Development and environmental application of a genus-specific quantitative PCR approach for Pseudo-nitzschia species

Quantitative polymerase chain reaction (qPCR) for the identification and quantification of microbes has become a common tool for the study of harmful algal blooms (HABs). We developed a qPCR method for the diatom genus Pseudo-nitzschia. Several species of this genus form toxic blooms through the production of the neurotoxin domoic acid (DA). Outbreaks of toxicity attributed to DA along the US west coast have caused sickness and death of marine mammals and seabirds through food web contamination. The method developed here quantifies Pseudo-nitzschia spp. at low abundances in natural samples, thereby, providing a method to improve our understanding of the environmental conditions leading to blooms of these species. This has been accomplished previously by techniques for identification and quantification that are slow and laborious compared to qPCR. The approach was successfully tested and validated using eight species of Pseudo-nitzschia and 33 non-target organisms and employed to follow local bloom dynamics.     

Post-larval development of two intertidal barnacles at elevated CO2 and temperature

Ocean acidification and global warming are occurring concomitantly, yet few studies have investigated how organisms will respond to increases in both temperature and CO₂. Intertidal microcosms were used to examine growth, shell mineralogy and survival of two intertidal barnacle post-larvae, Semibalanus balanoides and Elminius modestus, at two temperatures (14 and 19°C) and two CO₂ concentrations (380 and 1,000 ppm), fed with a mixed diatom-flagellate diet at 15,000 cells ml⁻¹ with flow rate of 10 ml⁻¹ min⁻¹. Control growth rates, using operculum diameter, were 14 ± 8 μm day⁻¹ and 6 ± 2 μm day⁻¹ for S. balanoides and E. modestus, respectively. Subtle, but significant decreases in E. modestus growth rate were observed in high CO₂ but there were no impacts on shell calcium content and survival by either elevated temperature or CO₂. S. balanoides exhibited no clear alterations in growth rate but did show a large reduction in shell calcium content and survival under elevated temperature and CO₂. These results suggest that a decrease by 0.4 pH_(NBS) units alone would not be sufficient to directly impact the survival of barnacles during the first month post-settlement. However, in conjunction with a 4–5°C increase in temperature, it appears that significant changes to the biology of these organisms will ensue.     

Disappearing plants: why they hide and how they return

Prolonged dormancy is a life-history stage in which mature plants fail to resprout for one or more growing seasons and instead remain alive belowground. Prolonged dormancy is relatively common, but the proximate causes and consequences of this intriguing strategy have remained elusive. In this study we tested whether stored resources are associated with remaining belowground, and investigated the resource costs of remaining belowground during the growing season. We measured stored resources at the beginning and end of the growing season in Astragalus scaphoides, an herbaceous perennial in southwest Montana, USA. At the beginning of the growing season, dormant plants had lower concentrations of stored mobile carbon (nonstructural carbohydrates, NSC) than did emergent plants. Surprisingly, during the growing season, dormant plants gained as much NSC as photosynthetically active plants, an increase most likely due to remobilization of structural carbon. Thus, low levels of stored NSC were associated with remaining belowground, and remobilization of structural carbon may allow for dormant plants to emerge in later seasons. The dynamics of NSC in relation to dormancy highlights the ability of plants to change their own resource status somewhat independently of resource assimilation, as well as the importance of considering stored resources in understanding plant responses to the environment.     

Field evidence for pervasive indirect effects of fishing on prey foraging behavior

The indirect, ecosystem-level consequences of ocean fishing, and particularly the mechanisms driving them, are poorly understood. Most studies focus on density-mediated trophic cascades, where removal of predators alternately causes increases and decreases in abundances of lower trophic levels. However, cascades could also be driven by where and when prey forage rather than solely by prey abundance. Over a large gradient of fishing intensity in the central Pacific's remote northern Line Islands, including a nearly pristine, baseline coral reef system, we found that changes in predation risk elicit strong behavioral responses in foraging patterns across multiple prey fish species. These responses were observed as a function of both short-term ("acute") risk and longer-term ("chronic") risk, as well as when prey were exposed to model predators to isolate the effect of perceived predation risk from other potentially confounding factors. Compared to numerical prey responses, antipredator behavioral responses such as these can potentially have far greater net impacts (by occurring over entire assemblages) and operate over shorter temporal scales (with potentially instantaneous response times) in transmitting top-down effects. A rich body of literature exists on both the direct effects of human removal of predators from ecosystems and predators' effects on prey behavior. Our results draw together these lines of research and provide the first empirical evidence that large-scale human removal of predators from a natural ecosystem indirectly alters prey behavior. These behavioral changes may, in turn, drive previously unsuspected alterations in reef food webs.     

