The role of GC-MS and LC-MS in the discovery of drinking water disinfection by-products

Gas chromatography-mass spectrometry (GC-MS) has played a pivotal role in the discovery of disinfection by-products (DBPs) in drinking water. DBPs are formed when disinfectants, such as chlorine, ozone, chlorine dioxide or chloramine, react with natural organic matter in the water. The first DBP known--chloroform--was identified by Rook in 1974 using GC-MS. Soon thereafter, chloroform and other trihalomethanes were found to be ubiquitous in chlorinated drinking water. In 1976, the National Cancer Institute published results linking chloroform to cancer in laboratory animals, and an important public health issue was born. Mass spectrometry and, specifically, GC-MS became the key tool used for measuring these DBPs in water and for discovering other DBPs that were formed. Over the last 25 years, hundreds of DBPs have been identified, mostly through the use of GC-MS, which has spawned additional health effects studies and regulations. Early on, GC with low resolution electron ionization (EI)-MS was used, together with confirmation with chemical standards, for identification work. Later, researchers utilized chemical ionization (CI)-MS to provide molecular weight information and high resolution El-MS to aid in the determination of empirical formulae for the molecular ions and fragments. More recently, liquid chromatography-mass spectrometry (LC-MS) with either electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) has been used to try to uncover highly polar DBPs that most experts believe have been missed by earlier GC-MS studies. Despite 25 years of research in the identification of new DBPs, new ones are being discovered every year, even for chlorine which has been the most extensively studied.     

Rain forest fragmentation and the proliferation of successional trees

The effects of habitat fragmentation on diverse tropical tree communities are poorly understood. Over a 20-year period we monitored the density of 52 tree species in nine predominantly successional genera (Annona, Bellucia, Cecropia, Croton, Goupia, Jacaranda, Miconia, Pourouma, Vismia) in fragmented and continuous Amazonian forests. We also evaluated the relative importance of soil, topographic, forest dynamic, and landscape variables in explaining the abundance and species composition of successional trees. Data were collected within 66 permanent 1-ha plots within a large (approximately 1000 km2) experimental landscape, with forest fragments ranging from 1 to 100 ha in area. Prior to forest fragmentation, successional trees were uncommon, typically comprising 2-3% of all trees (> or =10 cm diameter at breast height [1.3 m above the ground surface]) in each plot. Following fragmentation, the density and basal area of successional trees increased rapidly. By 13-17 years after fragmentation, successional trees had tripled in abundance in fragment and edge plots and constituted more than a quarter of all trees in some plots. Fragment age had strong, positive effects on the density and basal area of successional trees, with no indication of a plateau in these variables, suggesting that successional species could become even more abundant in fragments over time. Nonetheless, the 52 species differed greatly in their responses to fragmentation and forest edges. Some disturbance-favoring pioneers (e.g., Cecropia sciadophylla, Vismia guianensis, V. amazonica, V. bemerguii, Miconia cf. crassinervia) increased by >1000% in density on edge plots, whereas over a third (19 of 52) of all species remained constant or declined in numbers. Species responses to fragmentation were effectively predicted by their median growth rate in nearby intact forest, suggesting that faster-growing species have a strong advantage in forest fragments. An ordination analysis revealed three main gradients in successional-species composition across our study area. Species gradients were most strongly influenced by the standlevel rate of tree mortality on each plot and by the number of nearby forest edges. Species-composition also varied significantly among different cattle ranches, which differed in their surrounding matrices and disturbance histories. These same variables were also the best predictors of total successional-tree abundance and species richness. Successional-tree assemblages in fragment interior plots (>150 m from edge), which are subjected to fragment area effects but not edge effects, did not differ significantly from those in intact forest, indicating that area effects per se had little influence on successional trees. Soils and topography also had little discernable effect on these species. Collectively, our results indicate that successional-tree species proliferate rapidly in fragmented Amazonian forests, largely as a result of chronically elevated tree mortality near forest edges and possibly an increased seed rain from successional plants growing in nearby degraded habitats. The proliferation of fast-growing successional trees and correlated decline of old-growth trees will have important effects on species composition, forest dynamics, carbon storage, and nutrient cycling in fragmented forests.     

