Influence of metal concentrations, percent salinity, and length of exposure on the metabolic rate of fathead minnows (Pimephales promelas)

Understanding the effects of chemical toxicants on energetic processes is an important aspect of ecotoxicology. However, the influence of toxicant concentration and time of exposure on metabolism in aquatic organisms is still poorly understood. The purpose of this investigation was to determine the influence of increasing levels of three stressors (Cu, Cd, percent salinity) and exposure time (24 h and 96 h) on the metabolic rate of fathead minnows (Pimephales promelas). In all 24-h exposures, there existed a threshold concentration, above which metabolic rate decreased significantly compared to the control and lower concentrations. In contrast, the metabolic rate of fish exposed for 96 h increased significantly in all concentrations compared to fish from the control. We suggest fathead minnows exhibit a consistent pattern of metabolic response to stressors, regardless of the physiological mechanisms involved, and that this response differs as a function of time of exposure.     

Comparison of ELISAs for detecting vitellogenin in the fathead minnow (Pimephales promelas)

Measurement of vitellogenin (VTG) concentrations in the fathead minnow (Pimephales promelas) is currently being considered and evaluated for screening of endocrine active substances. One of the proposed methods, an enzyme-linked immunosorbent assay (ELISA) based on VTG from carp (Cyprinus carpio), was recently evaluated in an inter-laboratory ring test using whole body homogenates from juvenile fathead minnows. The objective of the current study was to compare the performance of three different ELISAs for measuring fathead minnow VTG: (1) a heterologous carp VTG (cVTG) ELISA used in the ring test, (2) a homologous fathead minnow VTG (fVTG) ELISA, and (3) a hybrid ELISA with the antibody developed for cVTG, but using fVTG for coating the plates and preparing standard curves. VTG was measured in whole body homogenates from juvenile fathead minnows exposed to 17alpha-ethynylestradiol (EE(2); 10 ng/l) and whole body homogenates and plasma from adult fathead minnows exposed to 17beta-estradiol (E(2); 5 mg/kg; i.p.). The cVTG assay showed lower specificity for fathead minnow VTG in whole body homogenates and plasma from treated fish, compared to the fVTG assay. VTG concentrations in juvenile fathead minnow homogenates from the EE(2)-exposed group were approximately 50-fold higher when measured using the fVTG method compared to the cVTG method. Use of the homologous fVTG in the hybrid cVTG assay yielded VTG concentrations in the range of the fVTG assay but the low specificity persisted. The homologous fVTG assay is recommended to achieve accurate quantification of VTG levels in fathead minnows.     

Profiling lipid metabolites yields unique information on sex- and time-dependent responses of fathead minnows (Pimephales promelas) exposed to 17α-ethynylestradiol

Alterations in hepatic lipid profiles of fathead minnows (FHM) exposed to the synthetic estrogen 17α-ethynylestradiol (EE2) were determined using ¹H-NMR spectroscopy-based metabolite profiling. The exposures were conducted using either 10 ng/l or 100 ng/l EE2 via a continuous flow water delivery system. Livers were collected at 1, 4, and 8 days of the exposure and 8 days after the chemical was removed from the water (i.e. an 8 day depuration). The exposure resulted in a number of sex-specific changes in lipid profiles that were also highly time dependent. Those metabolites most affected by exposure included phosphatidylcholine, diglycerides, triglycerides and cholesterol. In addition, changes in the length and degree of unsaturation of hepatic fatty acids were observed. Lipid profiles in plasma for fish collected on the 4th day of exposure were also analyzed in order to provide further insights into changes observed in hepatic metabolite changes. Using validated partial least-squares discriminant analysis (PLS-DA), the response trajectories of the male liver lipid profiles at both exposure concentrations were compared. This analysis indicated that the males exposed to the low concentration of EE2 (10 ng/l) were largely able to recover from the exposure once the chemical was removed from the water. Conversely, the males exposed to the high concentration (100 ng/l) did not appear to recover from the exposure despite the 8 day depuration.     

