Chronic pesticide poisoning from persistent low-dose exposures in Ecuadorean floriculture workers: toward validating a low-cost test battery

Chronic pesticide poisoning is difficult to detect. We sought to develop a low-cost test battery for settings such as Ecuador’s floriculture industry. First we had to develop a case definition; as with all occupational diseases a case had to have both sufficient effective dose and associated health effects. For the former, using canonical discriminant analysis, we found that adding measures of protection and overall environmental stressors to occupational category and duration of exposure was useful. For the latter, factor analysis suggested three distinct manifestations of pesticide poisoning. We then determined sensitivity and specificity of various combinations of symptoms and simple neurotoxicity tests from the Pentox questionnaire, and found that doing so increased sensitivity and specificity compared to use of acethylcholinesterase alone – the current screening standard. While sensitivity and specificity varied with different case definitions, our results support the development of a low-cost test battery for screening in such settings.

Chronic toxicity of ibuprofen to Daphnia magna: Effects on life history traits and population dynamics

The non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) is a widely used pharmaceutical that can be found in several freshwater ecosystems. Acute toxicity studies with Daphnia magna suggest that the 48 h EC50 (immobilisation) is 10-100 mg IB l(-1). However, there are currently no chronic IB toxicity dataon arthropod populations, and the aquatic life impacts of such analgesic drugs are still undefined. We performed a 14-day exposure of D. magna to IB as a model compound (concentration range: 0, 20, 40 and 80 mg IB l(-1)) measuring chronic effects on life history traits and population performance. Population growth rate was significantly reduced at all IB concentrations, although survival was only affected at 80 mg IB l(-1). Reproduction, however, was affected at lower concentrations of IB (14-day EC50 of 13.4 mg IB l(-1)), and was completely inhibited at the highest test concentration. The results from this study indicate that the long-term crustacean population consequences of a chronic IB exposure at environmentally realistic concentrations (ng l(-1) to mu g l(-1)) would most likely be of minor importance. We discuss our results in relation to recent genomic studies, which suggest that the potential mechanism of toxicity in Daphnia is similar to the mode of action in mammals, where IB inhibits eicosanoid biosynthesis. (C) 2007 Elsevier Ireland Ltd. All rights reserved.

Sublethal neonicotinoid insecticide exposure reduces solitary bee reproductive success

1. Pollinating insects provide crucial and economically important ecosystem services to crops and wild plants, but pollinators, particularly bees, are globally declining as a result of various driving factors, including the prevalent use of pesticides for crop protection. Sublethal pesticide exposure negatively impacts numerous pollinator lifehistory traits, but its influence on reproductive success remains largely unknown. Such information is pivotal, however, to our understanding of the long-term effects on population dynamics. 2 We investigated the influence of field-realistic trace residues of the routinely used neonicotinoid insecticides thiamethoxam and clothianidin in nectar substitutes on the entire life-time fitness performance of the red mason bee Osmia bicornis. 3 We show that chronic, dietary neonicotinoid exposure has severe detrimental effects on solitary bee reproductive output. Neonicotinoids did not affect adult bee mortality; however, monitoring of fully controlled experimental populations revealed that sublethal exposure resulted in almost 50% reduced total offspring production and a significantly male-biased offspring sex ratio. 4 Our data add to the accumulating evidence indicating that sublethal neonicotinoid effects on non-Apis pollinators are expressed most strongly in a rather complex, fitness-related context. Consequently, to fully mitigate long-term impacts on pollinator population dynamics, present pesticide risk assessments need to be expanded to include whole life-cycle fitness estimates, as demonstrated in the present study using O. bicornis as a model.

Prolonged exposure to palmitate impairs fatty acid oxidation despite activation of AMP-activated protein kinase in skeletal muscle cells