Soft song is a reliable signal of aggressive intent in song sparrows

Animals frequently use signals to modulate aggressive interactions. Establishing that a signal is aggressive or threatening requires demonstrating that it is more commonly used in agonistic contexts, that it predicts subsequent aggressive behaviors by the sender, and that receivers respond differently to this signal. Like many birds, song sparrows (Melospiza melodia) produce a low-amplitude “soft song” vocalization that has been hypothesized to be an aggressive signal. Soft song meets the first two criteria, but previous research has failed to demonstrate that soft song provokes aggression or that receivers even perceive soft song differently from normal loud song. We used a playback experiment with taxidermic mount presentation to test whether territorial male song sparrows respond differently to loud and soft song playbacks. Subjects reacted more strongly to the soft song playback by approaching the mount more closely, increasing wing wave displays, and increasing the proportion of their own songs that were soft songs, with further trends toward increasing the number of flights and attacks. These results confirm that soft song is a conventional signal of aggression in song sparrows and that increased receiver retaliation maintains its reliability.     

Experimental investigation of false positive errors in auditory species occurrence surveys

False positive errors are a significant component of many ecological data sets, which in combination with false negative errors, can lead to severe biases in conclusions about ecological systems. We present results of a field experiment where observers recorded observations for known combinations of electronically broadcast calling anurans under conditions mimicking field surveys to determine species occurrence. Our objectives were to characterize false positive error probabilities for auditory methods based on a large number of observers, to determine if targeted instruction could be used to reduce false positive error rates, and to establish useful predictors of among-observer and among-species differences in error rates. We recruited 31 observers, ranging in abilities from novice to expert, who recorded detections for 12 species during 180 calling trials (66,960 total observations). All observers made multiple false positive errors, and on average 8.1% of recorded detections in the experiment were false positive errors. Additional instruction had only minor effects on error rates. After instruction, false positive error probabilities decreased by 16% for treatment individuals compared to controls with broad confidence interval overlap of 0 (95% CI:--46 to 30%). This coincided with an increase in false negative errors due to the treatment (26%;--3 to 61%). Differences among observers in false positive and in false negative error rates were best predicted by scores from an online test and a self-assessment of observer ability completed prior to the field experiment. In contrast, years of experience conducting call surveys was a weak predictor of error rates. False positive errors were also more common for species that were played more frequently but were not related to the dominant spectral frequency of the call. Our results corroborate other work that demonstrates false positives are a significant component of species occurrence data collected by auditory methods. Instructing observers to only report detections they are completely certain are correct is not sufficient to eliminate errors. As a result, analytical methods that account for false positive errors will be needed, and independent testing of observer ability is a useful predictor for among-observer variation in observation error rates.     

Phenological tracking enables positive species responses to climate change

Earlier spring phenology observed in many plant species in recent decades provides compelling evidence that species are already responding to the rising global temperatures associated with anthropogenic climate change. There is great variability among species, however, in their phenological sensitivity to temperature. Species that do not phenologically "track" climate change may be at a disadvantage if their growth becomes limited by missed interactions with mutualists, or a shorter growing season relative to earlier-active competitors. Here, we set out to test the hypothesis that phenological sensitivity could be used to predict species performance in a warming climate, by synthesizing results across terrestrial warming experiments. We assembled data for 57 species across 24 studies where flowering or vegetative phenology was matched with a measure of species performance. Performance metrics included biomass, percent cover, number of flowers, or individual growth. We found that species that advanced their phenology with warming also increased their performance, whereas those that did not advance tended to decline in performance with warming. This indicates that species that cannot phenologically "track" climate may be at increased risk with future climate change, and it suggests that phenological monitoring may provide an important tool for setting future conservation priorities.     

