Altered hepatic cytochrome P450 expression in cats after chronic exposure to decabromodiphenyl ether (BDE-209).

The knowledge of cytochrome P450 (CYP) expression involved in chemical exposure are necessary in clinical applications for the medication and prediction of adverse effects. The aim of this study was to evaluate the mRNA expression of CYP1-CYP3 families in cats exposed to BDE-209 for one year. All selected CYP isoforms showed no significant difference in mRNA expressions between control and exposure groups, however, CYP3A12 and CYP3A131 revealed tend to be two times higher in the exposure group compared to control group. The present results indicate that the chronic exposure of BDE209 could not alter CYP expression in the liver of cats. This result considered caused by the deficiency of CYP2B subfamily which is major metabolism enzyme of polybrominated diphenyl ethers (PBDEs) in cat.

Toxic effects of pyrethroids in tadpoles of Physalaemus gracilis (Anura: Leptodactylidae).

Pyrethroid insecticides are one of the most commonly used pesticide groups, but these compounds have brought risks to non-target species, such as amphibians. This study evaluated the toxicological effects (mortality, swimming activity and oral morphology) caused to a South American species of anuran amphibian, Physalaemus gracilis, exposed to the pyrethroids cypermethrin and deltamethrin. Total spawnings of this anuran were collected in the natural environment and transported to the laboratory where they were kept under controlled conditions. Chronic assays were defined between 0.1 and 0.01 mg L-1 of cypermethrin, and 0.009 and 0.001 mg L-1 of deltamethrin. For cypermethrin, a further chronic toxicity test was performed at 0.05 and 2.0 mg L-1, with hatchlings at stages S.20-S.25. Cypermethrin and deltamethrin were lethal enough to kill over 70% of exposed tadpoles in 1 week at concentrations that can be found in nature (0.01-0.1 mg L-1). The exposure effects also influenced swimming activity and caused changes in oral morphology, which would make it difficult for the animals to survive in their natural habitat. Both pyrethroids presented a risk for P. gracilis, so they should be re-evaluated for non-target wild species.

Allyl-isothiocyanate ameliorates the pre-neoplastic changes induced by the fern Dryopteris nigropalaceae on experimental feeding in Guinea pigs.

Enzootic bovine haematuria, caused by long-term ingestion of ferns, is a chronic disease of hill cattle characterized by neoplastic lesions in the urinary bladder. Objectives of this study were to investigate the toxicity potential of long-term feeding of the fern Dryopteris nigropalaceae and effect of allyl isothiocyanate (AITC) to ameliorate fern toxicity and the associated pathological changes. The LC-MS analysis of the fern showed presence of ptaquiloside (4.5 ±â€¯1.0 µg/g) and pterosin B (39 ±â€¯9.1 µg/g). Groups of animals were fed dried fern powder at the dose of 20% w/w in normal feed and treated with and without AITC at graded doses. Long term feeding of fern induced inflammatory and pre-neoplastic lesions in urinary bladder. The important lesions included cystitis, squamous metaplasia and high-grade dysplasia. Urothelium showed positive immunoreactions for nuclear expression of H-ras and p53. However, no mutation suggestive of neoplastic change was observed on partial mRNA sequences analyses of exon 2 of H-ras and 5 or 7&8 of p53 genes. Strikingly, AITC showed dose-dependent amelioration of pre-neoplastic changes in fern-fed animals. In conclusion, AITC is shown to limit pre-neoplastic changes caused by D. nigropalaceae feeding in guinea pigs.

Chronic effects of nitrogenous compounds on survival and growth of juvenile pink shrimp / Efeito crônico de compostos nitrogenados sobre a sobrevivência e crescimento de juvenis de camarão rosa

Abstract In response to growing worldwide market demand, intensive shrimp farming, based on high feed, has developed over the past decade. The nitrogenous compounds mainly generated by animal excretion can cause deterioration of water quality and produce chronic or even acute toxicity to aquatic animals. As prevention, theoretical safety levels have been estimated from acute toxicity tests and they are traditionally used to prevent toxic effects on biota. However, are those concentrations of nitrogenous compounds really safe to Farfantepenaeus paulensis? The current study aimed to investigate the lethal and sublethal effects of ammonia, nitrite and nitrate to juvenile F. paulensis based on safety levels. Each experiment was performed independently in 100 L tanks for 30 days. The survival rates and wet weight of all shrimps were recorded every 10 days. The concentrations tested for ammonia, nitrite and nitrate were respectively: treatment "T1/4", a quarter of the safety level (0.91 mg/L TA-N, 2.55 mg/L NO2--N and 80.7 mg/L NO3--N); treatment "TSL", the safety level (3.65 mg/L TA-N, 10.2 mg/L NO2--N and 323 mg/L NO3--N); and treatment "T2X", twice the safety level (7.30 mg/L TA-N, 20.4 mg/L NO2--N and 646 mg/L NO3--N). For F. paulensis cultivation, the real safety level for nitrite was estimated to be 2.55 mg/L NO2--N. For ammonia and nitrate, the recommended concentrations were <0.91 mg/L TA-N corresponding to 0.045 mg/L NH3-N and <80.7 mg/L NO3--N, respectively.