Temporal characterization of growth of fathead minnow (Pimephales promelas) larvae during sublethal hydrogen cyanide exposure

Growth of fathead minnow yolk sac larvae was characterized from changes in dry weight and total content and concentrations of RNA, DNA and protein in fish exposed to a sublethal level of HCN (58 micrograms/l) and in age matched controls. Cyanide toxicosis occurred within 24 hr of exposure as evidenced by significant reductions in protein and RNA content and RNA/DNA ratio of larvae. After 96 hr exposure to HCN, larvae exhibited the same growth rate and protein synthetic rate (RNA/DNA) as control fish. HCN toxicosis and recovery is rapid and at least partial tolerance to HCN develops within 96 hr of exposure in larval fathead minnows.     

Foraging Trade-offs in Fathead Minnows (Pimephales promelas,Osteichthyes, Cyprinidae): Acquired Predator Recognition in the Absence of an Alarm Response

Pike-naive fathead minnows (Pimephales promelas) were fed ad libitum or deprived of food for 12, 24, or 48 h and then exposed to either conspecific alarm pheromone or distilled water and the odour of a predatory northern pike (Esox lucius). Minnows fed ad libitum or deprived for 12 h showed a stereotypic alarm response to the alarm pheromone (increased time under cover objects and increased occurrence of dashing and freezing behaviour); those deprived of food for 24 h showed a significantly reduced alarm response, while those deprived of food for 48 h did not differ significantly from the minnows exposed to a distilled water control. Upon subsequent testing in an Opto-Varimex activity meter, all groups initially exposed to alarm pheromone and pike odour exhibited an alarm response when exposed to pike odour alone. Those initially conditioned with distilled water and pike odour did nor show an alarm response to pike odour alone. These results demonstrate that there exists a significant trade-off between hunger level and predator-avoidance behaviour in fathead minnows and that minnows can learn the chemical cues of a predatory northern pike through association with alarm pheromone even in the absence of an observable alarm response.     

Influence of dissolved organic matter on copper binding, and calcium on cadmium binding, by gills of rainbow trout

Complexation of Cu by 5 mg Cl⁻¹ dissolved organic matter (DOM) from a marsh kept Cu from binding to gills of small rainbow trout Oncorhynchus mykiss in 9-day exposures to 0.5 μM Cu in soft water. The protective effect of DOM occurs because the formation of Cu-DOM complexes reduces the amount of free Cu in the water, so the disruptive effects of Cu on ionoregulation, such as inhibited Na uptake, cannot develop. The Cu-DOM complexes themselves do not bind to the gills. Calcium (1100 μm) reduced the accumulation of Cd by trout gills in short, 2-h exposures through competition for gill binding sites but not over longer, 7-day exposures to 0–14 μM Cd. However, the protective effect of Ca against Cd toxicity persisted throughout the longer experiment, likely due to the decrease in the electrochemical gradient for diffusive loss of Ca from the fish to the water. Rainbow trout and fathead minnows Pimephales promelas accumulated Cu and Cd on their gills in a similar manner; thus, binding constants for metal-gill interactions determined for one species of fish can be generalized to other fish species. When literature binding constants determined for fathead minnows were applied to our studies with rainbow trout, computer modelling of Cu-gill and Cu-DOM interactions simulated our results well. In contrast Cd-gill and Ca-gill modelling predicted the initial competitive effect of Ca against Cd accumulation by trout gills, but did not predict the longer-term accumulation of Cd by trout gills.     

The role of experience and chemical alarm signalling in predator recognition by fathead minnows, Pimephales promelas

Young-of-the-year, predator-naive fathead minnows, Pimephales promelas , from a pikesympatric population did not respond to chemical stimuli from northern pike, Esox Indus , while wild-caught fish of the same age and size did. These results suggest that chemical predator recognition is a result of previous experience and not genetic factors, Wild young-of-the-year minnows responded to pike odour with a response intensity that was similar to that of older fish, demonstrating that the ability to recognize predators is learned within the first year. The intensity of response of wild minnows which had been maintained in a predator free environment for 1 year was similar to that of recently caught minnows of the same age, suggesting that reinforcement was not required for predator recognition to be retained. Naive minnows that were exposed simultaneously to chemical stimuli from pike (a neutral stimulus) and minnow alarm substance exhibited a fright response upon subsequent exposure to the pike stimulus alone. Predator-naive minnows exposed simultaneously to chemical stimuli from pike and glass-distilled water did not exhibit a fright response to the pike stimulus alone. These results demonstrate that fathead minnows can acquire predator recognition through releaserinduced recognition learning, thus confirming a known mechanism through which alarm substance may benefit the receivers of an alarm signal.     

