Ecotoxicology and population genetics: the emergence of "phylogeographic and evolutionary ecotoxicology".

Genetics of ecotoxicology has recently emerged as a priority research field. The advent of polymerase chain reaction and molecular population genetics has made it possible to examine the genetics in even the smallest individuals. Although a potentially powerful technique, current approaches oversimplify the relationship of change in gene frequency to contaminant exposure. Many of these approaches cannot control for random correlation or accessory abiotic factors that impinge on the system tested. Indeed, the gestalt approaches of laboratory exposure or natural field experiments may ignore significant genome-level interactions that are important within a given system. At the very least, these approaches would benefit by a biogeographic survey of genetic variation to understand geographic microevolutionary patterns, or phylogeography, within a species to reduce spurious correlations and erroneous conclusions. Other single locus approaches can be chosen to enhance this approach if genetic/environmental interactions have been characterized for laboratory populations or for other model systems.

Differential survival of three mitochondrial lineages of a marine benthic copepod exposed to a pesticide mixture.

In South Carolina estuaries, the harpacticoid copepod Microarthridion littorale (Poppe 1881) consists of three distinct mitochondrial lineages (liI, liII, and liIII), whose distributions may be partially explained by the presence of toxic contaminants in the sampled habitats. The frequencies of liII and liIII are greatly diminished and sometimes absent in South Carolina contaminated tidal creeks where liI is omnipresent. In this study, representatives of these lineages or haplotype groups were collected from sediments of an estuarine creek containing low to undetectable levels of toxicants and then exposed to a toxic (approximately LC90) aqueous mixture containing an organophosphate (chlorpyrifos) and organochlorine pesticide (DDT, mixed isomers). A comparison was conducted for the frequency of each of the three haplotypes among the survivors of the exposed animals relative to that among the survivors of the control group. The haplotype group with the highest frequency in contaminated SC estuaries (liI) was statistically higher in frequency in survivors of the pesticide-exposed group than in the control group. The two rarer groups (liII and liIII) were less abundant among the survivors of the pesticide-exposed group than the control group. The frequencies of liI, liII, and liIII did not change significantly among the survivors of the control group. The differential survival of the three haplotype groups in the pesticide mixture may be one of the reasons that some haplotype groups are more likely to be found in clean or contaminated tidal creeks on the South Carolina coast.

An efficient DNA extraction method for small metazoans.

The isolation of total nucleic acids from small metazoan taxa is difficult and often leads to an unacceptably large percentage of unsuccessful polymerase chain reaction (PCR) amplifications. Our work with the evolutionary genetics of harpacticoid copepods was an incentive to refine techniques such that consistent amplifications from minute marine organisms were feasible. We describe these modifications and demonstrate their utility for the amplification of multiple loci from single harpacticoid copepods.

Reproductive output of a meiobenthic copepod exposed to sediment-associated fenvalerate.

The benthic harpacticoid copepod, Amphiascus tenuiremis, was cultured through one generation at four concentrations (600, 300, 150, and 0 micrograms/kg) of sediment-associated fenvalerate, a synthetic pyrethroid insecticide, to assess effects on reproductive output and age structure. Two static renewal experiments with four replicates of each treatment were conducted. At the end of 21 days, surviving copepods were sorted and counted into adult males, adult females, copepodites, and nauplii. Fenvalerate was extracted from the sediment and measured by GC-MS. The number of individuals in each life-history stage in fenvalerate treatments was not significantly different from controls, except for increased adult females in fenvalerate in the second experiment. The overall lack of a fenvalerate toxic effect on A. tenuiremis was most likely because fenvalerate tightly binds to sediments and was probably not bioavailable to the copepods. Sediment-associated pesticides with large octanol:water partitioning coefficients (K(ow)'s) such as fenvalerate appear to be less toxic to infaunal copepods than those exhibiting smaller K(ow)'s.

Are Temperature and Photoperiod Necessary Cues for Encystment in the Marine Benthic Harpacticoid Copepod Heteropsyllus nunni Coull?

Heteropsyllus nunni is a marine copepod that builds a cyst and dwells within it during a period of extended diapause. The field abundance of this copepod has been monitored for 10 years, but nothing is known about the cues that induce and terminate encystment. In the laboratory, different photoperiods and temperatures were tested for their effects on encystment and excystment. The photoperiod and temperature cues tested neither induced nor inhibited encystment in H. nunni. Encystment occurred in all treatments, regardless of temperature or photoperiod, suggesting that internal genetic cues, tied to a specific ontogenetic stage, must be the central causal factor. Copepods in the hot treatments encysted and excysted more rapidly than in the cold. Many copepods in the cold treatment encysted (though later than copepods in the hotter treatments), and most were still within the cyst at the end of the 23-week experiment. There were significantly more males within the full cysts than females. A concurrent field study confirmed the known seasonal patterns in the number of encystments relative to the number of free-living forms; i.e., encystment took place in the summer.

Encystment discovered in a marine copepod.

It has been believed that marine crustacea do not encyst as adults. The benthic copepod Heteropsyllus nunni has been found to encyst in intertidal sands in the summer; free-living forms are abundant in the winter. A unique platelike structure on the cephalothorax is thought to be the site of cyst formation.