Disturbance cues function as a background risk cue but not as an associative learning cue in tadpoles.

Chemical information has an important role in the sensory ecology of aquatic species. For aquatic prey, chemical cues are a vital source of information related to predator avoidance and risk assessment. For instance, alarm cues are released by prey that have been injured by predators. In addition to providing accurate information about current risk, repeated exposure to alarm cues can elicit a fear response to novel stimuli (neophobia) in prey. Another source of chemical information is disturbance cues, released by prey that have been disturbed or harassed (but not injured) by a predator. While disturbance cues have received much less attention than alarm cues, they appear to be useful as an early warning signal of predation risk and have the potential to be used as a priming cue for learning. In this study, we used wood frog (Lithobates sylvaticus) tadpoles to test whether repeated exposure to disturbance cues during the embryonic stage can induce neophobic behaviour. Three weeks following repeated exposure to disturbance cues, tadpoles showed reduced activity when exposed to a novel odour, but they no longer displayed an antipredator response to disturbance cues. In a second experiment, we found that tadpoles failed to learn that a novel odour was dangerous following a pairing with disturbance cues, whereas alarm cues facilitated such learning. Our results add to the growing body of information about disturbance cues and provide evidence of their function as an embryonic risk cue but not as an associative learning cue.

Turnover of hydrogen isotopes in lake sturgeon blood: implications for tracking movements of wild populations.

Naturally occurring deuterium ((2)H) in biota can be used to trace movement, migration and geographic origin of a range of organisms. However, to evaluate movements of animals using δ(2)H measurements of tissues, it is necessary to establish the turnover time of (2)H in the tissues and the extent of isotopic discrimination from different environmental (2)H sources to those tissues. We investigated the turnover of (2)H in lake sturgeon (Acipenser fulvescens) blood by manipulating both environmental water δ(2)H and diet δ(2)H over a four-month period. The half-life of deuterium in lake sturgeon blood was 37.9 days after an increase in the environmental water δ(2)H of +714 ‰. However, no clear turnover in blood (2)H occurred over the same period in a separate trial following a change of -63.8 ‰ or +94.2 ‰ in diet. These findings suggest that environmental water (2)H exchanges much faster with blood than diets and that blood δ(2)H values can be used to trace movements of sturgeon and other fish moving among isotopically distinct waters.

Sub-lethal increases in salinity affect reproduction in fathead minnows.

Salinization poses a threat to many inland aquatic ecosystems, especially in areas where natural processes are compounded by anthropogenic salinization. Though physiological survival can be a challenge for stenohaline freshwater fishes facing increasing salinity, it is important to note that essential and complex activities such as reproduction may be affected well below physiological tolerance limits. Here, we exposed fathead minnows (Pimephales promelas) to four levels of salinity in order to assess any impacts on several egg production and behavioral endpoints. We found significant reductions in total eggs produced, percent fertilization, number of spawning days, clutch size, total time males spent in the nest, and duration of nest care events. Our data demonstrate that salinization can have negative effects on critical reproductive endpoints.

Investigating the link between pulp mill effluent and endocrine disruption: attempts to explain the presence of intersex fish in the Wabigoon River, Ontario, Canada.

The ability of some pulp mill effluents (PME) to act as reproductive and endocrine disrupters in fish is well documented in the literature. However, changes are not always consistent with regard to species, gender, hormones, or reproductive effects. In the present study, the presence of the first intersexed fish that, to our knowledge, has been found in a Canadian river exposed to PME, is reported. A field survey of the Wabigoon River near Dryden, Ontario, in the fall of 2000 found intersexed walleye (Sander vitreus vitreus) with significantly altered hormone levels and reduced gonad size. The Wabigoon River receives discharge from a bleached kraft pulp and paper mill and a municipal wastewater (MWW) plant. It also has historical sediment contamination (wood fiber mats) contributing to extended periods of low dissolved oxygen under low flow, drought conditions. A mesocosm-based partial life cycle test exposing fathead minnows (Pimephales promelas) to reference water, 20% effluent volume to river volume (v/v), 40% (v/v), or 60% (v/v) PME as well as a field survey of the walleye in the Wabigoon River were conducted. The only change in our mesocosm exposure was a decrease in testosterone in males with increasing effluent concentration and vitellogenin induction in males exposed to 60% (v/v) effluent. These results did not reflect the magnitude of endocrine disruption seen in the wild fish survey. Several hypotheses that may explain these discrepancies are proposed. Specifically, evidence is offered from published studies indicating that either hypoxia or MWW, alone or in combination with PME, may explain the discrepancy between our field experiment and the wild fish survey. The present study illustrates the complexities of multistressor receiving environments and the need for the development of cumulative effects assessment approaches.