Serum-thyroxine levels in microwave-exposed rats

The nature of the response of the thyroid gland in animals exposed to microwave irradiation is controversial. An enlarged thyroid and an increase of radioiodine uptake in microwave workers have been reported. Absence of thyroid disorders has also been reported in other exposed populations. Animal experimentation has contributed to the controversy because both increased and decreased thyroid functions have been reported. The thyroxine concentration in rats as representative of thyroid function in animals exposed to 2.45-GHz, 120-Hz amplitude-modulated microwaves has been studied. Comparison was made between thyroxine concentrations in microwave- and sham-exposed rats by Student's t test. After a 1-hr exposure, an increased thyroxine concentration was found in rats exposed at 40 and 70 mW/cm2, but not at 1, 5, 10, 20, 50, or 60 mW/cm2. After a 2-hr exposure, increased thyroxine concentration was noted in rats exposed at 25, 30, and 40 mW/cm2, but not at 1, 5, 10, and 20 mW/cm2. After a 4-hr exposure, thyroxine concentration increased in rats exposed at 1 mW/cm2 and decreased in rats exposed at 20 mW/cm2; but changes were not noted at 5 or 10 mW/cm2. Other experiments included animals that were exposed once for 4 hr (0.1, 1, 10, 25, and 40 mW/cm2), sampled 24 hr after a 4-hr exposure (0.1, 1, 10, 25, and 40 mW/cm2), or exposed for 4 hr 3 times (1, 10, 20, 30, 40, and 55 mW/cm2) and 10 times (1, 10, 20, 25, 30, and 40 mW/cm2), to evaluate the consistency of the thyroxine response. None of the rats in these experiments displayed any alteration of thyroxine concentration, except that decreased thyroxine was noted in rats exposed at 40 mW/cm2 for the third time. These studies covered a long time span; rats from two commercial sources (BS and CR) were used and subjected to different numbers of exposures, and therefore these data were evaluated for their stability. Two factors could influence the result significantly, i.e., source of animal and number of sham exposures. Rats used in the 2-hr exposures were from two different commercial sources; rats from CR had a higher (but normal) thyroxine concentration than did rats from BS. Therefore the data of these animals were separated by commercial source for reevaluation. Instead of increased thyroxine concentration in rats exposed at 25, 30, and 40 mW/cm2, changes were not noted in any microwave-exposed rats. The influence of sham exposure revealed that appropriate concurrent control and specification of animal source are needed in longitudinal studies.(ABSTRACT TRUNCATED AT 400 WORDS)     

Physiological, anatomic and behavioural changes in the respiratory system of Salmo gairdneri Rich. On acute and chronic exposure to chlorothalonil

1. Various changes in anatomical, physiological and behavioural aspects of the respiratory system of Salmo gairdneri on exposure to chlorothalonil (TCIN) were examined, along with the relationship between LC50 response and oxygen concentration. 2. The TCIN LC50 was significantly lower at 5 mg/l oxygen concentration than at 8 mg/l. 3. Haematocrit was depressed on exposure to TCIN, whereas exposure to pentachlorophenol and lindane raised the haematocrit. Muscle lactate was not elevated in acutely exposed fish. 4. Ventilatory frequency was raised above 30 micrograms/l TCIN. 5. Gills of fish exposed to 2.0 micrograms/l TCIN for 24 days had reduced lamellar diffusive capacity, primarily caused by an increase in blood barrier thickness.     

