A biomarker model of sublethal genotoxicity (DNA single-strand breaks and adducts) using the sentinel organism Aporrectodea longa in spiked soil

There is a need to develop risk biomarkers during the remediation of contaminated land. We employed the earthworm, Aporrectodea longa (Ude), to determine whether genotoxicity measures could be applied to this organism's intestinal tissues. Earthworms were added, for 24h or 7 days, to soil samples spiked with benzo[a]pyrene (B[a]P) and/or lindane. After exposure, intestinal tissues (crop/gizzard or intestine) were removed prior to the measurement in disaggregated cells of DNA single-strand breaks (SSBs) by the alkaline comet assay. Damage was quantified by comet tail length (CTL, microm). B[a]P 24-h exposure induced dose-related increases (P<0.0001) in SSBs. Earthworm intestine was significantly (P<0.0001) more susceptible than crop/gizzard to B[a]P and/or lindane. However, both tissues appeared to acquire resistance following 7-day exposure. B[a]P-DNA adducts, measured by (32)P-postlabelling, showed a two-adduct-spot pattern. This preliminary investigation suggests that earthworm tissues may be incorporated into genotoxicity assays to facilitate hazard identification within terrestrial ecosystems.     

Concentrations and distribution of 3,3'-dichlorobenzidine and its congeners in environmental samples from Lake Macatawa

Release of 3,3'-dichlorobenzidine (DCB), an intermediate in dye manufacturing processes, is of environmental concern due to its carcinogenic nature. An 11-year field study has been conducted to elucidate the fate and behavior of DCB and its congeners in the Lake Macatawa (Holland, MI, USA) sediment-water system. The sediments were variable in composition, ranging from sandy sediments with 1-8.7% total organic carbon (OC) to silty-clay sediments with 7.5-20.6% total OC. The pH ranged from 6.3 to 7.4. The findings from this field study confirmed that DCB and its congener, benzidine, are transported over long distances. The concentration of DCB in the water phase was found to be from non-detectable to approximately 1300 times greater than the water quality criteria established for DCB (0.021 microg/l). Its congener, benzidine, which is substantially more toxic for humans than its parent compound has been found up to approximately 12300 times higher levels than the EPA guidelines suggest (0.000086 microg/l). Both DCB and benzidine were found in the sediment phase. DCB was observed at almost 70 mg/kg in a sample collected in 1993. The concentrations in both phases have declined in recent years. To assess the threat of the released pollutants, it is necessary to understand how pollutants tend to behave in various media. Therefore, it is crucial to identify and quantify all sources of the chemical and its congeners in order to meaningfully predict the fate (and transport) of a hydrophobic organic compound (HOC) in the environment.     

Bioaccumulation of perfluoroalkyl acids in dairy cows in a naturally contaminated environment

Beef and dairy products may be important vectors of human exposure to perfluoroalkyl acids (PFAAs), but the understanding of how PFAAs are accumulated and transferred through agricultural food chains is very limited. Here, the bioaccumulation of PFAAs in dairy cows receiving naturally contaminated feed and drinking water was investigated by conducting a mass balance of PFAAs for a herd of dairy cows in a barn on a typical Swedish dairy farm. It was assumed that the cows were able to reach steady state with their dietary intake of PFAAs. Perfluorooctane sulfonic acid (PFOS) and perfluoroalkyl carboxylic acids (PFCAs) with 8 to 12 carbons were detected in cow tissue samples (liver, muscle, and blood) at concentrations up to 130 ng kg^?1. Mass balance calculations demonstrated an agreement between total intake and excretion within a factor of 1.5 and consumption of silage was identified as the dominant intake pathway for all PFAAs. Biomagnification factors (BMFs) were highly tissue and homologue specific. While BMFs of PFOS and PFCAs with 9 and 10 fluorinated carbons in liver ranged from 10 to 20, perfluorooctanoic acid (PFOA) was not biomagnified (BMF?<?1) in any of the investigated tissues. Biotransfer factors (BTFs; defined as the concentration in tissue divided by the total daily intake) were calculated for muscle and milk. Log BTFs ranged from ?1.95 to ?1.15 day kg^?1 with the highest BTF observed for PFOS in muscle. Overall, the results of this study suggest that long-chain PFAAs have a relatively high potential for transfer to milk and beef from the diet of dairy cows. However, a low input of PFAAs to terrestrial systems via atmospheric deposition and low bioavailability of PFAAs in soil limits the amount of PFAAs that enter terrestrial agricultural food chains in background contaminated environments and makes this pathway less important than aquatic exposure pathways. The BTFs estimated here provide a useful tool for predicting human exposure to PFAAs via milk and beef under different contamination scenarios.     

