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The common bully (Gobiomorphus cotidianus) was used as a bioindicator to assess the impacts of geothermal, bleached kraft mill effluent (BKME), sewage, and thermal discharges in the Waikato River, Northern New Zealand. A range of morphological and physiological parameters were measured to assess fitness in fish sampled upstream and downstream from these point sources. Isotopic signatures gave evidence that fish were resident at sites and showed a gradient corresponding to site-specific sources of carbon and nitrogen along the river. There were changes in the size range and age structure of the populations but, no physiological responses were detected. Limited impacts were observed on common bully health at individual discharge sites, or downstream where the river showed gradual deterioration in water quality, particularly nutrient enrichment, and increased turbidity. Despite the gradual deterioration in water quality no concomitant cumulative impacts were observed in common bully. Responses were largely local in nature, responding to point-source discharges. Population responses at the geothermal and BKME discharge sites warrant further study to investigate possible effects on fish recruitment.

Brook trout populations in Prince Edward Island, Canada, have experienced over 50 pesticide-related fish kills since the 1960s. Life history evaluation of large sea-run brook trout recovered following two fish kill events was compared with a reference river using strontium:calcium otolith microchemistry. This study examined the dual hypotheses that anadromous brook trout are more likely to arise from sea-run mothers, and that freshwater entry timing makes them vulnerable to pesticide-induced fish kills. A total 89% of the fish exhibited an anadromous life history, and 77% of these were offspring of anadromous mothers, suggesting that anadromy is dominant in progeny of sea-run mothers. This study adds to our understanding of the maternal inheritance of anadromy in sea-run brook trout populations. Additionally, freshwater entry precedes the majority of fish kill events, illustrating that the overlap between migration and pesticide runoff contributes to the cumulative population risks to sea-run brook trout.

Estuaries are productive systems with rapid changes in natural stressors, such as salinity, that make perturbation detection challenging. Amphipods are vital to estuarine assemblages and may serve as anthropogenic stress indicators. However, practical methods of monitoring species composition and abundance are lacking. This study aims to describe the spatial patterns of four Gammarus spp. in northwest Atlantic estuaries and to compare their compositions and abundances between collection methods using artificial substrates, macrophyte raking, light-baited traps, and species-specific quantitative PCR analyses from sediment environmental DNA (eDNA). Sampling occurred in upper, mid, and lower estuary zones within three estuaries of Prince Edward Island (Canada). G. tigrinus was rarely found in the upper zones with any method. G. mucronatus was predominant in the upper–mid zones across the physical methods, and its abundance declined with increasing salinity. G. lawrencianus was a dominant species across zones, but its abundance did not change with salinity. G. oceanicus was predominant in lower-zone artificial substrates. Species abundances generally correlated with physical collection methods. Sediment eDNA did not detect the spatial effects observed via the physical methods but was correlated with the mean counts of G. mucronatus. The Gammarus spp. assemblages are spatially partitioned in short estuaries, though the sampling method is critical when interpreting estuary patterns. Though multiple methods are ideal for compositional comparisons, abundance monitoring should employ light traps.

Movement of anadromous fishes from marine to freshwater habitats provides a seasonal pulse of marine-derived nutrients (MDNs) to freshwater biota. In northeastern North America, iteroparous...

In this study, we examined the effects of dissolved oxygen, via metrics based on hourly measurements, and other environmental variables on invertebrate assemblages in estuaries spanning a gradient of nutrient loading and geography in the southern Gulf of St. Lawrence, Canada. Upper areas (15–25 practical salinity units (PSU)) of 13 estuaries that were dominated by either seagrass ( Zostera marina Linnaeus, 1753) or macroalgae ( Ulva spp. Linnaeus, 1753) were sampled from June to September 2013. Macroinvertebrate assemblages from Z. marina were found to be distinct from Ulva assemblages for both epifauna and infauna. Small snails dominated each vegetation type, specifically cerithids in Z. marina and hydrobids in Ulva. Although Z. marina had higher species richness, approximately 70% of species were common to both habitats. Faunal communities differed among estuaries with large, within-estuary, temporal variance only observed at Ulva sites impacted by hypoxia and particularly at sites with long water residence time. Indeed, abundances varied by several orders of magnitude in Ulva ranging from zero to thousands of macroinvertebrates. There was a strong negative correlation between hypoxic or anoxic water, 48 h prior to sampling, with relative abundances of amphipods, and a positive correlation with the relative abundances of snails. As one of the first studies to use high-frequency oxygen monitoring, this study revealed probable impacts and the transient nature of hypoxia in eutrophication.