The aim of this study was to investigate the chronic effects of palmitate on fatty acid (FA) oxidation, AMPK/ACC phosphorylation/activation, intracellular lipid accumulation, and the molecular Mechanisms involved in these processes in skeletal muscle cells. Exposure of L6 myotubes for 8 h to 200, 400, 600, and 800 mu M of palmitate did rot affect cel viability but significantly reduced FA oxidation by similar to 26.5%, similar to 43.5%, similar to 50%, and similar to 47%, respectively. Interestingly, this occurred despite significant increases in AMPK (similar to 2.5-fold) and ACC (similar to 3-fold) phosphorylation and in malonyl-CoA decarboxylase activity (similar to 38-60%). Low concentrations of palmitate (50-100 mu M) caused an increase (similar to 30%) in CPT-I activity. However, as the concentration of palmitate increased, CPT-I activity decreased by similar to 32% after exposure for 8 h to 800 mu M of palmitate. Although FA uptake was reduced (similar to 35%) in cells exposed to increasing, palmitate concentrations, intracellular lipid accumulation increased in a dose-dependent manner, reaching values similar to 2.3-, similar to 3-, and 4-fold higher than control in muscle cells exposed to 400, 600, and 800 mu M palmitate, respectively. Interestingly, myotubes exposed to 400 mu M of palmitate for 1h increased basal glucose uptake and glycogen synthesis by similar to 40%. However, as time of incubation in the presence of palmitate progressed from 1 to 8h, these increases were abolished and a time-dependent inhibition of insulin-stimulated glucose uptake (similar to 65%) and glycogen synthesis (30%) was observed in myotubes. These findings may help explain the dysfunctional adaptations that occur in glucose and FA Metabolism in skeletal muscle under conditions of chronically elevated circulating levels of non-esterified FAs. Such as in obesity and Type 2 Diabetes.

Toll-like receptors-related genes in kidney transplant patients with chronic allograft nephropathy and acute rejection

Introduction: Toll-like receptors (TLR) comprehend an emerging family of receptors that recognize pathogen-associated molecular patterns and promote the activation of leukocytes. Surgical trauma and ischemia-reperfusion injury are likely to provide exposure to endogenous ligands for TLR in virtually all kidney transplant recipients. Methods: Macroarray (GEArray OHS-018.2 Series-Superarray) analyses of 128 genes involved in TLR signaling pathway were performed in nephrectomy samples of patients with chronic allograft nephropathy (CAN) and acute rejection (AR, vascular and non vascular). The analysis of each membrane was performed by GEArray Expression Analysis Suite 2.0. Results: Macroarray profile identified a gene expression signature that could discriminate CAN and AR. Three genes were significantly expressed between CAN and vascular AR: Pellino 2; IL 8 and UBE2V1. In relation to vascular and non-vascular AR, there were only two genes with statistical significance: IL-6 and IRAK-3. Conclusion: Vascular and non-vascular AR and CAN showed different expression of a few genes in TLR pathway. The analysis of nephrectomy showed that activation of TLR pathway is present in AR and CAN. (C) 2008 Elsevier B.V. All rights reserved.

Chronic contamination assessment integrating biomarkers' responses in transplanted mussels-A seasonal monitoring

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Chronic restraint or variable stresses differently affect the behavior, corticosterone secretion and body weight in rats

Organisms are constantly subjected to stressful stimuli that affect numerous physiological processes and activate the hypothalamo-pituitary-adrenal (HPA) axis, increasing the release of glucocorticoids. Exposure to chronic stress is known to alter basic mechanisms of the stress response. The purpose of the present study was to compare the effect of two different stress paradigms (chronic restraint or variable stress) on behavioral and corticosterone release to a subsequent exposure to stressors. Considering that the HPA axis might respond differently when it is challenged with a novel or a familiar stressor we investigated the changes in the corticosterone levels following the exposure to two stressors: restraint (familiar stress) or forced novelty (novel stress). The changes in the behavioral response were evaluated by measuring the locomotor response to a novel environment. In addition, we examined changes in body, adrenals, and thymus weights in response to the chronic paradigms. Our results showed that exposure to chronic variable stress increased basal plasma corticosterone levels and that both, chronic restraint and variable stresses, promote higher corticosterone levels in response to a novel environment, but not to a challenge restraint stress, as compared to the control (non-stressed) group. Exposure to chronic restraint leads to increased novelty-induced locomotor activity. Furthermore, only the exposure to variable stress reduced body weights. In conclusion, the present results provide additional evidence on how chronic stress affects the organism physiology and point to the importance of the chronic paradigm and challenge stress on the behavioral and hormonal adaptations induced by chronic stress. (c) 2006 Elsevier B.V. All rights reserved.

