Plasma cholinesterase activity as an environmental impact biomarker in juvenile green turtles (Chelonia mydas) / Atividade da colinesterase plasmática como biomarcador de impacto ambiental em tartarugas verdes juvenis (Chelonia mydas)

The objective of this study was to evaluate the enzymatic activity of plasma cholinesterase in Chelonia mydas marine turtles belonging to two populations, according to their capture sites, under the absence and probable influence of anthropic effects. A total of 74 animals were used and later divided into two groups, based on the capture site. Blood samples were collected from all captured animals, which were then released into the sea at the site of capture. A descriptive statistical analysis of the plasma cholinesterase activity values and an analysis comparing these values based on the capture site were performed. Samples of heparinized plasma from animals captured at the two different sites were analyzed. Plasma cholinesterase activity ranged from 121 to 248U/L, with a mean and standard deviation of 186.1±30.68U/L. When comparing plasma cholinesterase activity values in individuals based on the capture site, a significant difference was observed. Establishing reference values for different sea turtle populations is necessary to interpret future sampling results and to allow sea turtles to be used as sentinels of ecosystem health. Future studies are needed to evaluate other populations and the activity of plasma cholinesterase in juvenile marine turtles, in relation to environmental contamination.(AU)

O objetivo desse estudo foi avaliar a atividade enzimática da colinesterase plasmática em tartarugas marinhas da espécie Chelonia mydas em duas populações de acordo com o local de captura, sob ausência e provável influência de efeito antrópico. Foi utilizado um total de 74 animais e posteriormente divididos em dois grupos de acordo com o local de captura. Foram coletadas amostras de sangue de todos os animais capturados e em seguida liberados ao mar no mesmo local. Foi realizada uma análise estatística descritiva dos valores da atividade plasmática de colinesterase do total de animais e análise comparando os valores de acordo com o local de captura. Foram analisadas amostras de plasma heparinizado de animais capturados em dois locais distintos. Os valores da atividade plasmática de colinesterase variaram de 121 a 248U/L, com média e desvio padrão de 186.1±30.7U/L. Quando comparados os valores de atividade plasmática da colinesterase nos indivíduos de acordo com o local de captura, foi observada diferença significativa. O estabelecimento de valores de referência para diferentes populações de tartarugas marinhas são necessários para interpretar os futuros resultados amostrais e permitir que as tartarugas marinhas sejam usadas como sentinelas da saúde do ecossistema. Estudos futuros são necessários para avaliar outras populações e a atividade da colinesterase plasmática de tartarugas marinhas juvenis em relação à contaminação ambiental.(AU)

Atividade da colinesterase plasmática como biomarcador de impacto ambiental em tartarugas verdes (Chelonia mydas) no litoral do Arquipélago de Fernando de Noronha, Pernambuco / Plasma cholinesterase activity as environmental impact biomarker for green turtles (Chelonia mydas) on the coast of Fernando de Noronha, Pernambuco, Brazil

Biomarcadores podem ser usados de forma preditiva, permitindo que sejam tomadas ações de controle antes que ocorram danos ambientais irreversíveis com consequências ecológicas severas, no entanto, espécies sentinelas são necessárias para avaliação desses marcadores. As tartarugas marinhas são consideradas espécies sentinelas quando acometidas por fibropapilomas, sendo sinalizadora do desequilíbrio ambiental marinho nas suas áreas de ocorrência. Com o objetivo de propor a determinação da atividade da colinesterase plasmática em tartarugas verdes (Chelonia mydas) como biomarcador, procedeu-se a determinação da atividade enzimática em animais saudáveis e em localidade de baixo impacto antrópico (Arquipélago de Fernando de Noronha, Pernambuco, Brasil) para servir como referência para comparação com animais capturados em locais de maior impacto antrópico. Ao todo foram analisadas amostras de plasma heparinizado de 35 animais capturados. Todas as amostras analisadas apresentaram alguma atividade enzimática de colinesterase plasmática. Os valores obtidos de colinesterase variaram de 162 a 379 UI/L, com média e desvio padrão de 216,4 ± 51,4 UI/L. Nos estudos de repetibilidade e reprodutibilidade obtiveram-se coeficientes de variação menor que 5% em todas as análises, portanto a metodologia analítica utilizada se mostrou confiável. A longevidade das tartarugas marinhas da espécie C. mydas, o comportamento alimentar, juntamente com o fato de possuirem atividade enzimática detectável podem indicar essa espécie como bioindicadora de exposição a poluentes que influenciam na atividade da colinesterase plasmática.(AU)

