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Wildlife ecological risk assessment in the 21st century: Promising technologies to assess toxicological effects

Despite advances in toxicity testing and development of new approach methodologies (NAMs) for hazard assessment, the ecological risk assessment (ERA) framework for terrestrial wildlife (i.e., air-breathing amphibians, reptiles, birds, and mammals) has remained unchanged for decades. While survival, growth, and reproductive endpoints derived from whole animal toxicity tests are central to hazard assessment, non-standard measures of biological effects at multiple levels of biological organization (e.g., molecular, cellular, tissue, organ, organism, population, community, ecosystem) have potential to enhance the relevance of prospective and retrospective wildlife ERAs. Other factors (e.g., indirect effects of contaminants on food supplies and infectious disease processes) are influenced by toxicants at individual, population, and community levels, and need to be factored into chemically-based risk assessments to enhance the “eco” component of ERAs. Regulatory and logistical challenges often relegate such non-standard endpoints and indirect effects to post-registration evaluations of pesticides and industrial chemicals, and contaminated site evaluations. While NAMs are being developed, to date their applications in ERAs focused on wildlife have been limited. No single magic tool or model will address all uncertainties in hazard assessment. Modernizing wildlife ERAs will likely entail combinations of laboratory and field-derived data at multiple levels of biological organization, knowledge collection solutions (e.g., systematic review, adverse outcome pathway frameworks), and inferential methods that facilitate integrations and risk estimations focused on species, populations, interspecific extrapolations, and ecosystem services modeling, with less dependence on whole animal data and simple hazard ratios.

Integrated Environmental Assessment and Management

Establishing appropriate measures for monitoring aging in birds: comparing short and long lived species

In order to reveal patterns of reproductive aging in birds we focus on a short lived species, the Japanese quail and the American kestrel, which has a life span of medium length. Quail have been studied extensively in the laboratory as models for understanding avian endocrinology and behavior, and as a subject for toxicological research and testing. In the lab, Japanese quail show age-related deterioration in endocrine, behavioral, and sensory system responses; the American kestrel is relatively long lived and shows moderate evidence of senescence in the oldest birds. Using data collected from captive kestrels at the Patuxent Wildlife Research Center, a database was designed to document selected parameters over the life cycle of the kestrels. Life table data collated from many species indicate that longer lived species of birds show senescence in survival ability but this pattern has not been established for reproductive function. We suggest that useful comparisons among species can be made by identifying stages in reproductive life history, organized on a relative time scale. Preliminary data from quail and kestrels, admittedly only two species, do not yet indicate a pattern of greater reproductive senescence in longer-lived birds.

Experimental Gerontology

The effects of salinity exposure on multiple life stages of a common freshwater mussel, Elliptio complanata

There is growing concern over the effects of increased salinization on freshwater organisms, which are largely unknown for unionid mussels. Adult and larval Elliptio complanata were exposed to low-level salt concentrations to determine the effects on mussel survival, physiology, and reproduction. Adults were exposed to salt concentrations of 0 parts per thousand (ppt), 2 ppt, 4 ppt, and 6 ppt NaCl and monitored over 7 d for mortality. Treatment groups exposed to 6 ppt and 4 ppt experienced 50% mortality at day 3 and day 4, respectively, with complete mortality by day 7. No mortality was observed in the other treatments. Adults were also exposed to sublethal salinity levels of 1 ppt and 2 ppt NaCl for 4 wk to determine physiological consequences of prolonged salinity exposure. Mussels exposed to 1 ppt and 2 ppt experienced reduced metabolic rates within the first 24 h of exposure that recovered to control levels in the 1-ppt treatment within 7 d. Metabolic recovery did not occur in the 2-ppt treatment by the end of 28 d. Glochidia exposed to 3-ppt NaCl during attachment to their host fish suffered a reduction in attachment success and metamorphosis, resulting in a 10-fold reduction in the number of juveniles produced per host fish. The present study demonstrates that low levels of salt can have a dramatic effect on the reproduction, physiology, and survival of freshwater mussels

Environmental Toxicology and Chemistry

Association of mercury and selenium with altered glutathione metabolism and oxidative stress in diving ducks from the San Francisco Bay region