A Study of Wood Stove Particulate Emissions

Particulate emission factors for two wood stove models have been determined for two types of fuel and a range of operating conditions. The emission factors range from 1 g/kg (fuel) to 24 g/kg. A model is presented which represents the emission factor as a simple function of the ratio of fuel load to combustion rate, or the length of time between refueling. This model is felt to be appropriate for evaluating the impact of wood-based residential space heating on ambient air concentrations of particulate matter If certain assumptions can be made about stove operating conditions. An application of the emission factor model to a typical community suggests that the contribution of wood stoves to ambient particulate levels might reach 100 μg/m³ if the entire heating load were carried by wood.

Preliminary analyses of the particulate matter Indicate that benzene extractables range from 42% of the total particulate mass at short refuel times to 67% at longer refuel times. About 45% of the mass of benzene extractables appeared in the neutral fraction of acid base extractions. Polycyclic aromatic hydrocarbons are expected to be included in this neutral fraction.     

Methods to estimate solar radiation dosimetry in coral reefs using remote sensed, modeled, and in situ data

Solar irradiance has been increasingly recognized as an important determinant of bleaching in coral reefs, but measurements of solar radiation exposure within coral reefs have been relatively limited. Solar radiation dosimetry within multiple coral reef areas of South Florida was assessed using remote sensed, modeled, and measured values during a minor bleaching event during August 2005. Coral reefs in the Dry Tortugas and Upper Keys had similar diffuse downwelling attenuation coefficients (Kd, m(-1)), whereas Kd values were significantly greater in the Middle and Lower Keys. Mean 1% attenuation depths varied by reef region for ultraviolet B (UVB; 9.7 to 20 m), ultraviolet A (UVA; 22 to 40 m) and visible (27 to 43 m) solar radiation. Solar irradiances determined from remote sensed data were significantly correlated with measured values, but were generally overestimated at the depth of corals. Solar irradiances modeled using an atmospheric radiative transfer model parameterized with site specific approximations of cloud cover showed close agreement with measured values. Estimated daily doses (W h/m(2)) of UVB (0.01-19), UVA (2-360) and visible (29-1,653) solar radiation varied with coral depth (2 to 24 m) and meteorological conditions. These results indicate large variation in solar radiation dosimetry within coral reefs that may be estimated with reasonable accuracy using regional Kd measurements and radiative transfer modeling.     

On the use of log-transformation vs. nonlinear regression for analyzing biological power laws

Power-law relationships are among the most well-studied functional relationships in biology. Recently the common practice of fitting power laws using linear regression (LR) on log-transformed data has been criticized, calling into question the conclusions of hundreds of studies. It has been suggested that nonlinear regression (NLR) is preferable, but no rigorous comparison of these two methods has been conducted. Using Monte Carlo simulations, we demonstrate that the error distribution determines which method performs better, with NLR better characterizing data with additive, homoscedastic, normal error and LR better characterizing data with multiplicative, heteroscedastic, lognormal error. Analysis of 471 biological power laws shows that both forms of error occur in nature. While previous analyses based on log-transformation appear to be generally valid, future analyses should choose methods based on a combination of biological plausibility and analysis of the error distribution. We provide detailed guidelines and associated computer code for doing so, including a model averaging approach for cases where the error structure is uncertain.     