Biomarkers can be used in a predictive manner, permitting actions control to be taken before they occur irreversible damage with severe environmental ecological consequences, however, sentinel species are needed to evaluate these markers. Sea turtles affected by fibropapillomas are considered sentinel species, signaling the marine environmental imbalance in their areas of occurrence. With the objective to use the plasma cholinesterase activity in green turtles (Chelonia mydas) as a biomarker, the measurement of enzyme activity was evaluated in healthy animals and in location of low pollution impact (Fernando de Noronha, Pernambuco, Brazil) to serve as a reference for comparison with animals trapped in places with higher pollution impact. Were analyzed heparinized plasma samples from 35 captured animals. All samples analyzed showed enzymatic activity of plasma cholinesterase. The values obtained ranged between 162-379 IU/L, with mean and standard deviation of 216.4 ± 51.4 IU/L. In studies of repeatability and reproducibility were obtained variance coefficients of less than 5% in all analyzes, so the analytical methodology proved successful. The longevity of marine turtles of the species C. mydas, feeding behavior, along with the fact possess detectable enzyme activity may indicate this species as a bioindicator of exposure to pollutants that influence plasma cholinesterase activity.(AU)

Laboratory Abnormalities in Patients with Organophosphorous Poisoning.

Organophosphorous (OP) poisoning is an ever increasing and troublesome situation in the developing countries and is a major health care challenge in the 21st century. Hundred patients who attempted suicide with organophosphates, admitted to the emergency services were included in the study. They were graded (grade 0,1,2,3) according to clinical findings and examined for parameters like RBS, serum urea, serum creatinine, liver function tests, serum amylase, serum cholinesterase and LDH . The APACHE (II) of the cases were determined and correlated with severity of the clinical manifestations. A significant decline in serum cholinesterase (ChE) with increasing grades of intoxication(p <0.05) was observed along with raised levels of random blood sugar, serum urea, creatinine, hepatic enzymes and amylase. APACHE(II) score, showed a significant rise with severity of the degree of intoxication (p<0.001) and a negative correlation with serum cholinesterase. The findings of this study highlighted usefulness of biochemical and clinical indices in the management of organophosphorous poisoning thereby recognizing the complications early and facilitating early management.

Niveles de colinesterasa plasmática en mujeres embarazadas y no embarazadas expuestas a plaguicidas en la zona del Páramo: Mérida-Venezuela / Plasma cholinesterase levels pregnant and non-pregnant women exposed to pesticides Páramo area: Mérida-Venezuela

Se realizó un estudio comparativo y experimental para determinar el nivel plasmático de colinesterasa (con la técnica de Ensayo del reactivo de Ellman) en 51 mujeres embarazadas y 52 no embarazadas, expuestas en forma crónica a insecticidas organofosforados y carbamatos en la zona del páramo merideño y que acudieron a control y/o consulta en el Hospital tipo I de Mucuchíes (Mérida) con la finalidad de establecer la prevalencia de sus efectos en la zona mencionada y la ulterior consecuencia en el producto de la gestación. Los datos se analizaron mediante el software estadístico SPSS versión 15 para Windows. Los resultados mostraron que las pacientes embarazadas con o sin antecedentes patológicos, sus niveles de colinesterasa tienden a disminuir en cierta magnitud, se encontró diferencia estadísticamente significativa entre los niveles de colinesterasa promedio de mujeres embarazadas con antecedentes con respecto a las que no tenían antecedentes, en las mujeres no embarazadas y sin antecedentes que vivían cerca de zonas de cultivo con exposición a plaguicidas el nivel promedio de colinesterasa fue menor al nivel promedio de las mujeres en edad reproductiva no expuestas y una disminución de los valores de colinesterasa promedio en las gestantes menores de 39 años de edad. Se concluyo que las mujeres embarazadas con antecedentes patológicos presentan inhibición de niveles de colinesterasa plasmática con respeto a las no embarazadas y sin antecedentes.

A study was conducted to determine comparative and experimental plasma cholinesterase level (with the assay technique of Ellman's reagent) in 51 pregnant women and 52 non-pregnant, chronically exposed to organophosphate and carbamate wilderness area in Merida and who came to control and / or reference in the type I Mucuchíes Hospital Merida state in order to establish the prevalence of its effects on the area mentioned and further result in the product of gestation. The data were analyzed using SPSS statistical software version 15 for Windows. The results showed that pregnant patients with or without pathological history cholinesterase levels tend to decrease to some extent, statistically significant difference was found between the average cholinesterase levels of pregnant women with a history over which they had no background in nonpregnant women with no history of living near farming areas with pesticide exposure cholinesterase level was lower average to average level of reproductive age women unexposed and decreased average cholinesterase values in pregnant women under 39 years of age. It was concluded that pregnant women with medical history presented inhibition of plasma cholinesterase levels with respect to the non-pregnant and no history.

