, especially those of a neurological nature, may then be estimated, and appropriate measures to reduce or eliminate exposures can be implemented. There is evidence that some OP pesticides may have in vivo genotoxic effects, suggesting a possible link with cancer with long term or repeated heavy exposures. This paper describes work performed in 17 subjects with a single or two exposures to a sheep dip containing diazinon. Urine samples revealed OP metabolites dimethylphosphate (DMP), dimethylthiophosphate (DMTP), diethylphosphate (DEP) and diethylthiophosphate (DETP) in 37% of subjects at low levels which were not elevated after exposure. EAChE and plasma ChE were also unchanged before and after exposure, and were similar to those measured in unexposed control groups. Sister chromatid exchanges (SCE), a marker of chromosome damage, was significantly elevated in peripheral blood lymphocytes after exposure compared with before. SCE were unchanged in a group of non-occupationally exposed workers. In vitro studies with both authentic diazinon (98%) and diazinon in a sheep dip formulation (45%) showed increased SCE and decreased replicative indices, suggesting toxic and genotoxic effects of diazinon.     

Direct effects of organic pollutants on the growth and gene expression of the Baltic Sea model bacterium Rheinheimera sp. BAL341

Organic pollutants (OPs) are critically toxic, bioaccumulative and globally widespread. Moreover, several OPs negatively influence aquatic wildlife. Although bacteria are major drivers of the ocean carbon cycle and the turnover of vital elements, there is limited knowledge of OP effects on heterotrophic bacterioplankton. We therefore investigated growth and gene expression responses of the Baltic Sea model bacterium Rheinheimera sp. BAL341 to environmentally relevant concentrations of distinct classes of OPs in 2-h incubation experiments. During exponential growth, exposure to a mix of polycyclic aromatic hydrocarbons, alkanes and organophosphate esters (denoted MIX) resulted in a significant decrease (between 9% and 18%) in bacterial abundance and production compared with controls. In contrast, combined exposure to perfluorooctanesulfonic acids and perfluorooctanoic acids (denoted PFAS) had no significant effect on growth. Nevertheless, MIX and PFAS exposures both induced significant shifts in gene expression profiles compared with controls in exponential growth. This involved several functional metabolism categories (e.g. stress response and fatty acids metabolism), some of which were pollutant-specific (e.g. phosphate acquisition and alkane-1 monooxygenase genes). In stationary phase, only two genes in the MIX treatment were significantly differentially expressed. The substantial direct influence of OPs on metabolism during bacterial growth suggests that widespread OPs could severely alter biogeochemical processes governed by bacterioplankton.     

Cytogenetic response without changes in peripheral cholinesterase enzymes following exposure to a sheep dip containing diazinon in vivo and in vitro

Occupational exposure to organophosphorus insecticides (OPs), such as diazinon, may be monitored by the measurement of the activity of peripheral cholinesterase enzymes, including erythrocyte acetylcholinesterase (EAChE) and plasma or serum cholinesterase (plasma or serum ChE). Exposures have also been measured by the analysis of dialkyl phosphate metabolites of OPs in urine. The potential health risks associated with exposure, especially those of a neurological nature, may then be estimated, and appropriate measures to reduce or eliminate exposures can be implemented. There is evidence that some OP pesticides may have in vivo genotoxic effects, suggesting a possible link with cancer with long term or repeated heavy exposures. This paper describes work performed in 17 subjects with a single or two exposures to a sheep dip containing diazinon. Urine samples revealed OP metabolites dimethylphosphate (DMP), dimethylthiophosphate (DMTP), diethylphosphate (DEP) and diethylthiophosphate (DETP) in 37% of subjects at low levels which were not elevated after exposure. EAChE and plasma ChE were also unchanged before and after exposure, and were similar to those measured in unexposed control groups. Sister chromatid exchanges (SCE), a marker of chromosome damage, was significantly elevated in peripheral blood lymphocytes after exposure compared with before. SCE were unchanged in a group of non-occupationally exposed workers. In vitro studies with both authentic diazinon (98%) and diazinon in a sheep dip formulation (45%) showed increased SCE and decreased replicative indices, suggesting toxic and genotoxic effects of diazinon.     

Cadmium and lead in rice grains and wheat breads in Isfahan (Iran) and human health risk assessment

This study was conducted to survey the levels of cadmium (Cd) and lead (Pb) in the main foods of Isfahan province (central Iran). The rice grains and wheat breads were collected from Isfahan city and villages around the Isfahan and Mobarakeh Steel companies. The food samples were wet-ashed by heating in the presence of concentrated nitric acid and hydrogen peroxide. The concentrations of Cd and Pb were determined by inductively coupled plasma-optical emission spectrometer (ICP-OES). The potential exposure risks were then estimated. Levels of Cd in the rice grains and Pb in the investigated foods (except Sangak beard) were above the maximum levels as set by the Codex Alimentarius and National standards of Iran. The wheat bread samples in Isfahan city and Mobarakeh Steel regions presented higher estimated weekly intake (EWI) of Pb (40.93 and 35.35 µg kg^–1 body weight, respectively) compared to other foods. The EWI for Cd and Pb in most samples was considerably above than the provisional tolerable weekly intake. In most cases, the target hazard quotients values were above one and consequently the consumption of foods in the investigated regions was a potentially serious health risk caused by exposure to the crops contaminated with Cd and Pb.     

Paraoxonase (PON 1) as a biomarker of susceptibility for organophosphate toxicity

Paraoxonase (PON1) is an A-esterase capable of hydrolysing the active metabolites (oxons) of a number of organophosphorus (OP) insecticides such as parathion, diazinon and chlorpyrifos. PON1 activity is highest in liver and plasma, and among animal species significant differences exist, with birds and rabbits displaying very low and high activity, respectively. Human PON1 has two polymorphisms in the coding region (Q192R and L55M) and five polymorphisms in the promoter region. The Q192R polymorphism imparts different catalytic activity toward some OP substrates, while the polymorphism at position -108 (C/T) is the major contributor to differences in the level of PON1 expression. Animal studies have shown that PON1 is an important determinant of OP toxicity, with animal species with a low PON1 activity having an increased sensitivity to OPs. Administration of exogenous PON1 to rats or mice protects them from the toxicity of OPs. PON1 knockout mice display a high sensitivity to the toxicity of diazoxon and chlorpyrifos oxon, but not paraoxon. In vitro assayed catalytic efficiencies of purified PON₁₉₂ isoforms for hydrolysis of specific oxon substrates accurately predict the degree of in vivo protection afforded by each isoform. Low PON1 activity may also contribute to the higher sensitivity of newborns to OP toxicity.