Selenium compounds in the fathead minnow (Pimephales promelas)--I. Uptake, distribution, and elimination of orally administered selenate, selenite and l-selenomethionine

Treatment of fathead minnows (Pimephales promelas) with either [75Se]selenate, -selenite or -l-selenomethionine by gavage at 20 ng Se/g resulted in organ uptake and early distribution patterns which differed significantly between compounds. The greatest differences in uptake between compounds was observed in liver tissue which accumulated much less [75Se]selenate than either selenite or l-selenomethionine. The 75Se burdens and relative distribution among the various organs were nearly identical during the elimination phase for [75Se]selenate and -selenite. This suggests that selenium derived from these compounds converge to a common metabolic pool. The whole body T1/2, rate of 75Se uptake and magnitude of 75Se accumulation were generally greater for [75Se]selenomethionine than the inorganic forms. Selenium-75 was present in the bile following the oral administration of each compound. The partitioning of selenate and selenite into the plasma and cellular fraction of blood differs with both the compound and time following exposure.     

Assessing the efficacy of corn-based bait containing antimycin-a to control common carp populations using laboratory and pond experiments

Strategic use of oral toxicants could allow for practical and sustainable control schemes for the invasive common carp (Cyprinus carpio, or ‘carp’) if a toxicant selectively targeted carp and not native species. In this study, we incorporated antimycin-a (ANT-A), a known fish toxicant, into a corn-based bait and conducted a series of experiments to determine its toxicity, leaching rate, and species-specificity. Our results showed that ANT-A was lethal to carp at doses ≥ 4 mg/kg and that the amount of ANT-A that leached out of the bait in 72 h was not lethal to carp or bluegill (Lepomis macrochirus). Species-specificity trials were conducted in 227 L tanks, in which carp were stocked with three native species representing families that occur sympatrically with carp in our study region: the fathead minnow (Pimephales promelas), yellow perch (Perca flavescens) and bluegill. These trials showed high mortality of carp (46%) and fathead minnows (76%) but no significant mortality of perch or bluegill. Finally, a pond study, which used the same species composition except for fathead minnows, resulted in 37% morality among adult carp and no mortality among perch or bluegill. Our results suggest that corn-based bait that contains ANT-A could be used to selectively control carp in ecosystems dominated by percids or centrarchids, such as lakes across the Great Plains ecoregion of North America, where carp are especially problematic.     

Predicting Fecundity of Fathead Minnows (Pimephales promelas) Exposed to Endocrine-Disrupting Chemicals Using a MATLAB?-Based Model of Oocyte Growth Dynamics

Fish spawning is often used as an integrated measure of reproductive toxicity, and an indicator of aquatic ecosystem health in the context of forecasting potential population-level effects considered important for ecological risk assessment. Consequently, there is a need for flexible, widely-applicable, biologically-based models that can predict changes in fecundity in response to chemical exposures, based on readily measured biochemical endpoints, such as plasma vitellogenin (VTG) concentrations, as input parameters. Herein we describe a MATLAB^® version of an oocyte growth dynamics model for fathead minnows (Pimephales promelas) with a graphical user interface based upon a previously published model developed with MCSim software and evaluated with data from fathead minnows exposed to an androgenic chemical, 17β-trenbolone. We extended the evaluation of our new model to include six chemicals that inhibit enzymes involved in steroid biosynthesis: fadrozole, ketoconazole, propiconazole, prochloraz, fenarimol, and trilostane. In addition, for unexposed fathead minnows from group spawning design studies, and those exposed to the six chemicals, we evaluated whether the model is capable of predicting the average number of eggs per spawn and the average number of spawns per female, which was not evaluated previously. The new model is significantly improved in terms of ease of use, platform independence, and utility for providing output in a format that can be used as input into a population dynamics model. Model-predicted minimum and maximum cumulative fecundity over time encompassed the observed data for fadrozole and most propiconazole, prochloraz, fenarimol and trilostane treatments, but did not consistently replicate results from ketoconazole treatments. For average fecundity (eggs•female^-1•day^-1), eggs per spawn, and the number of spawns per female, the range of model-predicted values generally encompassed the experimentally observed values. Overall, we found that the model predicts reproduction metrics robustly and its predictions capture the variability in the experimentally observed data.     