Study of the correlation between blood cholinesterases activity, urinary dialkyl phosphates, and the frequency of micronucleated polychromatic erythrocytes in rats exposed to disulfoton

Organophosphates (OPs) are widely used as pesticides, and its urinary metabolites as well as the blood cholinesterases (ChEs) activity have been reported as possible biomarkers for the assessment of this pesticide exposure. Moreover, the OPs can induce mutagenesis, and the bone marrow micronucleus test is an efficient way to assess this chromosomal damage. This paper reports a study carried out to verify the correlation among the disulfoton exposure, blood ChEs activity, urinary diethyl thiophosphate (DETP), and diethyl dithiophosphate (DEDTP), as well as micronucleated polychromatic erythrocytes (MNPCEs) frequency. Four groups of rats (n=12) were exposed to disulfoton at 0, 2.8, 4.7, and 6.6 mg kg-1 body weight. The blood ChEs activity, urinary DETP and DEDTP concentrations, and MNPCEs frequency were determined. It was observed that the plasmatic and erythrocytary ChEs activity decreased from 2.9% to 0.5% and from 35.9 to 3.3%, respectively, when the disulfoton dose was increased from 0 to 6.6 mg kg-1 (correlation of 0.99). Urinary DETP and DEDTP concentrations, as well as the MNPCEs frequency, increased from 0 to 6.58 µg mL-1, from 0 to 0.04 µg mL-1, and from 0 to 1.4%, respectively, when the disulfoton dose was increased from 0 to 6.58 mg kg-1 body weight.

Os organofosforados (OPs) são amplamente usados como praguicidas e a atividade da colinesterase sanguínea bem como os metabólitos urinários desses praguicidas têm sido reportados como biomarcadores eficazes para avaliar casos de exposição. Além disso, os OPs podem induzir mutagênese e o teste de micronúcleo de medula óssea é uma boa alternativa para avaliar os danos cromossômicos. Esse artigo reporta um estudo sobre a correlação entre a exposição a dissulfoton, a atividade da colinesterase sanguínea, a excreção urinária de dietil tiofosfato e dietil ditiofosfato e a frequência de micronúcleos em eritrócitos policromáticos. Quatro grupos de ratos (n=12) foram expostos a dissulfotom nas doses de 0, 2,8, 4,7, e 6,6 mg kg-1 de peso corpóreo. A atividade da colinesterase sanguínea as concentrações urinárias de dietil tiofosfato e dietil ditiofosfato e a frequência de micronúcleos foram determinadas. Os resultados demonstraram que as atividades da colinesterase plasmática e eritrocitária diminuíram de 2,9 para 0,5% e de 35,9 para 3,3% , respectivamente, quando a dose de dissulfoton foi aumentada de 0 para 6,6 mg kg-1 (correlação de 0,99). As concentrações urinárias de dietil tiofosfato e dietil ditiofosfato bem como a frequência de micronúcleos aumentaram de 0 a 6,56 µg mL-1, 0 a 0.04 µg mL-1 e de 0 a 1.4%, respectivamente, quando a dose de dissulfotom foi aumentada de 0 a 6,58 mg kg-1.

Evaluation of cholinesterase to differentiate pleural exudates and transudates.

AIMS AND OBJECTIVES: The present study was undertaken to evaluate the usefulness of pleural fluid cholinesterase (PChE) level in pleural fluid and its ratio to serum cholinesterase (P/SChE) in order to differentiate transudates and exudates and to compare their diagnostic efficacy with the Light's criteria. MATERIAL AND METHODS: A total of 110 patients of pleural effusion of diverse etiology were studied. Eighty patients were of exudative pleural effusion of tubercular, malignant or parapneumonic origin and 30 patients were of transudative effusion. Cholinesterase was estimated in the pleural fluid and serum in all the patients. RESULTS: The mean PChE and P/S ChE were significantly higher in exudates as compared to transudates (p < 0.001). P/S ChE was 0.79 +/- 0.45 and 0.14 +/- 0.11 in exudates and transudates, respectively. When a cut-off value of 469 IU/L for PChE was taken for the diagnosis, it was found that 10% of exudates and 2.5% of transudates were misclassified. However percentage of misclassification decreased to 1.25% in exudates and 3.3% in transudates when the cut-off value of 0.24 for P/S ChE ratio was used. Using Light's criteria, a sensitivity of 91.25% and specificity of 90% with positive predictive value (PPV) of 96.05% and negative predictive value (NPV) of 79.42% was observed. However using P/S ChE, the PPV was 98.75% and NPV was 96.67%. CONCLUSIONS: The estimation of PChE and P/SChE ratio had better discriminatory capacity than Light's criteria. It is cost effective and more specific, therefore its routine estimation is recommended.