Seagrasses are declining globally; understanding and monitoring these trends forms an important basis for management practices. This study examined eelgrass (Zostera marina) structural and isotopic indicators as a means of characterizing stress from the impacts of agricultural land use in four estuaries in the Southern Gulf of St. Lawrence, Canada. These estuaries ranged from 5 to 62% agricultural land use in their respective watersheds. Anthropogenic stressors such as increased nutrient loading and decreased light penetration (from sedimentation and phytoplankton) and natural stressors such as low salinity and temperature were measured concurrently with plant structural indicators, e.g., aboveground biomass, belowground biomass, canopy height, and root-to-shoot ratio, across the spatial range of eelgrass presence in each estuary. Plant isotopic indicators included leaf tissue concentrations of C, N, S, and stable isotopes thereof and were measured at each station in eelgrass leaf tissue. Eelgrass plant indicators varied significantly across the gradients within each estuary as well as throughout the growing season, but due to the very high within-estuary variability indicators did not vary significantly between estuaries. Eelgrass leaf nutrients and stable isotopes also varied significantly between stations within estuaries though in this case, among-estuary differences were detected. A distance-based linear model was employed to examine the ability of estuarine characteristics and plant isotopic and elemental variables to predict the plant structural indicators. Results showed that factors related to both salinity and nitrogen loading influenced the biomass and structure of this plant but in different ways. Belowground biomass most correlated to salinity, and aboveground biomass most related to nitrogen loading with a strong relationship with light attenuation. Overall, the high level of within-estuary variability, and the strong influence of salinity, suggests that comparison of plant structural indicators between estuaries may have limited utility as a monitoring tool; eelgrass coverage throughout an estuary may better reflect the influence of land use at a regional scale.

Community-based monitoring programs (CBMPs) are a cost-effective option to collect the long-term data required to effectively monitor estuaries. Data quality concerns have caused some CBMP datasets, which could fill knowledge gaps for aquatic ecosystems, to go unused. The Community Aquatic Monitoring Program (CAMP) is a CBMP that has collected littoral nekton assemblage data from estuaries in the southern Gulf of St. Lawrence since 2003. Concerns with the CAMP sampling design (station placement and numbers) have prevented decision-makers from using the data to inform estuary health assessments. This study tested if CAMP’s sampling design that accommodates volunteer participation provides similar information as a scientific sampling approach. Six CAMP stations and six stations selected using a stratified random design were sampled at ten estuaries. A permutational-MANOVA revealed nekton assemblages were generally not significantly different between the two sampling designs. The current six CAMP stations are sufficient to detect the larger differences in species abundances that may indicate differences in estuary condition. The predicted increase in precision (2%) with twelve stations is not substantive enough to warrant an increased sampling effort. CAMP’s scientific utility is not limited by station selection bias or numbers. Furthermore, well-designed CBMPs can produce comparable data to scientific studies.

To investigate impacts of proposed oil sands aquatic reclamation techniques on benthic fish, white sucker (Catostomus commersonii Lacepede, 1803) were stocked in 2 experimental ponds-Demonstration Pond, containing aged fine tailings capped with fresh water, consistent with proposed end-pit lake designs, and South Bison Pond, containing aged unextracted oil sands material-to examine the effects of unmodified hydrocarbons. White sucker were stocked from a nearby reservoir at both sites in May 2010 and sampled 4 mo later to measure indicators of energy storage and utilization. Comparisons were then made with the source population and 2 reference lakes in the region. After exposure to aged tailings, white sucker had smaller testes and ovaries and reduced growth compared with the source population. Fish introduced to aged unextracted oil sands material showed an increase in growth over the same period. Limited available energy, endocrine disruption, and chronic stress likely contributed to the effects observed, corresponding to elevated concentrations of naphthenic acids, aromatic compounds in bile, and increased CYP1A activity. Because of the chemical and biological complexity of these systems, direct cause-effect relationships could not be identified; however, effects were associated with naphthenic acids, polycyclic aromatic hydrocarbons, ammonia, and high pH. Impacts on growth have not been previously observed in pelagic fishes examined in these systems, and may be related to differences in sediment interaction.

Eutrophic aquatic habitats are characterized by the proliferation of vegetation leading to a large standing biomass that upon decomposition may create hypoxic (low-oxygen) conditions. This is indeed the case in nutrient impacted estuaries of Prince Edward Island, Canada, where macroalgae, from the genusUlva,form submerged ephemeral mats. Hydrological forces and gases released from photosynthesis and decomposition lead to these mats occasionally floating to the water’s surface, henceforth termed floating mats. Here, we explore the hypothesis that floating mats are refugia during periods of sustained hypoxia/anoxia and examine how the invertebrate community responds to it. Floating mats were not always present, so in the first year (2013) sampling was attempted monthly and limited to when both floating and submerged mats occurred. In the subsequent year sampling was weekly, but at only one estuary due to logistical constraints from increased sampling frequency, and was not limited to when both mat types occurred. Water temperature, salinity, and pH were monitored bi-weekly with dissolved oxygen concentration measured hourly. The floating and submerged assemblages shared many of the same taxa but were statistically distinct communities; submerged mats tended to have a greater proportion of benthic animals and floating mats had more mobile invertebrates and insects. In 2014, sampling happened to occur in the weeks before the onset of anoxia, during 113 consecutive hours of sustained anoxia, and for four weeks after normoxic conditions returned. The invertebrate community on floating mats appeared to be unaffected by anoxia, indicating that these mats may be refugia during times of oxygen stress. Conversely, there was a dramatic decrease in animal abundances that remained depressed on submerged mats for two weeks. Cluster analysis revealed that the submerged mat communities from before the onset of anoxia and four weeks after anoxia were highly similar to each other, indicating recovery. When mobile animals were considered alone, there was an exponential relationship between the percentage of animals on floating mats, relative to the total number on both mat types, and hypoxia. The occupation of floating mats by invertebrates at all times, and their dominance there during hypoxia/anoxia, provides support for the hypothesis that floating mats are refugia.