Resumo Em resposta à crescente demanda do mercado mundial, a carcinicultura intensiva tem se desenvolvido ao longo da última década. Os compostos nitrogenados gerados principalmente pela excreção dos animais podem causar a deterioração da qualidade da água e produzir toxicidade crônica ou mesmo aguda para os animais cultivados. Como prevenção, os níveis de segurança teóricos são estimados a partir de testes de toxicidade aguda e são tradicionalmente usados para evitar efeitos tóxicos sobre a biota. No entanto, as estimativas das concentrações dos compostos nitrogenados são realmente seguras para Farfantepenaeus paulensis? O presente estudo teve como objetivo investigar os efeitos letais e subletais da amônia, nitrito e nitrato em juvenis de camarão marinho F. paulensis com base em níveis de segurança. Cada experimento foi realizado de forma independente em tanques com capacidade de 100 L durante 30 dias. As taxas de sobrevivência e peso úmido de todos os camarões foram registrados a cada 10 dias. As concentrações testadas para amônia, nitrito e nitrato foram respectivamente: "T1/4", um quarto do nível de segurança (0,91 mg/L N-AT, 2,55 mg/L de N-NO2- e 80,7 mg/L N-NO3-); "TSL", nível de segurança (3,65 mg/L N-AT, 10,2 mg/L de N-NO2- e 323 mg/L N-NO3-); e "T2X", duas vezes o nível de segurança (7,30 mg/L N-AT, 20,4 mg/L de N-NO2- e 646 mg/L de N-NO3-). Para a criação de F. paulensis, o nível de segurança real para nitrito foi estimado em 2,55 mg/L N-NO2-. Para amônia e nitrato, concentrações recomendadas foram: <0,91 mg/L N-AT correspondente a 0,045 mg/L N-NH3 e <80,7 mg/L N-NO3-, respectivamente.

Chronic effects of nitrogenous compounds on survival and growth of juvenile pink shrimp.

In response to growing worldwide market demand, intensive shrimp farming, based on high feed, has developed over the past decade. The nitrogenous compounds mainly generated by animal excretion can cause deterioration of water quality and produce chronic or even acute toxicity to aquatic animals. As prevention, theoretical safety levels have been estimated from acute toxicity tests and they are traditionally used to prevent toxic effects on biota. However, are those concentrations of nitrogenous compounds really safe to Farfantepenaeus paulensis? The current study aimed to investigate the lethal and sublethal effects of ammonia, nitrite and nitrate to juvenile F. paulensis based on safety levels. Each experiment was performed independently in 100 L tanks for 30 days. The survival rates and wet weight of all shrimps were recorded every 10 days. The concentrations tested for ammonia, nitrite and nitrate were respectively: treatment "T1/4", a quarter of the safety level (0.91 mg/L TA-N, 2.55 mg/L NO2--N and 80.7 mg/L NO3--N); treatment "TSL", the safety level (3.65 mg/L TA-N, 10.2 mg/L NO2--N and 323 mg/L NO3--N); and treatment "T2X", twice the safety level (7.30 mg/L TA-N, 20.4 mg/L NO2--N and 646 mg/L NO3--N). For F. paulensis cultivation, the real safety level for nitrite was estimated to be 2.55 mg/L NO2--N. For ammonia and nitrate, the recommended concentrations were <0.91 mg/L TA-N corresponding to 0.045 mg/L NH3-N and <80.7 mg/L NO3--N, respectively.

Detection and measurement of carbohydrate deficient transferrin in serum using immuno-capture mass spectrometry: diagnostic applications for annual ryegrass toxicity and corynetoxin exposure.