Health monitoring of farm labourers engaged in MIPC 50 WP field sprays.

Investigations were undertaken to monitor the health status of farm labourers engaged in field sprays of MIPC 50 WP (Hexamicin, Mipcin), a carbamate insecticide on cotton crop, as per the protocol approved by the Central Insecticide Board The insecticide sprays (0.1%) were undertaken for six hr on three consecutive days on 1.2 hectares of cotton crop per day, using Aspee napsak sprayers. The spray personnel (mixers, loaders and sprayers) with protective clothing did not reveal any alteration in clinical, hematological and blood bio-chemical profile during exposure and post exposure periods. The spray personnel without protective clothing showed only a marginal reduction in their blood cholinesterase activity during the exposure period.

Efecto del tiempo y las condiciones de almacenamiento sobre la actividad de las colinesterasas / Effect of time and the conditions of storage in the activity of the cholinesterases

Se evaluó la estabilidad de la colinesterasa plasmática y la colinesterasa eritrocítica a temperatura ambiente, 4§C y -20 §C, durante un periodo de tres meses. Se determinó que la colinesterasa plasmática es estable en refrigeración y congelación. A temperatura ambiente la actividad de esta enzima disminuye significativamente en una semana. Por su parte la colinesterasa eritrocítica es estable únicamente en refrigeración. El proceso de congelamiento y descongelamiento provoca disminución importante en la actividad de la enzima. En las muestras conservadas por un día a -20 §C, la actividad de la enzima eritrocitaria fue enre 10 y 20 por ciento más baja que la actividad basal. A temperatura ambiente la actividad de esta enzima disminuye en forma importante en una semana. Palabras clave: colinesterasa plasmática, colinesterasa eritrocítica, plaguicidas, organofosforados, carbamatos, toxicología

Modificación de un método para colinesterasa plasmática empleando la reducción del ferricianuro como indicador / Modification in the method for serum cholinesterase in ferricyanide reduction how indicatory

Se describe la modificación de un método para la cuantificación de colinesterasa plasmática mediante monitoreo continuo empleando ferricianuro como indicador. La tiocolina liberada de la propioniltiocolina reduce el ferricianuro a ferrocianuro y la disminición de absorbancia a 405 nm es proporcional a la actividad de la enzima. Las precisiones día a día para muestras con valores de colinesterasa bajos y altos mostraron coeficientes de variación de 3,2 y 1,2 por ciento y en un mismo día de 1,1 y 0,52 por ciento respectivamente. La bilirrubina no presenta una interferencia importante y cada 50mg/dL de hemoglobina producen una interferencia de -4 por ciento. El reactivo de ferricianuro almacenado en botella ámbar es estable por al menos 3 meses a temperatura ambiente y 6 meses a 4-8 §C. Al comparar los resultados con un método que emplea la reacción de Ellman se obtuvo una ecuación de regresión lineal de Y = 1,22 (X) - 732, con un coeficiente de correlación (r) de 0,988 y una desviación estándar sobre la línea de regresión (Sy/x) de 378 U/L. (Rev Cost Cienc Med 1999; 20(1,2): 17-27) PALABRAS CLAVE: Colinesterasa sérica, organofosfatos, carbamatos, pesticidas, métodos colorimétrico, toxicología

Estimation of plasma cholinesterase enzyme activity and phenotypes in Pakistani population

The use of plasma cholinesterase for forensic purpose is no different from the use of other enzymes that are genetically variable. There are two advantages, first of all, it is very stable and it deteriorates slowly so that old malodorous blood samples can be used for phenotyping and also scrapings of dried blood stains, when redesolved in buffer, can be used. The second advantage is that the heterozygote E1uE1a can easily be recognised so that pedigree determinations are usually clear. The frequency of different genes varies with different ethnic populations. The real understanding of the genetics of plasma cholinesterase variations is essential for interpretation of data for forensic purposes. The classical method requires a spectrophotometer to monitor at 240nm wave length. As data from many countries were taken, Pakistani data was not available, so this data will enable us in the genetic determination, for treatment of Organophosphorous poisoning cases, and in case of Suxamethonium sensitive individuals

Effect of soman administration on beta-esterases in blood, liver microsomes and brain regions of rats.