Estuarine eutrophication as a result of agricultural land use, including the use of chemical fertilizers, is increasing worldwide. Prince Edward Island (PEI), Canada has very high agricultural intensity by international standards with approximately 44% of the land area under production, and some watersheds in excess of 75% agricultural land-use. The type of agriculture is also intensive with primarily row crops that have high chemical fertilizer and pesticide usage. In light of these stressors, the hypothesis of this study was that mummichog (Fundulus heteroclitus) population parameters would change with point and nonpoint source pollution, and that multivariate statistics could be used to draw associations with specific stressors. Fish were sampled on a monthly basis from May through August at 7 estuaries spanning a range of land use, nutrient, and contaminant loadings. A suite of environmental variables were simplified into 3 principal components: PC1 representing agricultural land use, N loading, and plant habitat, PC2 being dominated by sediment sand and silt distribution, and PC3 largely reflecting P loading and sediment organic matter. There were significant differences in abundance of both adult and young-of-the-year mummichog, and these changes associated most strongly with PC1, the largely N-driven agricultural influences. In contrast, somatic variables such as liver and gonad size did not show strong association with the environmental quality principal component scores. The sand and silt PC2 appeared to have the opposite association with the biological data, with siltier environments correlating to older, larger, less dense populations of mummichog. Although pesticide residues were detected in estuarine sediment, there was no clear relationship between these and watershed agricultural intensity or biochemical indicators. There was, however, a strong relationship between agricultural environmental variables (PC1) and in vitro steroid production that is suggestive of a potential chemical effect. Eutrophication appeared to be a primary stressor affecting mummichog populations, as nutrient enrichment was associated with changes in habitat variables and these in turn were associated with high mummichog density. Thus, mummichog population demographics appear to have use as an indicator of adverse or worsening conditions in estuaries. We concluded that, based on the subset of environmental factors evaluated, the nonpoint–source inputs of sediments and nutrients exerted the greatest influence on mummichog populations in PEI estuaries. Integr Environ Assess Manag 2013;9:496–507. © 2013 SETAC

Historic deforestation has deprived many river systems of their natural wood loadings. To study the effects of the loss of wood from waterways, a field trial was conducted in three small forested streams in New Zealand. The objectives were to (i) examine differences in fish assemblages among wooded pools (where wood provided cover), open pools and riffles and (ii) measure the effects of wood removal on channel morphology and fish assemblages. In the first part of the study, no significant differences were found in total fish density among the three habitats. However, total fish biomass was significantly higher in wooded pools (64% of total fish biomass) compared with open pools and riffles. Mean density and biomass of banded kokopu (Galaxias fasciatus) and mean biomass of longfin eel (Anguilla dieffenbachii) were highest in wooded pools, whereas the density and biomass of bluegill bully (Gobiomorphus hubbsi) and torrentfish (Cheimarrichthys fosteri) were highest in riffles. In the second part of the study, wood was removed from a 200-m section (treatment) in each stream, significantly reducing pool area and increasing the proportion of channel area and length in riffles. At the habitat scale, banded kokopu and large longfin eel were the two species mostly affected by wood removal. At the reach scale, banded kokopu biomass was significantly lower in the treatment sections. Although wooded pools were a small portion of total habitat, they provided important habitat for two of New Zealand's larger native fish taxa.

Artificial fertilizers are contributing to the replacement of eelgrass (Zostera marina) by sea lettuce (Ulva lactuca) in estuaries of Prince Edward Island (PEI), Canada. In this study, we found that the nearshore fish community differed between areas dominated by these two vegetations within an estuary in every month sampled (April–August). Adult northern pipefish (Syngnathus fuscus), threespine stickleback (Gasterosteus aculeatus), blackspotted stickleback (Gasterosteus wheatlandi), and Atlantic silverside (Menidia menidia) were most strongly associated with eelgrass, while mummichog (Fundulus heteroclitus), ninespine stickleback (Pungitius pungitius), and American eel (Anguilla rostrata) were often more numerous in sea lettuce. Sea lettuce stations tended to have more young-of-the-year mummichog, fourspine stickleback (Apeltes quadracus), and Gasterosteus sp. than eelgrass stations but fewer young-of-the-year northern pipefish and Atlantic silverside. Fish richness and abundance were significantly lower in the sea lettuce than eelgrass habitat during August when benthic hypoxia occurred. We conclude that the loss of eelgrass from PEI estuaries will result in significant declines in fish biodiversity.