A novel statistical method for classifying habitat generalists and specialists

We develop a novel statistical approach for classifying generalists and specialists in two distinct habitats. Using a multinomial model based on estimated species relative abundance in two habitats, our method minimizes bias due to differences in sampling intensities between two habitat types as well as bias due to insufficient sampling within each habitat. The method permits a robust statistical classification of habitat specialists and generalists, without excluding rare species a priori. Based on a user-defined specialization threshold, the model classifies species into one of four groups: (1) generalist; (2) habitat A specialist; (3) habitat B specialist; and (4) too rare to classify with confidence. We illustrate our multinomial classification method using two contrasting data sets: (1) bird abundance in woodland and heath habitats in southeastern Australia and (2) tree abundance in second-growth (SG) and old-growth (OG) rain forests in the Caribbean lowlands of northeastern Costa Rica. We evaluate the multinomial model in detail for the tree data set. Our results for birds were highly concordant with a previous nonstatistical classification, but our method classified a higher fraction (57.7%) of bird species with statistical confidence. Based on a conservative specialization threshold and adjustment for multiple comparisons, 64.4% of tree species in the full sample were too rare to classify with confidence. Among the species classified, OG specialists constituted the largest class (40.6%), followed by generalist tree species (36.7%) and SG specialists (22.7%). The multinomial model was more sensitive than indicator value analysis or abundance-based phi coefficient indices in detecting habitat specialists and also detects generalists statistically. Classification of specialists and generalists based on rarefied subsamples was highly consistent with classification based on the full sample, even for sampling percentages as low as 20%. Major advantages of the new method are (1) its ability to distinguish habitat generalists (species with no significant habitat affinity) from species that are simply too rare to classify and (2) applicability to a single representative sample or a single pooled set of representative samples from each of two habitat types. The method as currently developed can be applied to no more than two habitats at a time.     

A test of the mismatch hypothesis: How is timing of reproduction related to food abundance in an aerial insectivore?

In seasonal environments, vertebrates are generally thought to time their reproduction so offspring are raised during the peak of food abundance. The mismatch hypothesis predicts that reproductive success is maximized when animals synchronize their reproduction with the food supply. Understanding the mechanisms influencing the timing of reproduction has taken on new urgency as climate change is altering environmental conditions during reproduction, and there is concern that species will not be able to synchronize their reproduction with changing food supplies. Using data from five sites over 24 years (37 site-years), we tested the assumptions of the mismatch hypothesis in the Tree Swallow (Tachycineta bicolor), a widespread aerial insectivore, whose timing of egg-laying has shifted earlier by nine days since the 1950s. Contrary to the mismatch hypothesis, the start of egg-laying was strongly related to food abundance (flying insect biomass) during the laying period and not to timing of the seasonal peak in food supply. In fact, food abundance generally continued to increase throughout the breeding season, and there was no evidence of selection based on the mistiming of laying with the seasonal peak of food abundance. In contrast, there was selection for laying earlier, because birds that lay earlier generally have larger clutches and fledge more young. Overall, initial reproductive decisions in this insectivore appear to be based on the food supply during egg formation and not the nestling period. Thus, the mismatch hypothesis may not apply in environments with relatively constant or abundant food throughout the breeding season. Although climate change is often associated with earlier reproduction, our results caution that it is not necessarily driven by selection for synchronized reproduction.     

Morphology and distribution of the ampullary electroreceptors in wobbegong sharks: implications for feeding behaviour

The electrosensory capabilities of wobbegong sharks are of particular interest, partly because very little is known about their behavioural ecology and specifically because of their unusual ambush predatory strategy and benthic lifestyle. While several biological functions of electroreception have been proposed, less consideration has been given to the functional significance of interspecific differences in the morphology and topographic distribution of the ampullary organs. The morphology of the ampullary organs was examined in four species of wobbegong shark, and the distribution of electroreceptive pores was mapped in two species. The ampullary systems of wobbegongs are similar in morphology to other marine elasmobranchs. The number of alveoli per ampullae is not significantly different between the four species; however, differences are seen between ampullary cell size in some species. Ampullary pore distribution patterns are relatively unique, with the majority of pores occurring on the dorsal region of the head. Wobbegongs feed primarily on demersal teleost fishes, and as the benthic and well-camouflaged wobbegong remains motionless, these fish could be easily detected by the dorsal pores when swimming within range.     

