Monoterpene biotransformation by Colletotrichum species.

OBJECTIVE: To investigate the biocatalytic potential of Colletotrichum acutatum and Colletotrichum nymphaeae for monoterpene biotransformation. RESULTS: C. acutatum and C. nymphaeae used limonene, α-pinene, ß-pinene, farnesene, citronellol, linalool, geraniol, perillyl alcohol, and carveol as sole carbon and energy sources. Both species biotransformed limonene and linalool, accumulating limonene-1,2-diol and linalool oxides, respectively. α-Pinene was only biotransformed by C. nymphaeae producing campholenic aldehyde, pinanone and verbenone. The biotransformation of limonene by C. nymphaeae yielded 3.34-4.01 g limonene-1,2-diol l-1, depending on the substrate (R-(+)-limonene, S-(-)-limonene or citrus terpene (an agro-industrial by-product). This is among the highest concentrations already reported for this product. CONCLUSIONS: This is the first report on the biotransformation of these terpenes by Colletotrichum spp. and the biotransformation of limonene to limonene-1,2-diol possibly involves enzymes similar to those found in Grosmannia clavigera.

Biochemical responses, morphometric changes, genotoxic effects and CYP1A expression in the armored catfish Pterygoplichthys anisitsi after 15 days of exposure to mineral diesel and biodiesel.

Despite being considered friendlier to the environment, biodiesel fuel can be harmful to aquatic organisms, especially when combined with petroleum diesel fuel. In this work we evaluated the effects of mineral diesel fuel containing increasing concentrations of biodiesel (5% and 20%, namely B5 and B20) and pure biodiesel (B100), at concentrations of 0.001 and 0.01mLL(-1), after 15 days of exposure, in armored catfish (Pterygoplichtys anisitsi). Toxicity tests were also performed to estimate LC50 values (96h) for each compound. Biotransformation enzymes [ethoxyresorufin-O-deethylase (EROD), and glutathione S-transferase (GST)] as well as oxidative stress markers (superoxide dismutase, SOD, catalase, CAT, glutathione peroxidase, GPx, and the level of lipid peroxidation) were measured in liver and gills after treatment. Genotoxic effects were also accessed in erythrocytes using the comet assay and by evaluating the frequency of micronuclei formation. Further, the mRNA of cytochrome P450 1A (CYP1A) was also measured in liver. Mortality was not observed even exposure to concentrations as high as 6.0mLL(-1). EROD and GST activities were increased after B5 and B20 treatments; however, CYP1A mRNA induction was not observed. SOD and CAT activities were decreased, but GPx was significantly higher for all treatments in gills. There were no significant changes in lipid peroxidation, but genotoxicity markers revealed that all treatments increased comet scores. Fuels B5 and B20 increased micronuclei frequency. Our results indicate that despite being less toxic, biodiesel may cause sublethal alterations in fish that may alter long term health.

A simple R-based function to estimate lethal concentrations.

Every year thousands of new substances are synthetized and marketed. Either by regulatory demands or scientific purposes, these substances should all be tested. The determination of lethal concentration (LC) in a given population after toxicity testing in the laboratory is still the best way to establish and compare toxicity. In this technical note, we propose an R function to estimate LC with improvements on the choice of experimental design and calculation parameters, as well as in editing and spreadsheet loading. Although only LC50 values are considered herein, the same statistical considerations apply to EC50 and ED50 from quantal (dichotomous, binomial, binary) data. A running example is also provided.

Assessment of toxicity of dissolved and microencapsulated biocides for control of the Golden Mussel Limnoperna fortunei.

Biological invasions currently pose major threats to ecosystems worldwide. Invasive bivalves such as the Golden Mussel Limnoperna fortunei can act as 'environmental engineers', altering biogeochemistry, reducing biodiversity, and literally changing the landscape of aquatic environments. The risk that this mussel will invade the Amazon basin is a great concern for environmental authorities, especially because no efficient control methods presently exist. In this study, we tested new microencapsulated chemicals, along with the traditional dissolved chlorine and KCl, as alternatives to control L. fortunei infestation in industrial and water supply plants along rivers. Because these bivalves can close their valves when they sense toxic substances in the water, microencapsulation has improved the effectiveness of the chemicals in controlling L. fortunei, reducing variation in the application and increasing toxicity compared to dissolved chemicals. Microencapsulation should be seriously considered as an alternative to replace hazardous chlorine.

A description of mercury in fishes from the Madeira River Basin, Amazon, Brazil

Over the last 20 years several projects carried on the Madeira River basin in the Amazon produced a great amount data on total Hg concentration in different fish species. In this paper we discuss temporal trends in Hg contamination and its relation to body weight in some of those fishes, showing that even within similar groups, such as carnivorous and non-migratory fish, the interspecies variability in Hg accumulation is considerable.