To evaluate the effects of debris dams on aquatic invertebrate communities, we sampled benthic invertebrates in debris dams and riffles in three forested headwater streams in New Zealand. As part of a large-scale field experiment, debris dams were subsequently removed from three treatment sections in each of the streams to assess effects on invertebrate communities. Prior to debris dam removal, total invertebrate densities in debris dams were not significantly different from those in riffles. However, densities of Ephemeroptera, Plecoptera and Trichoptera taxa were significantly higher in debris dams than in riffles. Debris dams contained a higher number of less common taxa (defined as

The effluent of the Tasman pulp and paper mill (Kawerau, New Zealand) has been intensively studied for its effects on the health of fishes between 1998 and present. This review summarizes peer-reviewed scientific literature on the reproductive effects of the Tasman Mill effluent on fishes. In the 1990s there was an emerging body of literature from around the world showing that exposure to pulp and paper effluent could cause subtle reproductive alterations in exposed fishes. Locally, the Tarawera River had proved to be a difficult environment to conduct field studies. To overcome some of the difficulties with studying fish populations in the Tarawera River, initial studies on the reproductive health of fishes were focused on mesocosm and laboratory bioassays. During the later part of this period of study, wild fish population sampling was conducted inriver to assess the cumulative impact of multiple discharges. The initial mesocosm studies were conducted with rainbow trout exposures over an entire reproductive development cycle. The Tasman Mill effluent was initially observed to cause reductions in gonad size in females corresponding with lower circulating sex steroid hormones and reduced egg and larval sizes. This result was not observed again in the two subsequent long-term exposures conducted after 2001. Laboratory studies initially found the effluent to have a masculinizing effect on female mosquitofish (Gambusia affinis). This mosquitofish masculinization response disappeared after 2001 and was also not seen in effluent-exposed wild populations. Upstream and downstream populations of the native common bully (Gobiomorphus cotidianus) showed different reproductive timing, and investigation revealed that genetic differences were a potential reason for these differences. Subsequent investigation compared the Tarawera River bully to genetically similar Rangitiki River bully and found no evidence of reproductive alterations. The entire body of published data was assessed with regards to changes at the mill and chemical profiles of the effluent. It was evident that continuing effort on the part of the mill has resulted in gradual improvement in effluent quality over the duration of the studies. However, the disappearance of reproductive effects as assessed by multiple bioassays corresponds to one major change: screen room closure in the pulp mill. This change would have resulted in wood extractives being shunted from the treatment system to the recovery boiler, resulting in a net reduction in compounds derived from wood.

This study examined the comparative physiological health of the endemic New Zealand common bully (Gobiomorphus cotidianus) in the Tarawera and Rangitaiki Rivers. Bully were sampled downstream of pulp and paper effluent inputs in the Tarawera River and compared with a similar inland population in the Rangitaiki River. Condition factor and liver somatic index did not differ between populations, but Tarawera River bullies possessed larger gonads. Haematological assessments found smaller erythrocytes with reduced haemoglobin content, and increased leukocyte concentrations in Tarawera River females. Male and female Tarawera River bullies had significantly induced hepatic ethoxyresorufin-O-deethylase (EROD) activity. Greater ovarian follicular steroid production was also found for Tarawera River females. Microscopic analysis of ovarian tissue samples found no histopathological abnormalities in either population and indicated a slightly advanced vitellogenic stage of development in the Tarawera River population. It is concluded that the difference in steroid production between populations was most likely linked to gonad size and developmental status. In line with the disappearance of physiological effects in recent controlled laboratory and mesocosm pulp and paper effluent exposures, this study further demonstrates that, with the exception of EROD induction, characteristic pulp and paper effluent effects are not obvious in wild Tarawera fish.

The use of multiple spawning fishes in environmental effects monitoring programs has proven difficult for a number of reasons including the inability to predict reproductive investment and ensure synchronous sampling of reference and impacted populations. The estuarine resident northern mummichog (Fundulus heteroclitus macrolepidotus) has been successfully used as a sentinel for effects of pulp and paper mill effluents in Atlantic Canada and has been proposed for monitoring other anthropogenic impacts. This study investigated the spatial and temporal variability of the somatic parameters used to describe fish performance, specifically measures of energy use and storage, in estuaries located in Prince Edward Island, Canada. Three sites with varying levels of agricultural input were studied. Fish at all sites depleted their energy reserves over winter, as reflected in depressed condition, liver size, and gonad size, but then quickly replenished them in May. These population parameters were highly variable throughout the reproductive season and within an estuary. Spawning was continuous at all sites without indication of lunar or other periodicity. We conclude that repeated sampling is required to assess reproductive output in the northern mummichog and densities of adults and young-of-the-year deserve further investigation as a potentially less logistically demanding indicator of eutrophication.

