Genetic study of the extent and consequences of sexual and asexual reproduction in the deep-sea epizoic anemones Amphianthus inornata and Kadosactis commensalis (Cnidaria: Anthozoa)

The anemone Amphianthus inornata is found at bathyal depths living on colonies of the gorgonian Acanella arbuscula. Previous studies of the morphology and reproductive stage of this anemone, during different times of the year, have indicated that it reproduces sexually on a seasonal basis. A small proportion of the study population were also reported to be undergoing asexual reproduction by fission. The anemone Kadosactis commensalis is also bathyal, but lives mainly on the holothurian Paroriza prouhoi. Previous morphological studies have indicated that K. commensalis is a protandrous hermaphrodite that exhibits non-seasonal sexual reproduction only. In the present study, allozyme electrophoresis was used to examine the prevalence and genetic consequences of asexual reproduction in a population of Amphianthus inornata from 2 200 m in the Rockall Trough, North Atlantic Ocean. Genetic evidence, from five randomly selected polymorphic enzyme loci, for asexual reproduction in this species was weak. Exact tests indicated that genotype frequencies did not differ significantly from those expected under Hardy–Weinberg equilibrium. F _IS (correlation of homologous alleles with reference to local population, assuming random mating) values also did not differ significantly from zero, and observed heterozygosity (H _o =0.446) and genotypic diversity (G _o =17.0387) were very similar to Hardy–Weinberg expected frequencies (H _e =0.446; G ^* _e =17.0010). Evidence suggests that the contribution of asexual reproduction to recruitment in the study population of A. inornata is low. For a single population of K. commensalis from 4 850 m on the Porcupine Abyssal Plain, North Atlantic Ocean, the hypothesis that inbreeding due to reduced occurrence of outcrossing between anemones on a single holothurian host was examined by electrophoresis of ten randomly selected enzyme loci. Single-locus genotypic frequencies were significantly different from expected frequencies for one locus P≤ 0.05, hexokinase-1 (Hex-1)]. F _IS values were significantly different from zero for two enzyme loci (Hex-1 and Hex-2, P≤ 0.01 and P≤ 0.05, respectively), and the overall observed heterozygosity was lower than the expected heterozygosity (H _o =0.125, H _e =0.140). The hypothesis of inbreeding could not be rejected by the present study, although sample size was small (N=55), leading to possible bias in tests of significance. Genetic variation in A. inornata was higher than that recorded for most eukaryotes, although interlocus sampling error for only five loci is high. High genetic variability has been found in other sea anemones, and has been related to high longevity and mixed reproductive modes. Genetic variation in K. commensalis was in the higher range of that found in other eukaryotes, and is not unusual for anemones. Received: 5 August 1996 / Accepted: 11 December 1996     

Latitudinal variation in the Dungeness crab,Cancer magister: zoeal morphology explained by incubation temperature

The difference in morphology between zoeae of Cancer magister Dana from Alaskan and Californian waters was documented to determine if the morphological variation is attributable to environmental influences. First-stage zoeae from Alaska have significantly longer carapace spines than zoeae from central California. The dorsal, rostral and lateral carapace spines were 14, 14 and 29% longer, respectively, in the Alaskan zoeae. The effect of temperature was tested on zoeal morphology as it is an obvious environmental difference between Alaskan and Californian waters. Ovigerous female crabs collected in southeastern Alaska in 1984 were held at 1°, 5°, 10° and 15° C until hatching occurred. Eggs were sampled seven times during the incubation period, and relative mortality, egg diameter and development stage were measured. All of the crabs and eggs at 1° C died before hatching occurred. Egg mortality averaged less than 2% in the other temperature treatments. Egg diameter increased significantly over the incubation period for all temperatures. Developmental rate of the embryos was inversely related to temperature. Hatching first occurred in 42 d at 15° C, 60 at 10° C and 160 d at 5° C. Newly hatched zoeae were collected and body length, dorsal, rostral and lateral carapace spines were measured. Significant differences existed between all temperatures for all spine lengths, with longer spines occurring at lower temperatures. Zoeal body lengths were also significantly different between the three temperatures. The results of this study question the use of spine lengths to distinguish similar larval species.     

Assessment of the phototoxicity of weathered Alaska North Slope crude oil to juvenile pink salmon

Petroleum products are known to have greater toxicity to the translucent embryos and larvae of aquatic organisms in the presence of ultraviolet radiation (UV) compared to toxicity determined in tests performed under standard laboratory lighting with minimal UV. This study assessed the acute phototoxicity of the water accommodated fractions of weathered Alaska North Slope crude oil (ANS) to juvenile pink salmon, which are a heavily pigmented life stage. Fish in the highest ANS treatments exhibited melanosis, less mobility, reduced startle response, erratic swimming, and loss of equilibrium. Gills from fish exposed to ANS had elevated levels of hydroperoxides in oil-only, UV-only, and oil+UV treatments compared to control fish, which was indicative of increased lipid peroxidation in gill tissue. Under the test conditions of moderate salinity, low UV and high short-term oil exposure there were no indications of photoenhanced toxicity as assessed by elevation of mortality, behavioral impairment, or gill lipid peroxidation in oil+UV treatments. The results of this study suggest that pink salmon may be at less risk from photoenhanced toxicity compared to the translucent early-life stages of several other Alaska species.     