Ault male greater scaup ( Aythya marila ), surf scoters ( Melanitta perspicillata ), and ruddy ducks ( Oxyurajamaicensis ) were collected from Suisun Bay and coastal Tomales Bay in the greater San Francisco Bay area to assess exposure to inorganic contaminants. Hepatic Se concentrations were highest in greater scaup (geometric mean = 67 ppm dry weight) and surf scoters (119 ppm) in Suisun Bay, whereas hepatic Hg was highest (19 ppm) in greater scaup and surf scoters from Tomales Bay. Hepatic Se and Hg were lower in ruddy ducks and did not differ between locations. Hepatic supernatants were assayed for enzymes related to glutathione metabolism and antioxidant activity, including glucose-6-phosphate dehydrogenase (G-6-PDH), glutathione peroxidase (GSH peroxidase), glutathione reductase (GSSG reductase), and glutathione- S -transferase (GSH transferase). Glutathione peroxidase activity was higher in surf scoters and ruddy ducks, and G-6-PDH was higher in greater scaup and surf scoters from Suisun Bay than Tomales Bay. Glutathione reductase (GSSG) was higher in SS from Suisun Bay. The ratio of oxidized glutathione (GSSG) to reduced glutathione (GSH) was greater in all species from Tomales Bay. The following significant relationships were found in one or more species with increasing hepatic Hg concentration: lower body, liver, and heart weights; decreased hepatic GSH concentration and G-6-PDH and GSH peroxidase activities; increased ratio of GSSG to GSH; and increased GSSG reductase activity. With increasing hepatic Se concentration, GSH peroxidase increased, but GSH decreased. It is concluded that measurement of associated enzymes in conjunction with thiol status may be a useful bioindicator to discriminate between Hg and Se effects. Concentrations of Hg and Se and the above variables affected have been associated with adverse effects on reproduction and neurological function in experimental studies with mallards.

Environmental Toxicology and Chemistry

Cyanobacteria reduce motility of quagga mussel (Driessena rostriformis bugensis) sperm

The temporal expansion of harmful algal blooms, primarily associated with cyanobacteria, may impact aquatic organisms at vulnerable life history stages. Broadcast spawning species release gametes into the water column for external fertilization, directly exposing sperm to potential aquatic stressors. To determine if cyanobacteria can disrupt reproduction in freshwater broadcast spawners, we evaluated sub‐lethal effects of cyanobacteria exposure on quagga mussel ( Dreissena rostriformis bugensis ) sperm. In laboratory studies, sperm were collected after inducing mussels to spawn using serotonin and exposed to 11 cultures of cyanobacteria including Anabaena flos‐aquae , Aphanizomenon flos‐aquae , Dolichospermum lemmermanii , Gloeotrichia echinulata , five cultures of Microcystis aeruginosa, M. wesenbergii , and Planktothrix suspensa . Sperm motility, using endpoints of cumulative distance traveled and mean velocity was calculated for a minimum of 10 individual sperm using a novel optical biotracking assay method. The distance and velocity at which sperm travelled decreased when exposed to Aphanizomenon flos‐aquae and two M. aeruginosa cultures. Our findings indicate that cyanobacteria impede the motility of quagga mussel sperm, which can potentially result in reproductive impairments to mussels, and potentially other broadcast spawning species.

Environmental Toxicology and Chemistry

Effects of DDT on bobwhite quail adrenal gland

A wide range of responses to sublethal levels of DDT exist, many of which are species specific and vary within each species depending upon age, sex, and physiological state. Sublethal levels of DDT do cause an increase in the adrenal cortical tissue of bobwhite quail, which may cause increased secretion of corticosteroids, and in turn affect reproduction. A delicate homeostatic balance exists within the avian endocrine system which may be disturbed by feeding sublethal levels of chlorinated hydrocarbon pesticides. This adverse effect on the endocrine system may cause subtle reproductive failures which go unnoticed until the population is greatly reduced.