The common bully (Gobiomorphus cotidianus), a smallbodied New Zealand native fish species, was used to monitor population impacts of multiple effluents in the Tarawera River, New Zealand. In an initial survey, the absence of reproductive development at the expected spawning time for common bully was observed in a population downstream of effluent discharges. Subsequently, we examined the hypotheses that the observed changes were due to effluent exposure, migratory patterns, or genetic differences between populations. Liver detoxification enzyme activity and stable isotopes provided evidence against upstream migration of sexually mature bully. The observed presence of developed gonads in the downstream population during winter season resulted in the rejection of the hypothesis that reproductive failure was due to effluent exposure, and it was concluded that there were substantial differences in reproductive timing. Genetic analyses of two upstream, one downstream, and one population from a nearby coastal river indicated the upstream (reference) and downstream (effluent exposed) bully in the river formed genetically distinct populations. The identification of a nearby river population with similar reproductive timing and high genetic similarity to the effluent-exposed population suggests that the observed differences in the genetics of the downstream population were not caused by effluent exposure. The genetic analysis did highlight the lack of donwstream dispersion and gene flow in the river which could possibly be related to anthropogenic stress.

The acute lethality of low dissolved oxygen (DO) was examined in laboratory studies using several New Zealand freshwater fish and two invertebrates at 15°C. The 48-h LC50 value was used as the endpoint for acute DO sensitivity as, owing to rapid mortality, this was found to best approxi- mate the threshold lethal concentration. Median le- thal time to death did not provide a reliable endpoint for comparing sensitivities. Fish LC50 values varied from 0.54 to 2.65 mg litre -1 , with inanga whitebait (Galaxias maculatus; 2.65 ± 0.19 mg litre -1 , mean ± SEM) being the most sensitive species tested. Common smelt (Retropinna retropinna; 1.83 ± 0.08 mg litre -1 ) and rainbow trout (Oncorhynchus mykiss; 1.61 ± 0.06 mg litre -1 ) were similar in their sensi- tivities, whereas common bully (Gobiomorphus cotidianus; 0.91 ± 0.06 mg litre -1 ) and shortfin eel elvers (Anguilla australis; 0.54 ± 0.03 mg litre -1 ) were the most tolerant fish. The shrimp (Paratya curvirostris; 0.82 ± 0.09 mg litre -1 ) and freshwater crayfish (Koura, Paranephrops planifrons; 0.77 ± 0.06 mg litre -1 ) were also tolerant to low DO. A subset of experiments to determine the relative sensitivities of larval and juvenile trout and bully in- dicated no significant differences between these life stages. Keywords dissolved oxygen; fish; freshwater; hypoxia; Anguilla australis; Gobiomorphus cotidianus; Retropinna retropinna; Paratya curvirostris; Paranephrops planifrons; Oncorhynchus mykiss; Galaxias maculatus

Rainbow trout were exposed to a secondary treated, thermomechanical/bleached kraft pulp and paper effluent in 12,000-L, flow-through exposure tanks at an environmental research facility located at a pulp and paper mill in Kawerau, New Zealand. Trout (age, 2+ years) were obtained from a local hatchery and exposed either to upstream river water or a nominal concentration of 12% (v/v) effluent diluted in upstream river water. Three treatment groups were used: Effluent exposure that started approximately three months before gonadal growth (eight-month total exposure), effluent exposure that started approximately halfway through gonadal development (two-month total exposure), and trout exposed to reference water alone for the total duration of the experiment. Trout were sacrificed just before spawning; exposure, growth, and reproductive endpoints were assessed during and at the termination of the experiment. Reduction in growth was observed in both sexes in the eight-month treatment group relative to the river water reference treatment group. No differences were observed in condition factor or liver size in either treatment. Females in the eight-month exposure group also had significantly lower ovary weight. The two-month exposure group showed no differences from the reference group in growth or somatic indices. Estradiol and testosterone were reduced in blood samples taken from the eight-month exposure group by four months into the experiment as compared to the reference treatment. Steroid and vitellogenin levels in individual female trout from this treatment were significantly correlated with gonadosomatic indices (GSI) measured at the termination of the experiment. The GSI was not correlated strongly or consistently with pregnenolone, nor were any treatment-related pregnenolone differences observed, indicating that the steroid hormone reductions likely were not related to cholesterol side-chain cleavage. Male trout showed significant induction of vitellogenin and lower I 1-ketotestosterone during the experiment (only the eight-month group was examined), but this did not result in any significant differences in testes development. Thus, this study has shown an impact of pulp mill effluent exposure on the reproductive physiology of female trout that appeared to be hormonally mediated. Furthermore, the effect could only be manifest when the exposure was initiated before the start of gonad development.