Effects of exposure to predatory cues on territorial behaviour of male fathead minnows

We conducted a laboratory study to determine if male fathead minnows, Pimephales promelas, altered their territorial behaviour associated with reproduction in response to combinations of visual and chemical cues from northern pike, Esox lucius. We introduced the following stimuli to a territorial male: a brick (control), fathead minnow alarm pheromone, a pike fed brook stickleback, Culea inconstans, or a pike fed fathead minnow. The territorial behaviour of males did not change when the control was added. Male minnows experiencing threat from pike fed stickleback significantly reduced the frequency at which they performed three territorial behaviours, but, within 12 h, had returned to pre-exposure activity levels. Male minnows subjected to alarm pheromone alone and to pike fed fathead minnow significantly reduced their territorial behaviour, abandoned their nests, and did not return to pre-exposure levels of activity after 24 h. We suggest that because risk of predation triggers prolonged decreases in territorial defense, it may affect competition between nesting males and female mate choice. We conclude that fathead minnows can assess the severity of predatory threat and adjust their reproductive behaviour accordingly.     

Oxygen consumption by eggs and larvae of striped mullet, Mugil cephalus, in relation to development, salinity and temperature

Oxygen consumption rates during embryonic and the first 38 days of larval development of the striped mullet were measured at 24° C by differential respirometry. Measurements were obtained at the blastula, gastrula and four embryonic stages, and at the yolk-sac, preflexion, flexion and post-flexion larval stages.
Oxygen uptake rates of eggs increased linearly from 0.024 μl O₂ per egg h^-1 (0·323 μl O₂ mg^-1 dry wt h^-1) by blastulae to 0·177 μlO₂ per egg h^-1 (2·516 μlO₂mg ¹dry wth^-1) by embryos prior to hatching. Respiration rates did not vary significantly among four salinities (20,25, 30, 35%₀).
Larval oxygen consumption increased in a curvilinear manner from 0·243 μl O₂ per larva h^-1 shortly after hatching to 18·880 μl O₂ per larva h^-1 on day 38. Oxygen consumption varied in direct proportion to dry weight. Mass-specific oxygen consumption rates of preflexion, flexion, and postflexion larvae did not change with age (10·838 μl O₂ mg ¹dry wt h^-1).
Larval oxygen consumption rates did not vary significantly among salinities 10–35%. Acute temperature increases elicited significant increases in oxygen consumption, these being relatively greater in yolk-sac larvae ( Q₁₀ = 2·75) than in postflexion larvae ( Q₁₀ = 1·40).     

虎斑乌贼的胚胎耗氧率

为了探究虎斑乌贼胚胎不同发育时期的耗氧率变化和几种生态因子对胚胎发育过程耗氧率的影响,试验采用封闭静水装置,对不同发育时期(12期)的耗氧率进行测定,并研究不同盐度(21、24、27、30、33)、温度(18、21、24、27、30 ℃)和pH(7.0、7.5、8.0、8.5、9.0)对胚胎4个主要发育时期(受精卵期、原肠胚期、器官形成期和内骨骼形成期)耗氧率的影响.结果表明: 胚胎各个发育时期耗氧率不同,随着发育的进程而增大,受精卵期为0.082 mg·(100 eggs)^-1·h^-1,而到原肠胚期的耗氧率显著升高,为0.279 mg·(100 eggs)^-1·h^-1,到孵化期时,耗氧率达到1.367 mg·(100 eggs)^-1·h^-1;盐度对器官形成期和内骨骼形成期的耗氧率均有显著影响(P<0.05),对受精卵期和原肠胚期影响不显著(P>0.05),当盐度为30时,4个发育时期耗氧率均达到最大值,分别为0.082、0.200、0.768和1.301 mg·(100 eggs)^-1·h^-1;温度对原肠胚期、器官形成期和内骨骼形成期的耗氧率有显著影响(P<0.05),对受精卵期无显著性影响(P>0.05),在27 ℃时,胚胎4个发育时期均达到最大值,分别为0.082、0.286、0.806和1.338 mg·(100 eggs)^-1·h^-1;而pH对4个发育时期的耗氧率均无显著性影响(P>0.05),受精卵期在pH 8.0时达到最大值,为0.116 mg·(100 eggs)^-1·h^-1,原肠胚期、器官形成期、内骨骼形成期在pH 8.5时达到最大值,分别为0.281 、0.799和1.130 mg·(100 eggs)^-1·h^-1.     