Vários estudos têm sido desenvolvidos nos últimos 20 anos na bacia do Rio Madeira (Amazônia) com o objetivo de diagnosticar a presença de mercúrio em peixes e compreender o ciclo deste elemento no meio ambiente tropical. Neste artigo são discutidas tendências temporais na concentração de Hg e sua relação com a massa corporal de algumas espécies de peixes com diferentes hábitos alimentares, coletadas no Rio Madeira e no reservatório da hidroelétrica de Samuel, no Rio Jamari, Estado de Rondônia. Foi avaliada uma amostragem de peixes de 14 anos (1987 - 2000) com 86 espécies e um total de 1100 espécimes.

Maternal mercury exposure and neuro-motor development in breastfed infants from Porto Velho (Amazon), Brazil.

Fish is an important item in the diet of Amazonians, and per se is their best single source of essential nutrients. Rapid urbanization and migration are bringing changes in dietary habits of Amazonians. Exposure to fish-Hg during pregnancy and lactation were studied in 100 women and newborns from Porto Velho. Tissue-Hg concentrations and neurodevelopment (Gesell Developmental Schedules) were assessed at birth and at 6 months in exclusively breastfed infants. Maternal mean frequency of fish consumption was low (<2 meals/week; range 0->7 meals/week) compared to Amazonian standards. Women consuming <2 fish meals/week showed less median hair-Hg (3.5 microgg-1) than women that consumed 2 fish meals/week (5.7 microgg). Median total Hg in maternal hair (5.4 microgg-1) was higher than in newborns (1.6 microgg-1). Significant correlation was observed between maternal hair-Hg and infant hair-Hg at birth (r=0.353; p<0.01) and at six months (r=0.510; p<0.01). Placenta-Hg was also significantly correlated to maternal hair-Hg (r=0.321; p<0.01), newborn hair-Hg (r=0.219; p<0.05), maternal blood-Hg (r=0.250; p<0.01) and to umbilical cord-Hg (r=0.857; p<0.01). Most infants (74%) had normal Gesell Schedules but among the 26% showing neuro-motor development delays only six (7%) had multiple (motor, language, and adaptative) delays. The infants with multiple delays were born from mothers with range of hair-Hg comparable to mothers of normally developed infants. Coincidentally, mothers of infants with multiple delays also showed the lowest range of income and level of education. Fish consumption, income, and level of education varied greatly among these breastfeeding urban mothers. It seems that development delays of exclusively breastfed infants are a component of the health inequalities that accompanies socioeconomic disadvantages.

Expression of PXR, CYP3A and MDR1 genes in liver of zebrafish.

The Pregnane X Receptor (PXR) is a nuclear receptor involved in the transcriptional regulation of drug-metabolizing enzymes and transporters. In mammals, many xenobiotics induce the expression of cytochrome P4503A (CYP3A) and the multiple drug resistance 1 (MDR1) genes via the PXR pathway. Little attention has been given to studies about the identification and biological function of PXR homologues in non-mammalian species. Zebrafish is being widely used and accepted as model for toxicological and pharmacological studies to understand the mechanisms of human diseases and identify conserved signaling pathways. The aim of this study was to evaluate the in vivo expression of PXR, CYP3A and MDR1 genes in liver of zebrafish treated with the synthetic steroid pregnenolone 16alpha-carboninitrile (PCN), the antimycotic clotrimazole (CTZ) and the antianginal drug nifedipine (NIF). The liver of fish treated with PCN showed a 1.9-fold induction in the PXR followed by 1.8-fold induction in the CYP3A and 1.6-fold induction in the MDR1 mRNA. CTZ and NIF did not affect statistically the expression of PXR, CYP3A and MDR1. The similar pattern of mRNA expression of PXR, CYP3A and MDR1 genes found in fish treated with different PXR inducers suggests that the intrinsic association between these three genes is conserved in zebrafish.

Bioaccumulation and depuration of Zn and Cd in mangrove oysters (Crassostrea rhizophorae, Guilding, 1828) transplanted to and from a contaminated tropical coastal lagoon.

In order to study Zn and Cd accumulation and depuration, a set of oysters, Crassostrea rhizophorae, were transplanted to a metal contaminated coastal lagoon and another one was harvested there and transplanted to a non-polluted site. C. rhizophorae oysters and Perna perna mussels native from both sites were collected in order to monitor variability of metal concentration in resident populations. After three months exposure, oysters transplanted to the polluted site accumulated fourfold Zn (307-1319 microgg(-1)) without reaching the concentration level of resident oysters (9770 microgg(-1)). Cadmium concentrations had a slight but significant decrease during the same period (1.25-0.54 microgg(-1)). Oysters transplanted to the non-polluted site, showed threefold Zn depuration (6727-2404 microgg(-1)), while Cd had no significant variation (0.90-1.45 microgg(-1)). Results showed that transplanted oysters do not reach heavy metal concentrations in indigenous populations suggesting transplanted organisms would be better used to evaluate bioavailability instead of environmental concentrations.