Northern leopard frogs (Rana pipiens) and green frogs (Rana clamitans) were evaluated at eight wetland sites, four of which were within apple orchards, to determine if environmental changes associated with orchard management affected measured biological parameters. Size, age, genetic variation, condition indices, levels of circulating steroid hormones, 7-ethoxyresorufin-O-deethylase activity (EROD), and organochlorine and organophosphorus residues in breeding males sampled at pond sites in orchards were compared to the same parameters measured in breeding males from reference sites. Also, the size and physiological condition of young-of-the-year captured in orchard and reference ponds were compared. No evidence of a reduction in genetic variation was found in populations of either species at any sites, but unexpectedly high average heterozygosity values (0.191–0.282) in concert with low overall fixation indices (0.012–0.059) in adults of both species did suggest that pond populations were interacting with neighboring populations in nonorchard habitats. Few significant differences in levels of circulating steroid hormones or condition indices of breeding males were found among sites. Significant EROD induction in male green frogs collected from one orchard site during one sampling event was the only indication that a metabolic challenge due to presence of cytochrome P450-inducing toxicants may have existed, whereas elevated concentrations of organochlorines (dichlorodiphenyltrichloroethane [DDT]- or endosulfan-related) in green frog tissues suggested that frogs at three orchard sites were taking up pesticides. Significant differences in size of equivalent-age male and juvenile leopard frogs and green frogs occupying different study sites suggested that suboptimal habitat characteristics existed at one or two of the four orchard sites. However, site-specific habitat deficiencies could not be related to orchard study sites in general, and, thus, wetlands in apple orchards appeared to provide viable breeding habitat for both northern leopard frogs and green frogs.

Over a 4-year period from 1990 to 1993, the H4IIE bioassay was used to measure 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalent concentrations (TECs) in the liver tissue of white sucker exposed to bleached kraft mill effluent. During two of the sampling periods, the H4IIE bioassay-derived TECs (BD-TECs) were compared directly with BD-TECs measured by a similar bioassay using a rainbow trout liver cell line (RTL-W1). From 1989 to 1993, chemistry-derived TECs (CD-TECs) were calculated directly from measured polychlorinated dibenzo-p-dioxin (PCDD) and dibenzofuran (PCDF) concentrations in liver tissue. Chemistry-derived TECs were compared to bioassay-derived TECs using toxic equivalent factors (TEFs) developed for these systems. During the study period, the pulp mill began secondary treatment of effluent and increased the substitution of chlorine dioxide for molecular chlorine during bleaching. Significant reductions in H4IIE BD-TECs and CD-TECs were observed in liver tissue of both male and female white sucker during summer and fall sampling periods but not during spring sampling periods. Process and treatment improvements at this particular bleached kraft pulp mill appear to have been successful at reducing the levels of PCDDs and PCDFs found in white sucker exposed to the effluent. Despite the decreases in TECs, relative mixed function oxygenase (MFO) induction in the exposed white sucker did not decrease over the study period, suggesting the PCDDs and PCDFs are currently not the dominant MFO-inducing compounds in the effluent of this mill.

While large numbers of hatchery reared salmonids have been stocked into the Great Lakes, few of these introduced populations have become self-substaining. Many salmonid populations are contaminated by organochlorine chemicals, and some experience embryonic and fry mortality when reared in hatcheries, which might reduce the survival of naturally deposited embryos and thereby compromise natural recruitment. Embryos from two such populations inhabiting Lake Ontario, coho salmon (Oncorhynchus kisutch) and chinook salmon (Oncorhynchus tshawytscha), were reared under hatchery conditions in 1990, experiencing mean mortality rates of 46% and 24% respectively. This mortality occurred both prior to eye-up and from eye-up to hatch, was female specific, and did not correlate with either muscle or egg levels of total PCB, mirex, octachlorostyrene, or DDT. The levels of 2,3,7,8-tetra-chlorodibenzo-p-dioxin equivalent concentrations (TEC) determined with the H41IE rat hepatoma bioassay ranged from 88 to 320 ppt in the eggs, and while they correlated with organochlorine residues, TEC did not correlate with embryonic mortality. Further investigations of fish-specific TEC bioassays and chemical or biotic influences on gonadal maturation and quality appear necessary to confirm the suggestion that embryonic mortality in these species is not related to organochlorines such as PCBs or dioxins.