Activities of enzymes cholinesterase (ChE) and carboxylesterase (CaE) were assayed in serum, liver microsomes and three regions of brain, viz; cerebrum, cerebellum and brain stem (with mid brain) in male albino rats at 0.5 and 2 h periods after administration of 1/2 LD 50 dose of soman (0.22 mg/kg) intraperitoneally in olive oil as vehicle. At 0.5 h, in serum, ChE activity declined to 33% of its initial level whereas CaE activity was almost completely inhibited. However, in the liver microsomes at this period, ChE activity was greatly inhibited (18% of initial level) whereas CaE activity was nearly unaffected. At 2 h period, both the enzymes in the serum were almost completely inhibited. In the brain regions (excepting in cerebellum), both the enzymes were nearly similarly inhibited (by 55% to 65% of the initial level) at both the periods. The time related differential response of these two beta-esterases in acute soman intoxication probably occurred in the peripheral tissues like blood and liver but not in the CNS.

Comparación de los niveles de pseudocolinesterasas en poblaciones a riesgo por exposición con organofosforados / Comparison of pseudocholinesterase levels in populations at risk due to organophosphorate exposure

Con el propósito de comparar los niveles de pseudocolinesterasas en un grupo de personas que están en contacto con los organofosforados; se estudiaron 300 personas, separandolas en dos grupos : 100 trabajadores de la Compañía Bananera, que usan dichos productos en sus labores diarias, y han recibido preparación en cuanto a su uso y cuentan con supervisión médica, 100 habitantes de la misma zona que tienen sus casa localizadas entre los bananales, sin contacto directo con organofosforados; y 100 habitaciones de una zona urbana cercana quienes constituyen el grupo control. Al finalizar el estudio se detectó un alto porcentaje del 33 por ciento, de niveles bajos de pseudocolinesterasas en los habitantes de las zonas bananeras, mientras que en los empleados de la compañía Bananera fue el 3 por ciento y en la población control un 1 por ciento. No se observa que el sexo tenga relevancia en los resultados obtenidos. Los resultados obtenidos para los residentes de la zona bananera son relevantes, lo que los lleva a ser un grupo de interés social, tanto por el aspecto educativo como por la supervisión médica que ellos merecen, ya que en su relación indirecta con los organofosforados han resultado afectados.

Vigilancia de las enfermedades provocadas por plaguicidas: La experiencia nicaraguense

In 1984, work designed to expanded cholinesterase screening activities and improve the reporting of pesticide poisonings was initiated in Nicaragua's León-Chinandega region as a pilot project. Using a field tintometric method, 1,960 workers were screened for whole blood cholinesterase. The percentage with low cholinesterase activity levels (50 per cent or less) increased sharply during the peak spraying season. Airfield workers were most affected, though a noteworthy share of certain agricultural workers were also found to have low levels. Workers who used certain kinds of personal protective equipment were significantly less affected (p .05). In addition to these survey findings, six deaths and 396 pesticide-related poisonings were reported in the León-Chinandega region in 1984. This indicated a relatively high rate of 74.6 poisoning cases per 100,000 inhabitants, 84 per cent of them occurring in October-December. Ninety-four percent of the cases reported via questionnaires were occupationally related, small farms being the most affected. Menthyl parathion was implicated in roughly half of these cases , two-thirds of which were due to dermal exposure. Policy recommendations derived from the initial results reported here include reduction of methyl parathion use, installation of closed systems for safer aircraft loading, provisión and use of clothing that protects the skin against exposure

Acetylcholine: a possible neurotransmitter in Setaria cervi.

The total and free acetylcholine (Ach) and cholinesterase (CHE) content of adult Setaria cervi were estimated. The Ach was estimated by bioassay on rectus abdominis muscle of frog and the CHE by measuring the drop in pH following incubation of worm homogenate with Ach chloride. The free and total Ach contents (4.0 +/- 0.57 and 6.0 +/- 0.48 microgram/g wet weight of worms respectively) were as high as found in mammalian brain cortex. The cholinesterase activity was found to be 5.57 +/- 0.6 units/g wet weight of worms. It is possible that there may exist a well developed system responsible for the synthesis, storage, release and destruction of Ach and that Ach may be acting as an excitatory neurohormone in S. cervi.