Bulletin of Environmental Contamination and Toxico

Control of invasive sea lampreys using the piscicides TFM and niclosamide: Toxicology, successes & future prospects

The invasion of the Laurentian Great Lakes of North America by sea lampreys ( Petromyzon marinus ) in the early 20th century contributed to the depletion of commercial, recreational and culturally important fish populations, devastating the economies of communities that relied on the fishery. Sea lamprey populations were subsequently controlled using an aggressive integrated pest-management program which employed barriers and traps to prevent sea lamprey from migrating to their spawning grounds and the use of the piscicides (lampricides) 3-trifluoromethyl-4-nitrophenol (TFM) and niclosamide to eliminate larval sea lampreys from their nursery streams. Although sea lampreys have not been eradicated from the Great Lakes, populations have been suppressed to less than 10% of their peak numbers in the mid-1900s. The ongoing use of lampricides provides the foundation for sea lamprey control in the Great Lakes, one of the most successful invasive species control programs in the world. Yet, significant gaps remain in our understanding of how lampricides are taken-up and handled by sea lampreys, how lampricides exert their toxic effects, and how they adversely affect non-target invertebrate and vertebrates species. In this review we examine what has been learned about the uptake, handling and elimination, and the mode of TFM and niclosamide toxicity in lampreys and in non-target animals, particularly in the last 10 years. It is now clear that the mode of TFM toxicity is the same in non-target fishes and lampreys, in which TFM interferes with oxidative phosphorylation by the mitochondria leading to decreased ATP production. Vulnerability to TFM is related to abiotic factors such as water pH and alkalinity, which we propose changes the relative amounts of the bioavailable un-ionized form of TFM in the gill microenvironment. Niclosamide, which is also a molluscicide used to control snails in areas prone to schistosomiasis infections of humans, also likely works by uncoupling oxidative phosphorylation, but less is known about other aspects of its toxicology. The effects of TFM include reductions in energy stores, particularly glycogen and high energy phosphagens. However, non-target fishes readily recover from sub-lethal TFM exposure as demonstrated by the rapid restoration of energy stores and clearance of TFM. Although both TFM and niclosamide are non-persistent in the environment and critical for sea lamprey control, increasing public and institutional concerns about pesticides in the environment makes it imperative to explore other means of sea lamprey control. Accordingly, we also address possible “next-generation” strategies of sea lamprey control including genetic tools such as RNA interference and CRISPR-Cas9 to impair critical physiological processes (e.g. reproduction, digestion, metamorphosis) in lamprey, and the use of green chemistry to develop more environmentally benign chemical methods of sea lamprey control.

Great Lakes

Avian nesting success and diversity in conventionally and organically managed apple orchards

This study examines the effects of operational use of pesticides on avian species inhabiting apple orchards in Pennsylvania Mourning dove ( Zenaida macroura ) and American robin ( Turdus migratorius ) nests were monitored in three organic and three conventional apple orchards during 1990 and 1991 In 1991 we surveyed the avian communities of these orchards by using line transects Organophosphorus (OP) (e g, azinphos methyl, phosphamidon, parathion, dimethoate), carbamate (CA) (e g, methomyl, formetanate, oxamyl), and organochlorine (endosulfan) pesticides, known to be highly toxic to birds, were sprayed individually or in mixtures as part of routine pest management as many as 19 times during peaks in breeding activity Spray card tests revealed that OP pesticides were deposited on 86% of the nests in conventional orchards Daily survival rates (DSRs) for nests of both species were higher in the organic orchards than in the conventional orchards for 1991 and for years combined ( p < 0 05) Species diversity was greater in the organic orchards ( H = 2 43) than in the conventional orchards ( H = 1 79) Repeated applications of pesticides within the conventional orchards reduced the reproductive success of doves and robins and may have lowered avian species diversity compared with organic orchards

Pennsylvania

Hepatic element concentrations of lesser scaup (Aythya affinis) during spring migration in the upper Midwest

High concentrations of some hepatic elements might be contributing to the decline of the continental lesser scaup ( Aythya affinis ) population. We evaluated hepatic element concentrations of male and female lesser scaup collected from the upper Midwest (Iowa, Minnesota, North Dakota, and South Dakota) during the 2003 and 2004 spring migrations. We measured concentrations of 24 elements in livers of 117 lesser scaup. We found that only selenium concentrations were at levels (>3.0 μg/g wet weight [ww)]) proposed to adversely affect reproduction. Approximately 49% of females ( n = 61) had individual hepatic concentrations >3.0 μg/g ww selenium (Se). Our observed hepatic concentration of Se was similar to that reported in lesser scaup collected from the mid-continental United States but less than Se concentrations reported from the Great Lakes region. We found that the liver cadmium (Cd) concentration for males was significantly higher than that for females. Gender differences in hepatic Cd concentrations have not been previously reported for lesser scaup, but Cd is known to have negative impacts on male reproduction. Our results indicate that lesser scaup migrating through the upper Midwest in spring have elevated Se levels and that males carry a significantly greater Cd burden than females. Moreover, elemental concentrations might be high enough to affect reproduction in both male and female lesser scaup, but controlled laboratory studies are needed to adequately assess the effects of Se and Cd on lesser scaup reproduction.