虎斑乌贼的胚胎耗氧率

为了探究虎斑乌贼胚胎不同发育时期的耗氧率变化和几种生态因子对胚胎发育过程耗氧率的影响,试验采用封闭静水装置,对不同发育时期(12期)的耗氧率进行测定,并研究不同盐度(21、24、27、30、33)、温度(18、21、24、27、30 ℃)和pH(7.0、7.5、8.0、8.5、9.0)对胚胎4个主要发育时期(受精卵期、原肠胚期、器官形成期和内骨骼形成期)耗氧率的影响.结果表明: 胚胎各个发育时期耗氧率不同,随着发育的进程而增大,受精卵期为0.082 mg·(100 eggs)^-1·h^-1,而到原肠胚期的耗氧率显著升高,为0.279 mg·(100 eggs)^-1·h^-1,到孵化期时,耗氧率达到1.367 mg·(100 eggs)^-1·h^-1;盐度对器官形成期和内骨骼形成期的耗氧率均有显著影响(P<0.05),对受精卵期和原肠胚期影响不显著(P>0.05),当盐度为30时,4个发育时期耗氧率均达到最大值,分别为0.082、0.200、0.768和1.301 mg·(100 eggs)^-1·h^-1;温度对原肠胚期、器官形成期和内骨骼形成期的耗氧率有显著影响(P<0.05),对受精卵期无显著性影响(P>0.05),在27 ℃时,胚胎4个发育时期均达到最大值,分别为0.082、0.286、0.806和1.338 mg·(100 eggs)^-1·h^-1;而pH对4个发育时期的耗氧率均无显著性影响(P>0.05),受精卵期在pH 8.0时达到最大值,为0.116 mg·(100 eggs)^-1·h^-1,原肠胚期、器官形成期、内骨骼形成期在pH 8.5时达到最大值,分别为0.281 、0.799和1.130 mg·(100 eggs)^-1·h^-1.     

Variation in tolerance to hypoxia in a predator and prey species: an ecological advantage of being small?

Three physiological variables, haematocrit, haemoglobin concentration and ventilation frequency, were measured to test how fathead minnows Pimephales promelas and small and large yellow perch Perca flavescens responded to three different dissolved oxygen concentrations. All fish were monitored continuously for any indications of stress in response to these manipulations. Within and between species, smaller individuals were the most tolerant of hypoxic environments. A species effect, however, did contribute to this observation, with fathead minnows being more tolerant of hypoxic environments than similar-sized yellow perch. In aquatic ecosystems where smaller fishes are more tolerant to hypoxia than their larger predators, hypoxic environments may have the potential to act as a refuge from predators.     

浮选剂丁基黄原酸钠对草鱼早期发育阶段的毒性效应

丁基黄原酸钠是一种金属矿的浮选剂.作者研究了它对草鱼早期发育阶段(孵化、存活、生长、畸形等)的毒性效应.结果表明,丁基黄原酸钠对草鱼胚胎96h的半致死浓度为17.8mg/L,致畸半效应浓度为8.85mg/L,当浓度为24mg/L时,其胚胎的孵化率仅为38.8%,对生长有影响的浓度为5.6-10.0mg/L,浓度在5.6mg/L就可诱发畸形.丁基黄原酸钠对草鱼胚胎具有强烈的致崎作用,崎形的主要症状为弯体和体表瘤状赘生物.       

Natural disturbance and life history: consequences of winterkill on fathead minnow in boreal lakes

Age, growth and reproductive characteristics of fathead minnow Pimephales promelas populations inhabiting four lakes that varied in the extent and frequency of winterkill were studied in the boreal region of western Canada. The lifespan of fathead minnows inhabiting lakes prone to winterkill was 1–2 years shorter than those in less disturbed lakes. In populations prone to winterkill, fish displayed faster growth rates and grew to a larger size‐at‐age, particularly during the first year of life. Although lower population densities in winterkill lakes probably contributed to this increased growth, adults in these populations tended to spawn earlier in the season than the smaller adults in more stable populations. Fathead minnows in lakes prone to winterkill also matured at an earlier age and allocated a greater proportion of their body mass to gonads than conspecifics in the more benign, stable lakes. These trends are consistent with predictions for organisms in variable, unpredictable environments and, because fathead minnows are tolerant to a wide range of environmental conditions, suggest that variation in life‐history traits among populations is probably a product of both selection and phenotypic plasticity.