The use of radioactive isotopes to measure the transfer of materials in aquatic food chains

Radioisotopes have been misused extensively by ecologists in transfer studies within food chains. Unless it is known that no recycling of isotope has occurred during the experiment, the assumption of linear uptake when in fact the system is not linear, even over short periods, can lead to significant errors in the estimation of ingestion or feeding. If recycling occurs, at least a 3 or 4-compartment, hydraulic-type model is necessary to even approximate the complicated kinetics of isotopic transfer in a simple aquatic feeding experiment. In any event, it is essential to follow the uptake or loss of an isotope (change in specific activity) as a function of time in at least 1 compartment before deciding on an appropriate model. If experiments are designed so that the maximum number of rate processes are summed or integrated by the animal, the kinetics can be considerably simplified. If the food supply is uniformly labelled, the rate of change of tracer can be used to give a rate of ingestion (grazing). If the predator is labelled with a suitable isotope before starting the experiment, the rate of loss of its isotope burden under different experimental conditions can be used to determine respiration or excretion rates, turnover rates, ingestion, and the size and number of major compartments in the transfer system.Bedford Institute Contribution. Woods Hole Oceanographic Institution Contribution No. 2690.     

Cadmium stress alters gene expression of DNA mismatch repair related genes in Arabidopsis seedlings

Cadmium (Cd) is a non essential element, and is a widespread environmental pollutant. Exposure to Cd can result in a variety of adverse health effects in plant and humans. In the current study, Arabidopsis seedlings were used as a bio-indicator of Cd pollution. Seedlings were grown on MS media containing 0-6.0 mg L(-1) Cd for 18 days, and the gene expression patterns were used to link increased Cd exposure with progressive biological effects. Reduction of total soluble protein content in shoots of the Arabidopsis seedlings occurred with increase in Cd concentrations. For the gene expression patterns, seven genes known to be involved in cell division and DNA mismatch repair (MMR) system were investigated by semi-quantitative RT-PCR, and normalized using 18S rRNA gene expression. Expression of the proliferating cell nuclear antigen 2 (atPCNA 2), MutS 3 homolog (atMSH 3) and MutL1 homolog (atMLH1) genes in shoots of Arabidopsis was strongly induced by exposure to 0.75 mg L(-1) Cd, but were repressed by other Cd concentrations whereas exposure to 0.75-6 mg L(-1) of Cd resulted in a decreased expression of atPCNA1, atMSH 2, 6 and 7 genes independently of any observable biological effects, including survival, fresh weight and chlorophyll level of shoots. This work demonstrated that specific gene expression changes could serve as useful molecular biomarkers indicative of Cd exposure and related biological effects.     

Low paternity in the hornet Vespa crabro indicates that multiple mating by queens is derived in vespine wasps

Queen mating frequency was studied in the European hornet, Vespa crabro, by analyzing four DNA microsatellite loci in 20 workers from each of 14 nests. Queens were found to be predominantly singly mated (9/14), although double (4/14) and triple mating (1/14) also occurred. For most multiply mated queens, paternity was significantly biased with the majority male fathering on average 80% of the female offspring. The population-wide effective mating frequency was therefore low (1.11), and sister-sister relatedness high (0.701 ± 0.023 SE). Low effective mating frequency in Vespa, in combination with data from other vespines, suggests that high paternity frequency is derived in the group. Some problems with the non-detection of fathers, where the queen was not sampled or shared alleles with males, are analyzed. Received: 16 November 1998 / Received in revised form: 29 March 1999 / Accepted: 12 April 1999     

Queen control of egg fertilization in the honey bee

The study investigated the precision with which honey bee queens can control the fertilization of the eggs they lay. Because males and workers are reared in different-sized cells, the honey bee is one of the few Hymenoptera in which it is possible for the experimenter to know which type of egg a queen “intends” to lay. Eggs were collected from both worker and drone (male) cells from four honey bee colonies. Ploidy of the embryo was determined using polymorphic DNA microsatellites. All 169 eggs taken from worker cells were heterozygous at at least one microsatellite locus showing that the egg was fertilized. All 129 eggs taken from drone cells gave a single band at the B124 locus, strongly suggesting haploidy. These data show that honey bee queens have great, and quite possibly complete, ability to control the fertilization of the eggs they lay. Data from the literature suggest that in two species of parasitoid Hymenoptera (Copidosoma floridanum, Colpoclypeus florus) females have great, but not complete, ability to control fertilization. Received: 23 December 1997 / Accepted after revision: 17 May 1998     

Laboratory leaching studies of oryzalin and diuron through three undisturbed vineyard soil columns

The leaching of diuron and oryzalin through undisturbed soil columns was studied in the laboratory using three vineyard soils from Vosne-Romanée (Burgundy): a rendosol, a calcosol and a vegetated calcosol. After 845 mm of simulated rainfall in 15 days, soil leachates contained higher amounts of diuron (3.2%, 11.8% and 18.8% of applied diuron, respectively) than oryzalin (0.2%, 4.9%, 3.7%, respectively). A greater proportion of soil extractable residues was obtained for diuron (42.5%, 26.8% and 32.2%, respectively) than for oryzalin (14.7%, 12% and 15.5%, respectively). The greater mobility of diuron might be related to its higher water solubility (36.4 mgl(-1) compared with 2.6 mgl(-1) for oryzalin) and smaller adsorption coefficient (400 lkg(-1), compared with 700-1100 lkg(-1) for oryzalin). The mobility of the two herbicides was greater in the two calcosols than in the rendosol, not only due to different organic carbon contents but also different soil textures and structures.