Net productivity of two cohorts of Chordaria flagelliformis (Phaeophyta) in Nova Scotia,Canada

Productivities of two cohorts of Chordaria flagelliformis (O. F. Müll.) C. Ag. were estimated from measured changes in biomass and survivorship over time. Maximum productivity during the summer growing season was 2.6 g C m^-2 d^-1. Although this figure is relatively high, the short growing season results in an annual production of only 89 g C m^-2. The significance of primary production by C. flagelliformis lies in its seasonal timing. During the summer growth period, 50% of production was recycled directly by detrital material. During the same time period, productivity and biomass losses of other seaweeds are at their lowest.     

Real-time PCR with molecular beacons provides a highly accurate assay for detection of Tay-Sachs alleles in single cells

The results presented here provide the first single-cell genetic assay for Tay-Sachs disease based on real-time PCR. Individual lymphoblasts were lysed with an optimized lysis buffer and assayed using one pair of primers that amplifies both the wild type and 1278 + TATC Tay-Sachs alleles. The resulting amplicons were detected in real time with two molecular beacons each with a different colored fluorochrome. The kinetics of amplicon accumulation generate objective criteria by which to evaluate the validity of each reaction. The assay had an overall utility of 95%, based on the detection of at least one signal in 235 of the 248 attempted tests and an efficiency of 97%, as 7 of the 235 samples were excluded from further analysis for objective quantitative reasons. The accuracy of the assay was 99.1%, because 228 of 230 samples gave signals consistent with the genotype of the cells. Only two of the 135 heterozygous samples were allele drop-outs, a rate far lower than previously reported for single-cell Tay-Sachs assays using conventional methods of PCR. Copyright © 2002 John Wiley & Sons, Ltd.     

The occurrence of glyphosate, atrazine, and other pesticides in vernal pools and adjacent streams in Washington, DC, Maryland, Iowa, and Wyoming, 2005–2006

Vernal pools are sensitive environments that provide critical habitat for many species, including amphibians. These small water bodies are not always protected by pesticide label requirements for no-spray buffer zones, and the occurrence of pesticides in them is poorly documented. In this study, we investigated the occurrence of glyphosate, its primary degradation product aminomethylphosphonic acid, and additional pesticides in vernal pools and adjacent flowing waters. Most sampling sites were chosen to be in areas where glyphosate was being used either in production agriculture or for nonindigenous plant control. The four site locations were in otherwise protected areas (e.g., in a National Park). When possible, water samples were collected both before and after glyphosate application in 2005 and 2006. Twenty-eight pesticides or pesticide degradation products were detected in the study, and as many as 11 were identified in individual samples. Atrazine was detected most frequently and concentrations exceeded the freshwater aquatic life standard of 1.8 micrograms per liter (microg/l) in samples from Rands Ditch and Browns Ditch in DeSoto National Wildlife Refuge. Glyphosate was measured at the highest concentration (328 microg/l) in a sample from Riley Spring Pond in Rock Creek National Park. This concentration exceeded the freshwater aquatic life standard for glyphosate of 65 microg/l. Aminomethylphosphonic acid, triclopyr, and nicosulfuron also were detected at concentrations greater than 3.0 microg/l.     

Role of Riparian Areas in Atmospheric Pesticide Deposition and Its Potential Effect on Water Quality

Riparian buffers are known to mitigate hydrologic losses of nutrients and other contaminants as they exit agricultural fields. The vegetation of riparian buffers can also trap atmospheric contaminants, and these pollutants can subsequently be delivered via rain to the riparian buffer floor. These processes, however, are poorly understood especially for pesticide residues. Therefore, we conducted a four‐year study examining stemflow and throughfall to a riparian buffer which was adjacent a cultured Zea mays field treated with atrazine and metolachlor. Stemflow is rain contacting the tree canopy traveling down smaller to larger branches and down the tree trunk, whereas throughfall is rain that may or may not contact leaves and branches and reaches the earth. Stemflow concentrations of the herbicides were larger than throughfall concentrations and accounted for 5‐15% of the atrazine and 6‐66% of the metolachlor depositional fluxes under the canopy. Larger depositional fluxes were measured when leaves were more fully emerged and temperatures and humidity were elevated. Rain collected outside the riparian buffer on the field side and on the back side revealed the trees trapped the herbicide residues. Herbicide loading to the riparian buffer stream was found to be linked to tree canopy deposition and subsequent washoff during rain events. These results indicate that in agricultural areas canopy washoff can be an important source of pesticides to surface waters.     

Fate of Triclosan and Methyltriclosan in soil from biosolids application

This study investigates the persistence of Triclosan (TCS), and its degradation product, Methyltriclosan (MeTCS), after land application of biosolids to an experimental agricultural plot under both till and no till. Surface soil samples (n = 40) were collected several times over a three years period and sieved to remove biosolids. Concentration of TCS in the soil gradually increased with maximum levels of 63.7 ± 14.1 ng g(-1) dry wt., far below the predicted maximum concentration of 307.5 ng g(-1) dry wt. TCS disappearance corresponded with MeTCS appearance, suggesting in situ formation. Our results suggest that soil incorporation and degradation processes are taking place simultaneously and that TCS background levels are achieved within two years. TCS half-life (t(0.5)) was determined as 104 d and MeTCS t(0.5), which was more persistent than TCS, was estimated at 443 d.