Effects of chronic stress on nicotine-induced locomotor activity and corticosterone release in adult and adolescent rats

We examined nicotine-induced locomotion and increase in corticosterone plasma levels in adolescent and adult animals exposed to chronic restraint stress. Adolescent [postnatal day (P) 28-37] and adult (P60-67) rats were restrained for 2 hours once daily for 7 days. Three days after the last exposure to stress, the animals were challenged with saline or nicotine (0.4 mg/kg subcutaneously). Nicotine-induced locomotion was recorded in an activity cage. Trunk blood samples were collected in a subset of adolescent and adult rats and plasma corticosterone levels were determined by radioimmunoassay. Exposure to stress did not affect the nicotine-induced locomotor- or corticosterone-activating effects in both ages.

Effects of two chronic stress models on ligature-induced periodontitis in Wistar rats

Objectives: The present study aims at comparing the effects of physical and variable chronic stress on ligature-induced periodontitis in rats.Design: Forty-eight adult Wistar rats were randomly assigned to four groups (n = 12): physical stress, variable stress, positive control and negative control. The models of physical stress were immobilization and immobilization associated with exposure to cold. The models of variable stress were exposure to intermittent light, 24 h isolation, oral cavity examination, crowded environment, smell of blood and noise. After 10 days of physical or variable stress animals underwent experimental induction of periodontal disease in one oral side. Positive control also underwent experimental induction of periodontal disease on the 10th day. Negative control did not receive any type of intervention. At the end of the experimental period (60 days), all animals were euthanized. After routine laboratory processing, images of the histological sections were digitised and submitted to histometric measurement using two parameters: histologic attachment loss and bone loss.Results: Histological attachment loss and bone loss were greater (p < 0.05) in the physical stress group than in the other groups (variable stress, positive and negative control groups). on the non-periodontitis side, these same histological parameters did not significantly differ amongst groups.Conclusions: Physical stress negatively modulated the response pattern to experimentally induced periodontitis in rats. (C) 2011 Elsevier Ltd. All rights reserved.

Oral exposure to methylmercury modifies the prostatic microenvironment in adult rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Decreased endothelium-dependent vasoconstriction to noradrenaline in acute-stressed rats is potentiated by previous chronic stress: Nitric oxide involvement

1. The objective was to determine whether nitric oxide participates in stress adaptive responses. Acute stress (AS) decreased endothelium-dependent vasoconstriction to noradrenaline (NA) in rat aorta [control rat (CR) 3.90+/-0.18, n=22; AS 2.76+/-0.20, n=13; P<0.05].2. Chronic stress exposure previous to AS (CS) potentiated this effect [CS 1.93+/-0.19; n=9; P<0.05 related to CR, P<0.05 related to AS].3. Methylene blue and N-G monomethyl-L-arginine, but not indomethaein, restored the decreased aorta reactivity to NA. 4. No reactivity alteration was observed in aorta without endothelium either in both stress conditions or in the presence of inhibitors. These data show that the nitric oxide participates in stress responses. (C) 1998 Elsevier B.V.

Acclimation of broiler chickens to chronic high environmental temperature

The present study was conducted to evaluate the zootechnical parameters and age related changes in physiological responses of broiler chickens exposed to hot environment from early age onwards. The broiler chickens were exposed to high temperature (30 degreesC) at 15 d of age and maintained to Day 38 or maintained under thermoneutral environment (control).No significant decrease in feed consumption (FC) and body weight (BW) gain was observed in high temperature group after 7 d of exposure, but in the subsequent period, heat exposure lowered BW and FC, compared to control group. However, the weight gain was not significantly changed after 24 d of exposure, and the feed efficiency was not affected throughout the experimental period.The venous pCO(2) pressure was only significantly decreased by temperature after 24 d of heat exposure. The glucose, non-esterified fatty acid (NEFA), triglyceride (TG), glucose, lipid peroxidation (LPO), creatine kinase (CK), and corticosterone were not influenced by the temperature treatment. The significant decrease in uric acid and increase in lactate concentration due to high temperature were observed respectively at 28 and 35 d of age. The concentrations of triiodothyronine (T-3) and thyroxine (T-4) were changed oppositely at 28 d of age, as T-3 was decreased and T-4 was elevated by high temperature. However, the concentration of T-4 in plasma was decreased whereas T-3 was not changed at 38 d of age. The relationships between the blood parameters were changed due to the temperature treatment, suggesting that not only absolute values but also their interrelationships have to be considered when studying the effects of a particular treatment on physiological functioning.These results suggest the growth and physiological responses of broiler chickens, exposed to high temperature from early age onwards, differed at different stages of acclimation. The process of heat acclimation is related to the mode of heat exposure imposed and is not only reflected in the changes in the absolute concentrations, but also in the correlations among the blood indices.