Underlying causes of habitat-associated differences in size of age-0 walleye pollock (Theragra chalcogramma) in the Gulf of Alaska

Age-0 walleye pollock (Theragra chalcogramma) caught in September in the Gulf of Alaska display habitat-associated differences in standard length (SL). Age-0 fish collected in the region around Sutwik Island and 375?km farther downstream near the Shumagin Islands most likely originate from the Shelikof Strait spawning aggregation. However, age-0 fish resulting from the same spawning aggregation differ in mean size up to 20?mm between areas by September. We examined the otoliths of the larval and age-0 stages of walleye pollock from these two areas in 2000 and 2001 to determine whether growth rate, hatch date, and/or temperature influenced fish size. Circulation models were used to determine whether transport of larvae from an upstream spawning group into the study areas could have occurred. Mean in situ temperature during sampling periods was not defined as a significant factor in altering growth rates. Overlapping hatch date distributions of the larval and age-0 fish in the Shumagin Island area confirmed that the fish were from the Shelikof Strait spawning group. Comparison of hatch date distributions in the upstream Sutwik Island area revealed larger/older larvae from an upstream spawning group mixed with larvae from the Shelikof Strait spawning group. Our results suggest that the offset of 20?mm SL between the groups of age-0 pollock was the result of a combination of enhanced survivorship of early-hatched larvae in the Sutwik area and the introduction and retention of the progeny of another spawning group originating upstream of Shelikof Strait.     

Seasonal patterns in fatty acids of <Emphasis Type="Italic">Calanus hyperboreus</Emphasis> (Copepoda,Calanoida) from Cumberland Sound,Baffin Island,Nunavut

The marine copepod Calanus hyperboreus accumulates large quantities of lipids and essential fatty acids during summer months in Northern oceans. However, few data exist regarding their winter fatty acid profiles, which could be informative regarding the use of lipids by C. hyperboreus to successfully survive and reproduce during times of ice-cover and limited food. The present study compared fatty acids of C. hyperboreus between summer (August 2007 and 2008) and winter (early April 2008 and 2009) in Cumberland Sound, Canada. Summer samples from both years had significantly higher ∑polyunsaturated fatty acids and unsaturation indices (based on μg fatty acid mg dry tissue⁻¹) than winter samples and separated on a principal component analysis due to higher 18:2n-6, 18:4n-3, and 20:5n-3, consistent with phytoplankton consumption. Winter C. hyperboreus had significantly higher ∑monounsaturated fatty acids (MUFA) versus summer samples and separated on the principal component analysis due to higher proportions of 16:1n-7, 20:1n-9, and 22:1n-9, suggesting they were not actively feeding. Based on the seasonal fatty acid comparison, C. hyperboreus was catabolizing specific fatty acids (e.g. 20:5n-3), conserving others (e.g. 22:6n-3), and maintaining or increasing biosynthesis of certain MUFA (e.g. 18:1n-9) during winter. These findings provide insight into the seasonal strategy of acquisition (summer) and utilization (winter) of specific fatty acids by a key Arctic organism and could become important for monitoring changes in fatty acids associated with decreased ice-cover duration due to climate warming.     

Pollution prevention at ports: clearing the air

Seaports are major hubs of economic activity and of environmental pollution in coastal urban areas. Due to increasing global trade, transport of goods through ports has been steadily increasing and will likely continue to increase in the future. Evaluating air pollution impacts of ports requires consideration of numerous sources, including marine vessels, trucks, locomotives, and off-road equipment used for moving cargo. The air quality impacts of ports are significant, with particularly large emissions of diesel exhaust, particulate matter, and nitrogen oxides. The health effects of these air pollutants to residents of local communities include asthma, other respiratory diseases, cardiovascular disease, lung cancer, and premature mortality. In children, there are links with asthma, bronchitis, missed school days, and emergency room visits. The significance of these environmental health impacts requires aggressive efforts to mitigate the problem. Approaches to mitigation encompass a range of possibilities from currently available, low-cost approaches, to more significant investments for cleaner air. Examples of the former include restrictions on truck idling and the use of low-sulfur diesel fuel; the latter includes shore-side power for docked ships, and alternative fuels. A precautionary approach to port-related air pollution would encourage local production of goods in order to reduce marine traffic, greener design for new terminals, and state-of-the art approaches to emissions-control that have been successfully demonstrated at ports throughout the world.