Iowa, Minnesota, North Dakota, South Dakota

Orthene® toxicity to little brown bats (Myotis lucifugus): Acetylcholinesterase inhibition, coordination loss, and mortality

The 24-h LD50 of Orthener̀ (active ingredient acephate, acetylphosphoramidothioic acid o,s -dimethyl ester, CAS 30560-19–1) to little brown bats (Myotis lucifugus) was high (> 1,500 mg acephate/kg) and at least several times greater than the LD50 for mice (Mus musculus) (720 mg/kg). Twenty-four hours after dosing, all surviving mice appeared behaviorally normal, but 9 of 30 surviving bats could not right themselves when placed on their backs. When dead and incapacitated bats were combined to calculate an ED50 (median effective dose), the resultant estimate (687 mg/kg) did not differ ( p > 0.05) from the LD50 for mice. Serum cholinesterase (ChE) activity in control bats was 3.2 times greater than in mice. The relationship between this naturally high level of ChE and the relative tolerance of bats to organophosphorus insecticides is unexplained. Toxicity of Orthene was clearly less than that reported elsewhere for methyl parathion (phosphorothioic acid o,o -dimethyl o-[4-nitrophenyl] ester, CAS 298–00-0). This finding may be useful in selection of a chemical for agricultural use, but conclusions about the safety of Orthene to this bat species, or to others, must remain tentative until confirmed by studies under field conditions. Because bats are long-lived with low reproductive rates and slow recruitment, any additional mortality in the wild could be critical to population survival.

Environmental Toxicology and Chemistry

Reproductive responses of male fathead minnows exposed to wastewater treatment plant effluent, effluent treated with XAD8 resin, and an environmentally relevant mixture of alkylphenol compounds

On-site, continuous-flow experiments were conducted during August and October 2002 at a major metropolitan wastewater treatment plant (WWTP) to determine if effluent exposure induced endocrine disruption as manifested in the reproductive competence of sexually mature male fathead minnows ( Pimephales promelas ). The fathead minnows were exposed in parallel experiments to WWTP effluent and WWTP effluent treated with XAD8 macroreticular resin to remove the hydrophobic-neutral fraction which contained steroidal hormones, alkylphenolethoxylates (APEs), and other potential endocrine disrupting compounds (EDCs). The effluent composition varied on a temporal scale and the continuous-flow experiments captured the range of chemical variability that occurred during normal WWTP operations. Exposure to WWTP effluent resulted in vitellogenin induction in male fathead minnows, with greater response in October than in August. Concentrations of ammonia, APEs, 17β-estradiol, and other EDCs also were greater in October than in August, reflecting a change in effluent composition. In the October experiment, XAD8 treatment significantly reduced vitellogenin induction in the male fathead minnows relative to the untreated effluent, whereas in August, XAD8 treatment had little effect. During both experiments, XAD8 treatment removed greater than 90% of the APEs. Exposure of fish to a mixture of APEs similar in composition and concentration to the WWTP effluent, but prepared in groundwater and conducted at a separate facility, elicited vitellogenin induction during both experiments. There was a positive relation between vitellogenin induction and hepatosomatic index (HSI), but not gonadosomatic index (GSI), secondary sexual characteristics index (SSCI), or reproductive competency. In contrast to expectations, the GSI and SSCI increased in males exposed to WWTP effluent compared to groundwater controls. The GSI, SSCI, and reproductive competency were positively affected by XAD8 treatment of the WWTP effluent.