Sleep deprivation can inhibit adult hippocampal neurogenesis independent of adrenal stress hormones.

Sleep deprivation (SD) can suppress cell proliferation in the hippocampal dentate gyrus of adult male rodents, suggesting that sleep may contribute to hippocampal functions by promoting neurogenesis. However, suppression of cell proliferation in rats by the platform-over-water SD method has been attributed to elevated corticosterone (Cort), a potent inhibitor of cell proliferation and nonspecific correlate of this procedure. We report here results that do not support this conclusion. Intact and adrenalectomized (ADX) male rats were subjected to a 96-h SD using multiple- and single-platform methods. New cells were identified by immunoreactivity for 5-bromo-2'-deoxyuridine (BrdU) or Ki67 and new neurons by immunoreactivity for BrdU and doublecortin. EEG recordings confirmed a 95% deprivation of rapid eye movement (REM) sleep and a 40% decrease of non-REM sleep. Cell proliferation in the dentate gyrus was suppressed by up to 50% in sleep-deprived rats relative to apparatus control or home cage control rats. This effect was also observed in ADX rats receiving continuous low-dose Cort replacement via subcutaneous minipumps but not in ADX rats receiving Cort replacement via drinking water. In these latter rats, Cort intake via water was reduced by 60% during SD; upregulation of cell proliferation by reduced Cort intake may obscure inhibitory effects of sleep loss on cell proliferation. SD had no effect on the percentage of new cells expressing a neuronal phenotype. These results demonstrate that the Cort replacement method is critical for detecting an effect of SD on cell proliferation and support a significant role for sleep in adult neurogenesis.

Epidermal 'alarm substance' cells of fishes maintained by non-alarm functions: possible defence against pathogens, parasites and UVB radiation.

Many fishes possess specialized epidermal cells that are ruptured by the teeth of predators, thus reliably indicating the presence of an actively foraging predator. Understanding the evolution of these cells has intrigued evolutionary ecologists because the release of these alarm chemicals is not voluntary. Here, we show that predation pressure does not influence alarm cell production in fishes. Alarm cell production is stimulated by exposure to skin-penetrating pathogens (water moulds: Saprolegnia ferax and Saprolegnia parasitica), skin-penetrating parasites (larval trematodes: Teleorchis sp. and Uvulifer sp.) and correlated with exposure to UV radiation. Suppression of the immune system with environmentally relevant levels of Cd inhibits alarm cell production of fishes challenged with Saprolegnia. These data are the first evidence that alarm substance cells have an immune function against ubiquitous environmental challenges to epidermal integrity. Our results indicate that these specialized cells arose and are maintained by natural selection owing to selfish benefits unrelated to predator-prey interactions. Cell contents released when these cells are damaged in predator attacks have secondarily acquired an ecological role as alarm cues because selection favours receivers to detect and respond adaptively to public information about predation.

Modafinil [2-[(diphenylmethyl)sulfinyl]acetamide] and circadian rhythms in syrian hamsters: assessment of the chronobiotic potential of a novel alerting compound.