The neurological livestock disease annual ryegrass toxicity (ARGT) is caused by the ingestion of the naturally occurring glycolipid toxins - the corynetoxins. Corynetoxins also threaten human health as potential contaminants of the food supply. Presently, there are no routine diagnostic tests for corynetoxins-exposure in humans or livestock. Chronic ingestion of corynetoxins has been modeled in rats exposed to dietary tunicamycins for 12 months and carbohydrate deficient transferrin (CDT) has been previously identified as a candidate disease biomarker. Here, the technique of immuno-capture mass spectrometry (icMS) was used to evaluate serum levels of CDT, discriminating between control and tunicamycins-exposed rats with 85% accuracy. The icMS approach is based on the combination of specific transferrin enrichment with functionalized magnetic beads and automated matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). With no other clinically-relevant diagnostic tests available icMS could be readily adapted for high-throughput clinical assessment of corynetoxins-exposure in humans or livestock.

Molecular and biochemical biomarkers in environmental monitoring: a comparison of biotransformation and antioxidant defense systems in multiple tissues.

The cytochrome P450 pathway and antioxidant responses are known for their responsiveness to environmental pollutants and are frequently used as biomarkers at the transcriptional, translational and catalytic levels. Although molecular responses are often assumed to reflect similar changes in enzyme function, several factors can influence intracellular effects, including mRNA stability and protein turnover, signal sensing and transduction, post-translational modifications of proteins, and multiple mode of action of chemicals in complex mixtures. The aim of this study was to use experimental data for a general discussion on the importance of mechanisms modulating transcriptional and catalytic responses of these pathways, and the resulting implications for environmental monitoring. The European eel Anguilla anguilla was selected as fish model to compare the effects of polluted sediments on gene expression and functional levels of cytochrome P450, glutathione S-transferases, UDP-glucoronosyl transferases, catalase, glutathione peroxidases, superoxide dismutase, glutathione, glutathione reductase, glucose 6-phosphate dehydrogenase and γ-glutamylcysteine ligase in the liver and gills. The overall results confirmed significant changes in gene transcription related to biotransformation and oxyradical metabolism, but also supported the evidence of a frequent dissociation between mRNA expression and protein activity. More similar trends of variations and exposure-dependent relationships was observed in the liver for transcriptional and catalytic responses of those pathways closely regulated by specific interactions between substrate, transcription factors, gene and metabolizing protein (i.e. phase I and phase II). On the other hand, the lower metabolism and the cellular machinery of gill cells may prevent elevated transcriptional responsiveness to be translated to an adequate functional response of a protein. Relationships between transcriptional and catalytic effects were often inconsistent for antioxidant responses confirming the complexity of interactions between exposure to chemical pollutants and regulation of oxidative stress responses. Oxidative stress responses may not necessarily be associated with transcriptional variations of genes, but rather with post-translational modifications of proteins. These mechanisms are just beginning to be revealed in marine organisms, but their characterization will be fundamental for better understanding of the implications of variations in gene expressions according to system, tissue, intensity and duration of exposure.

Fuel oil-induced adrenal hypertrophy in ranch mink (Mustela vison): effects of sex, fuel oil weathering, and response to adrenocorticotropic hormone.

Environmental contamination by petroleum hydrocarbons from anthropogenic sources can be a cause of stress for free-ranging wildlife. The response of wildlife to chemical contaminants requires that the hypothalamic-pituitary-adrenal (HPA) axis be precisely regulated to allow for proper glucocorticoid-mediated adaptive responses. Chronic oral exposure to low concentrations of bunker C fuel oil causes the development of adrenal hypertrophy in male ranch mink (Mustela vison) without increasing serum or fecal glucocorticoid concentrations. This hypertrophy is an adaptive response to fuel oil-induced adrenal insufficiency. To determine if the same phenomenon occurs in female mink or male mink exposed to artificially weathered fuel oil, female mink were fed 0 ppm (mineral oil) or 420 ppm fuel oil and male mink were exposed to 0 ppm, 420 ppm fuel oil, or 480 ppm artificially weathered fuel oil in the diet for 60-62 days. At the end of the exposure, serum glucocorticoid concentrations were assayed along with body and organ weight measurements. Fecal glucocorticoid concentrations were assayed at time points throughout the exposure. Male mink fed fuel oil or weathered fuel oil and female mink fed fuel oil had adrenal enlargement without any significant increases in the serum or fecal concentration of glucocorticoids, which is consistent with fuel oil-induced adrenal insufficiency. To address the physiological consequences of adrenal insufficiency, fuel oil-exposed male mink were administered an adrenocorticotropic hormone (ACTH) stimulation test. Fuel oil-exposed animals had a smaller incremental increase in serum glucocorticoid concentration after ACTH challenge compared to control animals. Our findings provide further evidence that the HPA axis of fuel oil-exposed animals is compromised and, therefore, not able to respond appropriately to the diverse stressors found in the environment.