Aquatic Toxicology

Organochlorine contaminants and reproductive success of double-crested cormorants from Green Bay, Wisconsin, USA

In 1994 and 1995, nesting success of double-crested cormorants ( Phalacrocorax auritus ) was measured at Cat Island, in southern Green Bay, Lake Michigan, Wisconsin, USA. Sample eggs at pipping and unhatched eggs were collected and analyzed for organochlorines (including total polychlorinated biphenyls [PCBs] and DDE), hepatic microsomal ethoxyresorufin-O-dealkylase (EROD) activity in embryos, and eggshell thickness. Of 1,570 eggs laid, 32% did not hatch and 0.4% had deformed embryos. Of 632 chicks monitored from hatching to 12 d of age, 9% were missing or found dead; no deformities were observed. The PCB concentrations in sample eggs from clutches with deformed embryos (mean = 10.2 μg/g wet weight) and dead embryos (11.4 μg/g) were not significantly higher than concentrations in sample eggs from nests where all eggs hatched (12.1 μg/g). A logistic regression of hatching success versus DDE, dieldrin, and PCB concentrations in sibling eggs identified DDE and not dieldrin or PCBs as a significant risk factor. A logistic regression of hatching success versus DDE and eggshell thickness implicated DDE and not eggshell thickness as a significant risk factor. Even though the insecticide DDT was banned in the early 1970s, we suggest that DDE concentrations in double-crested cormorant eggs in Green Bay are still having an effect on reproduction in this species.

Wisconsin

Selenium and heavy metals in San Francisco Bay diving ducks

We analyzed for selenium (Se) and heavy metals in greater scaups ( Aythya marila ) and surf scoters ( Melanitta perspicillata ) collected from southern San Francisco Bay in March and April 1982. There were no differences (P > 0.05) between species for liver concentrations of silver (Ag), mercury (Hg), or lead (Pb). Copper (Cu) (P < 0.001) and zinc (Zn) (P < 0.01) levels were higher in scaups, whereas Se was higher in scoters (P < 0.001). Chromium (Cr) and nickel (Ni) occurred in < 50% of the samples, and there was no difference (P > 0.05) between the 2 species. The geometric mean cadmium (Cd) concentration in scoter kidneys (24.6 ppm, dry wt) was higher than in scaups (15.5 ppm) (0.1 > P > 0.05). Liver concentrations of Hg and Se were correlated (P < 0.01). The toxicological significance of some elements in these species is not known. However, Se levels in scoters (34.4 ppm, dry wt) were similar to those in livers of dabbling ducks (Anas spp.) in the nearby San Joaquin Valley where reproduction was impaired severely.

California

Contaminants in eggs of aquatic birds from the grasslands of central California

Eggs were collected from the Grasslands of western Merced County, California, during 1986 and 1987, and at the Mendota Wildlife Area in Fresno County in 1987, as part of a reproductive study of nesting ducks and shorebirds. The eggs were analyzed to evaluate the effects of contaminants in agricultural drainwater on avian reproduction. Agricultural drainwater was the major source of water for irrigation and fall flooding in the Grasslands until the fall of 1985 when drainwater was mostly replaced by uncontaminated water. Drainwater had not been used at Mendota. Concentrations of arsenic and boron in eggs from all sites were lower than those known to harm avian reproduction. However, mean concentrations of selenium, a trace element known to impair avian reproduction at high dietary levels, were higher in eggs from the Grasslands than in eggs from the uncontaminated site. Although some selenium concentrations were as high as those found to have caused death or deformities at other sites, reproductive impairment was not observed in any ducks or shorebirds nesting in the Grasslands in 1986 or 1987.

California

Perfluoroalkyl contaminant exposure in tree swallows nesting at Clarks Marsh, Oscoda, Michigan, USA

A site in north eastern Michigan, Oscoda Township, has some of the highest recorded exposure in birds to perfluorinated substances (PFASs) in the U.S. Some egg and plasma concentrations at that location exceeded the lowest reproductive effect threshold established for two avian laboratory species. The objectives of this study were to determine whether there were reproductive effects or physiological responses in a model bird species, the tree swallow (Tachycineta bicolor), associated with this extremely high exposure to PFASs. The lack of exposure above background to other contaminants at this site allowed for an assessment of PFAS effects without the complication that responses may be caused by other contaminants. A secondary objective was to determine the distribution of PFASs in multiple tissue types to better understand and interpret residues in different tissues. This can best be done at highly exposed locations where tissue concentrations would be expected to be above detectable levels if they are present in that tissue. There were no demonstrable effects of PFAS exposure on reproduction nor on most physiological responses.

Michigan