In Syrian hamsters, behavioral procedures for inducing arousal (e.g., running in a novel wheel or gentle handling) can shift circadian rhythms when applied during the usual sleep period ("subjective day") and can attenuate phase shifts to light during the active period ("subjective night"). This raises the possibility that drugs that affect behavioral state may have "chronobiotic" potential. We characterized the effects of modafinil (2-[(diphenylmethyl)sulfinyl]acetamide), an atypical alerting compound, on circadian rhythms in male Syrian hamsters. Electroencephalogram recordings and video observations confirmed that modafinil dose dependently increases wakefulness at the expense of slow-wave and paradoxical sleep with no increase in locomotor activity per unit of time awake. Despite inducing arousal, modafinil at these doses (150 or 300 mg/kg), administered in the subjective day or early or late in the subjective night, did not perturb circadian phase. Modafinil (300 mg/kg) also had no effect on phase shifts to light exposure either early or late in the night and did not alter the size of phase shifts induced by running in a novel wheel for 3 h during the mid-day. Modafinil (300 mg/kg) did, however, decrease by approximately 50% the amount of novel wheel-stimulated running, moving leftward the dose-response relation between wheel revolutions and shift magnitude. These results indicate that, in Syrian hamsters, modafinil alone has no significant chronobiotic efficacy. Nevertheless, this agent may increase the sensitivity of the circadian pacemaker to nonphotic stimuli and may thus have some potential as a tool for promoting clock resetting in combination with behavioral strategies.

Microinjection of neostigmine into the pontine reticular formation of the mouse: further evaluation of a proposed REM sleep enhancement technique.

Microinjections of cholinergic agonists into the pontine reticular formation (PRF) powerfully induce rapid eye movement sleep (REMS) in cats but have comparatively weaker effects in rats. Recently, the cholinomimetic neostigmine has been reported to strongly enhance REMS following microinjection into the PRF of the mouse. That study used behavioral assessments of locomotion in lieu of electrophysiological measures of muscle tone to identify REMS. We sought to confirm that the behavioral state induced in mice by PRF injections of neostigmine meets standard electroencephalogram (EEG) and electromyogram (EMG) criteria for defining REMS. Cortical EEG, nuchal muscle EMG, and PGO waves were recorded from male C57BL/6N mice with chronic indwelling cannulae for the delivery of neostigmine to the PRF. Recordings were made during midday following injections of neostigmine (8.8 mM, 50 nl), 2 h after lights on (LD 12:12). Neostigmine induced a behavioral state characterized by low amplitude, highly desynchronized cortical EEG with little theta, no PGO waves, and a sustained high muscle tone. Behavioral states meeting standard criteria for slow-wave sleep (SWS) and REMS were significantly suppressed compared to baseline recordings, and REMS onset was delayed by 3 h. Consistent with earlier reports, neostigmine did strongly suppress locomotor activity in open field tests and in the home cage. Due to the failure to meet criteria for defining REMS, we conclude that neostigmine microinjection into the PRF of the mouse induces an abnormal waking state rather than REMS.

Rapid eye movement sleep induction by microinjection of the GABA-A antagonist bicuculline into the dorsal subcoeruleus area of the rat.

In cats, rapid eye movement sleep (REMS) can be induced rapidly and reliably by injections of the cholinergic agonist carbachol into the anterodorsal pontine tegmentum, also recognized as the perilocus coeruleus alpha, and designated the REMS Induction Zone (RIZ). In rats, the RIZ has been ascribed to a much larger and more ventral region within the entire oral pontine reticular formation. However, carbachol injections throughout this area produce only small, unreliable, and long latency REMS enhancements. The present study investigated whether REMS induction in the rat is possible by microinjection into the dorsal subcoeruleus nucleus (SubCD), a region with similarities to the cat RIZ. In freely moving unanaesthetized rats, microinjection of the GABA-A antagonist bicuculline significantly increased the amount and reduced the latency to REMS during a 2-h recording in the mid-light period. However, at effective doses, bicuculline usually also produced intermittent ipsiversive circling behavior that disrupted REMS maintenance. Attempts at eliminating this side-effect by: (i) coinjection of bicuculline with the NMDA antagonist, APV, (ii) lower bicuculline doses, or (iii) injection of the GABA-B antagonist, phaclofen, were unsuccessful. Other drugs injected into this area did not induce REMS; these included carbachol, the acetylcholinesterase inhibitor neostigmine, the glutamate agonist kainate, and vasopressin. In the rat, the SubCD is a highly sensitive region for both REMS induction and locomotor effects.