Bioaccumulation and toxicity of 4-nonylphenol (4-NP) and 4-(2-dodecyl)-benzene sulfonate (LAS) in Lumbriculus variegatus (Oligochaeta) and Chironomus riparius (Insecta).

The discharge of surfactants, such as 4-nonylphenol (4-NP) and linear alkylbenzene sulfonates (LAS), into water bodies leads to accumulation of the chemicals in the sediments and may thus pose a problem to benthic organisms. To study the bioaccumulation of surfactants, Oligochaeta worm Lumbriculus variegatus was exposed to sediment-spiked, [14C]-labeled 4-NP and 4-(2-dodecyl)-benzene sulfonate (C12-LAS) in three different sediments (S1-S3). The sediments were characterized for organic carbon (OC) content and particle size distribution. The acute toxicity was examined by exposing L. variegatus and three to four instar Chironomus riparius (Insecta) larvae in water-only exposure to 4-NP and LAS at different concentrations. After 48-h exposure, lethal water concentrations (LC50) and lethal body residues (LBR50) were estimated using liquid scintillation counting. Chronic toxicity was evaluated in two different sediments by exposing first instar C. riparius larvae to sediment-spiked chemicals at different concentrations. After 10 days, the sublethal effects of surfactants were observed by measuring wet weight and head capsule length. Finally, another 10-day test was set up in order to measure the LAS body residues associated with sublethal effects in C. riparius in S2 sediment. The bioaccumulation test revealed that the bioaccumulation of both 4-NP and LAS increased as the sediment organic matter content decreased. It is assumed that the chemical binding to organic material decreased chemical bioavailability. The acute toxicity tests showed that L. variegatus was more tolerant of 4-NP, and C. riparius was more tolerant of LAS when based on water exposure concentration. The LBR-estimates revealed, however, that L. variegatus tolerated clearly higher tissue residues of both chemicals compared with C. riparius. Both chemicals had sublethal effects on C. riparius growth in sediment exposure, reducing larvae wet weight and head capsule size. 4-NP, however, showed an irregular dose-response pattern. The characteristics of the exposure media affected the bioaccumulation potential of both chemicals. Thus, exposure concentrations offered no prediction of body residue, and therefore it is proposed that organism body residue offered a more accurate dose-metric for chemical exposure than the chemical concentration of the environment.

Early occurrence of spontaneous tumors in CD-1 mice and Sprague-Dawley rats.

It is sometimes difficult to assess the relevance of tumors that occur in treated animals in short-term studies. This report is intended to establish a general profile of tumor occurrence in young control CD-1 mice and Sprague-Dawley rats. Data from 20 rat and 20 mouse carcinogenicity studies conducted between 1990 and 2002 at Huntingdon Life Sciences, UK. were collected and evaluated. The route of administration was either dietary oral gavage, and the analysis was confined to sporadic deaths (decedents) in control groups occurring during the first 50 weeks of study. In addition, tumor occurrence between 50-80 weeks were compared. In mice, the most common tumor was lymphoma, followed by bronchiolo-alveolar adenoma. In rats, the most common tumor was adenoma of the pituitary gland, followed by mammary fibroadenoma, and adenocarcinoma. When studies of up to 50 weeks, between 50 and 80 weeks, and at 2-year termination were compared, there was no great difference in tumor occurrence except in male rats, in which the most common tumor up to 50 weeks on study was lymphoma, whereas the most common tumor between 50-80 weeks and at 2 years was pituitary adenoma.

Carcinogenic susceptibility to N-bis(2-hydroxypropyl)nitrosamine (DHPN) in rasH2 mice.