Background Many postglacial lakes contain fish species with distinct ecomorphs. Similar evolutionary scenarios might be acting on evolutionarily young fish communities in lakes of remote islands. One process that drives diversification in island freshwater fish species is the colonization of depauperate freshwater environments by diadromous (migratory) taxa, which secondarily lose their migratory behaviour. The loss of migration limits dispersal and gene flow between distant populations, and, therefore, is expected to facilitate local morphological and genetic differentiation. To date, most studies have focused on interspecific relationships among migratory species and their non-migratory sister taxa. We hypothesize that the loss of migration facilitates intraspecific morphological, behavioural, and genetic differentiation between migratory and non-migratory populations of facultatively diadromous taxa, and, hence, incipient speciation of island freshwater fish species. Results Microchemical analyses of otolith isotopes (88Sr, 137Ba and 43Ca) differentiated migratory and non-migratory stocks of the New Zealand endemic Gobiomorphus cotidianus McDowall (Eleotridae). Samples were taken from two rivers, one lake and two geographically-separated outgroup locations. Meristic analyses of oculoscapular lateral line canals documented a gradual reduction of these structures in the non-migratory populations. Amplified fragment length polymorphism (AFLP) fingerprints revealed considerable genetic isolation between migratory and non-migratory populations. Temporal differences in reproductive timing (migratory = winter spawners, non-migratory = summer spawners; as inferred from gonadosomatic indices) provide a prezygotic reproductive isolation mechanism between the two ecotypes. Conclusion This study provides a holistic look at the role of diadromy in incipient speciation of island freshwater fish species. All four analytical approaches (otolith microchemistry, morphology, spawning timing, population genetics) yield congruent results, and provide clear and independent evidence for the existence of distinct migratory and non-migratory ecotypes within a river in a geographically confined range. The morphological changes within the non-migratory populations parallel interspecific patterns observed in all non-migratory New Zealand endemic Gobiomorphus species and other derived gobiid taxa, a pattern suggesting parallel evolution. This study indicates, for the first time, that distinct ecotypes of island freshwater fish species may be formed as a consequence of loss of migration and subsequent diversification. Therefore, if reproductive isolation persists, these processes may provide a mechanism to facilitate speciation.

The responses of shortfin eel (Anguilla australis) to discharges from two pulp and paper mills, municipal wastewater, and a geothermal power plant wastewater were examined. Eels were caged at 3 sites along the Tarawera River, North Island, New Zealand, to explore effects of a 3-wk exposure down a contamination gradient (Ref --D1 --D2). Most of the observed effects were seen in eels caged at the furthest downstream site (D2), below all the discharge areas. General hematology in eels was unaffected, as measures did not differ markedly at the two downstream sites compared with the reference site. At D2, eels were significantly lighter per unit length (reduced condition factor), although liver and spleen size (LSI and SSI) were unaffected. Significantly elevated circulating sex steroid concentrations (testosterone and estradiol) were measured in D2 eels and increasing sex steroid levels at both sites downstream of the reference site were observed. Significant ethoxyresorufin O-deethylase (EROD) activity induction was seen in D2 eels and bile chemistry showed significant accumulation of pyrene and retene equivalents. However, significantly greater concentrations of total resin acids were found in the bile of eels from the intermediate site (D1), between the two pulp and paper mills. The higher polycyclic aromatic hydrocarbon (PAH) equivalents found in the bile of D2 eels suggest that resin acid neutrals, particularly retene, are responsible for some of the effects observed in eels at the furthest downstream exposure site. Levels of pulp and paper mill extractives in sediment, including the PAH retene, support this conclusion.

The effects of point-source and diffuse discharges on resident populations of brown bullhead catfish (Ameiurus nebulosus (LeSueur, 1819)) in the Waikato River (New Zealand) were assessed at sites both upstream and downstream of point-source discharges. At each site, the population parameters, relative abundance, age structure, and individual indices, such as condition factor, organ (gonad, liver, and spleen) to somatic weight ratios, and number and size of follicles per female, were assessed. Physiological (blood), biochemical (hepatic ethoxyresorufin-O-deethylase [EROD] and plasma steroids), and other indicators (bile chemistry and liver metals) of exposure or response also were measured. No impacts on brown bullhead health were obvious at individual geothermal, municipal sewage, or thermal discharge sites or cumulatively along the river. Brown bullhead from the bleached kraft mill effluent site showed elevated levels of EROD, decreased numbers of red blood cells, increased numbers of white blood cells, and depressed levels of sex steroids. However, growth rates, condition factor, age structure, and gonadosomatic index suggest that discharges with significant heat or nutrients benefit catfish despite physiological impairment at one site. Consideration of brown bullhead population-level responses to discharges in a monitoring framework revealed three different population-level response patterns resulting from the point-source discharges.

Experiments were conducted to examine the effects of a thermomechanical (TMP)/bleached kraft pulp and paper mill effluent (BKME), dehydroabietic acid (DHAA), hypoxia, and combinations of hypoxia and effluent on juvenile rainbow trout. In the first two experiments, trout were exposed for 4 weeks to 0%, 10%, 30%, and 70% TMP/BKME or 0, 35, 110, and 250 microgL(-1) DHAA, respectively. Endpoints of those dose-response studies included critical swimming speed, oxygen consumption, and hematology. Reduced swimming performance was found for fish exposed to 70% TMP/BKME. Moderate increases in mean cell hemoglobin concentration at 70% TMP/BKME and blood glucose at 30% and 70% TMP/BKME were also seen. The opposite trend for glucose was found for DHAA-exposed fish, where a slight decrease in glucose was seen at 110 and 250 microgL(-1) DHAA. The third experiment examined the effects of 15% v/v TMP/BKME exposure at 2.5 and 5.0 mgL(-1) dissolved oxygen (DO) for 4 weeks. This experiment found no effect of low DO on swimming ability. An interactive effect between DO and effluent exposure was seen only on hematocrit, where effluent caused an increase in hematocrit at 5 mgL(-1) and a decrease at 2.5 mgL(-1) DO. Effluent exposure in this experiment resulted in a greater number of smaller red blood cells. The current study demonstrated physiological effects in rainbow trout exposed to varying concentrations (15-70% v/v) of a TMP/BKME and no substantial effects of DHAA exposure. With the exception of the reduced swimming performance in fish exposed to TMP/BKME, the observed effects are considered relatively small in magnitude but are occurring at concentrations of effluent that occur in the receiving environment.