Chronic mild prenatal stress exacerbates the allergen-induced airway inflammation in rats

The effects of chronic mild prenatal stress on leukocyte infiltration into the airways was investigated in rat offspring. The chronic prenatal stress consisted of transitory and variable changes in the rat's living conditions. Offspring at adult age were actively sensitized (day 0) and intratracheally challenged (day 14) with ovalbumin. Bronchoalveolar lavage was performed in the offspring at 48 h after intratracheal challenge with ovalbumin. A significant increase in total leukocyte infiltration was observed in the non-stressed offspring group and this was associated with a marked recruitment of eosinophils without a significant effect on the influx of neutrophils and mononuclear cells. In the prenatal stressed offspring, the counts of both total leukocyte and eosinophils, as well as mononuclear cells, was increased by 50% compared to the non-stressed offspring. We provide here the first experimental evidence that chronic mild unpredictable prenatal stress produces a marked increase in the allergen-induced airway inflammation in the rat offspring.

Effects of Chronic Physical Activity and of Ultrasound Treatment on Bone Consolidation

The objective of this study was to investigate the effects of ultrasound treatment and physical exercise on the velocity of bone consolidation and resistance to deformation. We performed osteotomy in the upper third of the right tibia of rats. Physical training consisted of swimming 1 h per day with a load of 5% b.w. for 30 days. Therapy with medium-intensity ultrasound was applied daily on the damaged area. Wistar rats were divided into the following groups: osteotomized sedentary animals with no ultrasound treatment (1.OSnUS), osteotomized trained animals with no ultrasound treatment (2.OTnUS), osteotomized sedentary animals with ultrasound treatment (3.OSwUS). and osteotomized trained animals with ultrasound treatment (4.OTwUS). The animals were sacrificed for the following analyses: muscle glycogen, serum alkaline phosphatase at the 5th, 10th, 20th, and 30th days, test of maximum resistance to flexion, rupture flexion and mean tibial rigidity at the 30th day. Muscle glycogen was increased at the 20th day: alkaline phosphatase was elevated at the 5th and 20th days in groups 3.OSwUS and 4.OTwUS. and decreased at the 10th day. Groups1.OSnUS and 2.OTnUS did not show significant variations. In the mechanical resistance tests, we noted that ultrasound therapy and the association of physical activity used in the present study showed significant differences in bone resistance and bone rigidity after 30 days of treatment. These facts suggest that ultrasound or physical activity, or their combination may accelerate the process of bone tissue repair.

Ultrastructural and morphometric analysis on the ovary of Wistar rats after chronic ethanol ingestion

In the present study, seventy-two adult rats (Rattus norvegicus albinus) aged three months were used. The animals were divided into two groups (control and alcoholic). The control group received a solid diet (Purina rat chow) and tap water ad libitum. The alcoholic group received the same solid diet and sugar-cane liquid (trade 51, 41° Gay Lussac - GL) diluted 30° GL. At the end or 90, 180 and 270 days of treatment, ten rats of each group were anaesthetized with ethyl ether and sacrificed. The ovaries were collected, fixed, included and submitted to analysis by both light and electron microscopy. The alcoholic group showed increase in the number of corpora lutea at both 180 and 270 days of treatment, atresic follicles at 270 days of treatment, decreased diameter of corpora lutea at 180 and 270 days of treatment, the granulosa layer of the antral follicles at 180 days of treatment, and gradual regression of the theca antral follicles. Furthermore, an increase in diameter and posterior regression of the antral follicle were observed, as well as vacuolation, increased lipid droplets in the granulosa cell at 90 days and in the theca at 180 and 270 days of treatment and gradually in the interstitial cell. The rats showed ovarian alterations after ingestion of alcohol. There was a correlation between exposure time to the drug and the injury observed.