To evaluate the susceptibility of rasH2 mice to N-bis(2-hydroxypropyl)nitrosamine (DHPN), a potent carcinogen targeting the lung, liver, thyroid, and kidney, male, 6-week old, rasH2 mice and wild-type littermates (non-Tg mice) were given DHPN in drinking water at 0, 20 or 200 ppm, and 0 or 200 ppm, respectively, for 26 weeks. The experiment using rasH2 mice given 200 ppm DHPN and non-Tg mice given 200 and 0 ppm DHPN was completed at 20 weeks, since mortality in these groups was remarkably increased due to hemangiosarcomas of the liver. Histologically, tumors developed in the lung and liver in both rasH2 and non-Tg mice treated with DHPN. In addition, proliferative lesions were observed in the forestomach, urethra, and excretory duct of salivary glands in rasH2 mice given 200 ppm DHPN. RT-PCR analysis showed no marked difference in expression of mRNAs for the transgene and the endogenous mouse ras gene between the whole lung tissue containing a neoplasm and normal lung tissue. Our results suggest that rasH2 mice are highly susceptible to DHPN, the target organs including the forestomach, salivary gland and urethra, which have not been found to develop tumors in previous long-term carcinogenicity studies of DHPN in rats and mice.

Results of chronic dietary toxicity studies of high viscosity (P70H and P100H) white mineral oils in Fischer 344 rats.

Two-year dietary studies were conducted to determine the chronic toxicity and its reversibility, and the carcinogenicity of P70(H) and P100(H) white mineral oils in Fischer-344 rats (F-344). The studies were identical in design and followed the Organization for Economic Cooperation and Development, Guidelines for Testing Chemicals, Guideline 453, 1981. Additional endpoints evaluated were: (1) extent of mineral hydrocarbon deposition in liver, kidneys, mesenteric lymph nodes, and spleen of female rats at 3, 6, 12, 18 and 24 months, and (2) reversibility of effects following cessation of exposure. Dietary concentration were 60, 120, 240, and 1,200 mg/kg/day, adjusted periodically to account for bodyweight changes. Study results were consistent with preceding subchronic studies. No treatment-related mortality, neoplastic lesions, or changes in clinical health, hematology, serum chemistry, or urine chemistry were evident in any group administered either white oil. Statistically significant higher food consumption was noted in the 1,200 mg/kg group males and females exposed to either white oil and statistically significant higher body weights were noted in the 1,200-mg/kg males during the latter portion of the P100(H) study. Higher mesenteric lymph node weights were accompanied by increased severity of infiltrating histiocytes. This occurred to a greater extent with the P70(H) than the P100(H) oil. No other histopathology of significance was observed. Mineral hydrocarbons were detected in the liver following exposure to either oil. Maximal concentrations of mineral hydrocarbons in the liver were similar with both oils but occurred more rapidly with the P70(H) oil. Liver mineral hydrocarbon content returned to near-background levels during the reversibility phase. In conclusion, lifetime exposer of F344 rats to P70(H) and P100(H) white oils resulted in only minimal findings and with no consequence to clinical health. Thus, under the conditions of these studies, the No Observable Adverse Effect Level (NOAEL) for these studies was considered to be 1,200 mg/kg/day.

Bone neoplasms in F344 rats given teriparatide [rhPTH(1-34)] are dependent on duration of treatment and dose.

A long-term study was conducted in female F344 rats to determine the relative importance of dose, treatment duration, and age at initiation of treatment on the incidence of teriparatide [rhPTH[1-34)]-induced bone proliferative lesions. Treatment groups consisted of different combinations of dose (0, 5, or 30 microg/kg/d), treatment duration (6, 20, or 24 months) and age at initiation of treatment (2 or 6 months of age). The primary endpoints were the incidence of bone neoplasms and effects on bone mass and structure as evaluated by quantitative computed tomography and histomorphometery. Significant increases in the incidence of bone tumors (osteoma, osteoblastoma, and osteosarcoma) occurred in rats treated with 30 microg/kg for 20 or 24 months. No neoplasms were found when the 5 microg/kg treatment was initiated at 6 months of age and continued for either 6 or 20 months (up to 70% of life span). This treatment regimen defined a "no-effect" dose for neoplasm formation that nevertheless resulted in substantial increases in bone mass. These results demonstrate that treatment duration and administered dose are the most important factors in the teriparatide-induced bone tumors in rats.

An alternative perspective: a critical evaluation of the Waddell threshold extrapolation model in chemical carcinogenesis.

In a recent Perspective article (Toxicologic Pathology 31: 260-262, 2003) Waddell asserts that he has developed a log linear extrapolation model that can demonstrate a threshold and resolve for once and for all the uncertainies associated with low dose cancer risk extrapolation. However, his method essentially forces, rather than demonstrates, a threshold, and has many serious flaws that result in significant under-estimation of low dose risk. It would be a serious mistake for the scientific community to adopt Waddell's log linear extrapolation model for chemical carcinogenesis risk assessment.