The objective of the present study was to investigate the influence of chronic exposure to municipal sewage treatment effluent at environmentally relevant concentrations on immune parameters in rainbow trout (Oncorhynchus mykiss), including the assessment of potential differences in reactivity between sexually mature male and female fish. Trout were exposed to 1.5 and 15% (v/v) secondary treated municipal sewage effluent for 32 weeks. Fish were injected intra-peritoneally either with inactivated Aeromonas salmonicida to simulate an infection or with PBS as control for this immune challenge 6 weeks prior to sampling. Exposure to effluent resulted in a decrease in A. salmonicida-specific serum antibody level and blood lymphocyte numbers in mature females, but not in male fish. Injection of A. salmonicida resulted in enhanced serum lysozyme activity in mature male trout, which were not exposed to effluent. This stimulating effect of A. salmonicida could not be found in effluent-exposed trout, again potentially revealing a suppressive effect of the effluent. An influence of sampling fish on two consecutive days was observed in many immune parameters, most likely reflecting handling stress. Leucocyte and lymphocyte numbers in peripheral blood were consistently lower in male and female fish on the second sampling day. Phagocytosis in head kidney macrophages from male trout was also influenced by sampling day, whereby a stimulation of this reaction occurred on the second day of sampling. Liver mixed function oxygenase activity was found to be enhanced in mature male trout exposed to 15% effluent. In conclusion, the study showed, that exposure to sewage treatment plant effluent, in surface water relevant concentrations, can lead to potentially adverse effects on selected immune reactions in rainbow trout. However, this study also demonstrated that both handling stress and the sex of mature fish have distinct influences on the immune response detected in male and female fish and are likely to influence measured immune parameters to the extent that subtle effluent induced changes may be difficult to detect.

Biological Indicators of Aquatic Ecosystem Stress. Edited by S. M. Adams. American Fisheries Society, Bethesda, Maryland. 2002. 644 pages. $89.00 (cloth); $69.00 (paper) ($62.00 and 48.00 to AFS me...

For livers of lake trout from three Great Lakes sites, the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalent concentrations (TEQs) were determined in four ways. Bioassay-derived (BD) TEQs were measured with rat (H4IIE) and rainbow trout (RTL-W1) cell lines, and chemistry-derived (CD) TEQs were calculated with TCDD equivalency factors (TEFs) derived with H4IIE and RTL-W1. Generally, BD-TEQs and CD-TEQs for individual samples did not differ significantly for either H4IIE or RTL-W1, indicating that all of the AhR-active compounds in a sample were accounted for by the congener analysis and that these compounds acted in an additive fashion in both mammalian and piscine systems. However, contributions of individual chemicals to overall CD-TEQs differed with H4IIE and RTL-W1 TEFs, and for some individual samples, TEQs did depend on the method used. Hepatic TEQs and ethoxyresorufin-O-deethylase (EROD) levels differed significantly between sites. For TEQs, the order was Glenora > Jackfish Bay > Black Bay; for EROD activity, the order was Jackfish Bay > Glenora = Black Bay. No correlation was found between hepatic TEQs and EROD activity, which suggests that the two measurements are evaluating different but related consequences of contaminant exposure.

Levels of selected non-, mono-, and di-ortho-substituted polychlorinated biphenyl (PCB) congeners, polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) were determined in 4-year-old lake trout from the eastern basin of Lake Ontario, collected from 1977 to 1993. Results confirm that overall levels of contaminants have decreased steadily in lake trout since 1977, and that coplanar PCB levels do not appear to be increasing over time in relation to levels of other PCBs. Contaminant levels in lake trout from 3 to 9 years old, collected in 1988 from the western end of Lake Ontario, show the body burden of contaminants increases with age. Relative levels of coplanar PCBs to other PCBs for the age study samples showed no change, except for PCB 77, which exhibited a slight decrease in relation to total PCB levels. Toxic equivalents (TEQs) were calculated from fish contaminant concentrations measured for the time study using toxic equivalence factors (TEFs) from both mammalian and teleost studies. The relative contributions of PCBs, PCDDs, and PCDFs to total TEQs were examined. When TEFs used for risk assessment are applied to temporal trend data, 15 to 20% of the total TEQs were due to mono-ortho-substituted PCBs; 40 to 50% to non-ortho coplanar PCBs; and 20 to 30% to 2,3,7,8-substituted PCDD and PCDFs. The TEQs determined from lake trout extracts by an H4IIE cell bioassay technique are compared to those determined by chemical analyses, using a variety of TEFs.