The influence of suburban land use on habitat and biotic integrity of coastal Rhode Island streams

Watershed land use in suburban areas can affect stream biota through degradation of instream habitat, water quality, and riparian vegetation. By monitoring stream biotic communities in various geographic regions, we can better understand and conserve our watershed ecosystems. The objective of this study was to examine the relationship between watershed land use and the integrity of benthic invertebrate communities in eight streams that were assessed over a 3-year period (2001-2003). Sites were selected from coastal Rhode Island watersheds along a residential land-use gradient (4-59%). Using the rapid bioassessment protocol, we collected biological, physicochemical, habitat, and nutrient data from wadeable stream reaches and compared metrics of structure and integrity. Principal component analyses showed significant negative correlation of indicators for stream physicochemical, habitat, and instream biodiversity with increasing residential land use (RLU) in the watershed. The physicochemical variables that were most responsive to percent RLU were conductivity, instream habitat, nitrate, and dissolved inorganic nitrogen (DIN). The positive correlation of DIN with percent RLU indicated an anthropogenic source of pollution affecting the streams. The biotic composition of the streams shifted from sensitive to insensitive taxa as percent RLU increased; the most responsive biological variables were percent Ephemeroptera, percent Scrapers, percent Insects, and the Hilsenhoff biotic index. These data show the importance of land management and conservation at the watershed scale to sustaining the biotic integrity of coastal stream ecosystems.     

Liver lesions in demersal fishes near a large ocean outfall on the San Pedro Shelf, California

The prevalence of toxicopathic liver lesions in demersal fish on the San Pedro Shelf, California was determined for a 15-year period (1988–2003). Fish livers were sampled at fixed locations as part of the Orange County Sanitation Districts (OCSD) ocean monitoring program. Histopathological examination of selected fish liver tissues was studied to determine whether the wastewater discharge had affected fish health. The prevalence of toxicopathic lesion classes neoplasms (NEO), preneoplastic foci of cellular alteration (FCA), and hydropic vacuolation (HYDVAC) varied among species and locations. For all species sampled, severe lesions occurred in 6.2% of the fish examined (n = 7,694). HYDVAC (4.1%) was the most common toxicopathic lesion type followed by FCA (1.4%) and NEO (0.7%). HYDVAC occurred only in white croaker (Genyonemus lineatus), accounting for 84.8% of the toxicopathic lesions for this species. Prevalence of HYDVAC, NEO, and FCA in white croaker was 15.2, 2.0, and 0.7%, respectively. The prevalence of HYDVAC and NEO in white croaker increased with age and size but there was no sexual difference. A linear regression model was used for hypothesis testing to account for significant differences in fish size (and age for croakers) at the different sampling locations. This analysis showed that for HYDVAC there was no spatial or location effect for lesion rate or size/age of onset. For NEO, the model predicted that white croaker near the wastewater outfall may acquire these lesions at a smaller size/younger age, and at a higher rate, than at other sites. However, this result may be biased due to the unequal size frequency distributions and the low prevalence of NEO in white croaker at the different sampling sites. Bigmouth sole (Hippoglossina stomata) had a prevalence of FCA and NEO of 1.3 and 0.35%, respectively, but the prevalence and distribution of lesions was too few for statistical testing. There was no sexual difference for lesion prevalence in hornyhead turbot (Pleuronichthys verticalis) and the prevalence of FCA and NEO was 3.4 and 0.37%, respectively. FCA prevalence increased with size in hornyhead turbot and there were no significant spatial differences for lesion rates and fish size at lesion onset. Overall, consistent spatial differences for lesion prevalence were not demonstrated and highlight the analytical difficulties of detecting a possible point source impact when the effect is rare, correlated with the size/age structure of the population, and may be caused by exposure to unknown multiple sources. Thus, the usefulness of liver histopathology as a point-source monitoring tool is best applied to where the spatial scale of impact generally exceeds the home range of the target species.