Using vertebrate environmental DNA from seawater in biomonitoring of marine habitats

Conservation and management of marine biodiversity depends on biomonitoring of marine habitats, but current approaches are resource-intensive and require different approaches for different organisms. Environmental DNA (eDNA) extracted from water samples is an efficient and versatile approach to detecting aquatic animals. In the ocean, eDNA composition reflects local fauna at fine spatial scales, but little is known about the effectiveness of eDNA-based monitoring of marine communities at larger scales. We investigated the potential of eDNA to characterize and distinguish marine communities at large spatial scales by comparing vertebrate species composition among marine habitats in Qatar, the Arabian Gulf (also known as the Persian Gulf), based on eDNA metabarcoding of seawater samples. We conducted species accumulation analyses to estimate how much of the vertebrate diversity we detected. We obtained eDNA sequences from a diverse assemblage of marine vertebrates, spanning 191 taxa in 73 families. These included rare and endangered species and covered 36% of the bony fish genera previously recorded in the Gulf. Sites of similar habitat type were also similar in eDNA composition. The species accumulation analyses showed that the number of sample replicates was insufficient for some sampling sites but suggested that a few hundred eDNA samples could potentially capture >90% of the marine vertebrate diversity in the study area. Our results confirm that seawater samples contain habitat-characteristic molecular signatures and that eDNA monitoring can efficiently cover vertebrate diversity at scales relevant to national and regional conservation and management.     

Avoiding the pitfalls of adaptive management implementation in Swedish silviculture

There is a growing demand for alternatives to Sweden’s current dominant silvicultural system, driven by a desire to raise biomass production, meet environmental goals and mitigate climate change. However, moving towards diversified forest management that deviates from well established silvicultural practices carries many uncertainties and risks. Adaptive management is often suggested as an effective means of managing in the context of such complexities. Yet there has been scepticism over its appropriateness in cases characterised by large spatial extents, extended temporal scales and complex land ownership—characteristics typical of Swedish forestry. Drawing on published research, including a new paradigm for adaptive management, we indicate how common pitfalls can be avoided during implementation. We indicate the investment, infrastructure, and considerations necessary to benefit from adaptive management. In doing so, we show how this approach could offer a pragmatic operational model for managing the uncertainties, risks and obstacles associated with new silvicultural systems and the challenges facing Swedish forestry.     

Deriving heart rate variability indices from cardiac monitoring—An indicator of driver sleepiness

Objective: Driver fatigue is considered to be a major contributor to road traffic crashes. Cardiac monitoring and heart rate variability (HRV) analysis is a candidate method for early and accurate detection of driver sleepiness. This study has 2 objectives: to evaluate the (1) suitability of different preprocessing strategies for detecting and removing outlier heartbeats and spectral transformation of HRV signals and their impact of driver sleepiness assessment and (2) relation between common HRV indices and subjective sleepiness reported by a large number of drivers in real driving situations, for the first time.

Methods: The study analyzed >3,500 5-min driving epochs from 76 drivers on a public motorway in Sweden. The electrocardiograph (ECG) data were recorded in 3 studies designed to evaluate the physiological differences between awake and sleepy drivers. The drivers reported their perceived level of sleepiness according to the Karolinska Sleepiness Scale (KSS) every 5?min. Two standard methods were used for identifying outlier heartbeats: (1) percentage change (PC), where outliers were defined as interbeat intervals deviating >30% from the mean of the four previous intervals and (2) standard deviation (SD), where outliers were defined as interbeat interval deviating >4 SD from the mean interval duration in the current epoch. Three standard methods were used for spectral transformation, which is needed for deriving HRV indices in the frequency domain: (1) Fourier transform; (2) autoregressive model; and (3) Lomb-Scargle periodogram. Different preprocessing strategies were compared regarding their impact on derivation of common HRV indices and their relation to KSS data distribution, using box plots and statistical tests such as analysis of variance (ANOVA) and Student’s t test.

Results: The ability of HRV indices to discriminate between alert and sleepy drivers does not differ significantly depending on which outlier detection and spectral transformation methods are used. As expected, with increasing sleepiness, the heart rate decreased, whereas heart rate variability overall increased. Furthermore, HRV parameters representing the parasympathetic branch of the autonomous nervous system increased. An unexpected finding was that parameters representing the sympathetic branch of the autonomous nervous system also increased with increasing KSS level. We hypothesize that this increment was due to stress induced by trying to avoid an incident, because the drivers were in real driving situations.

Conclusions: The association of HRV indices to KSS did not depend on the preprocessing strategy. No preprocessing method showed superiority for HRV association to driver sleepiness. This was also true for combinations of methods for frequency domain HRV indices. The results prove clear relationships between HRV indices and perceived sleepiness. Thus, HRV analysis shows promise for driver sleepiness detection.     

Fungi and wind strongly influence the temporal availability of logs in an old-growth spruce forest.

Coarse woody debris (CWD) is a key habitat for many species in forest ecosystems. To ensure the long-term survival of such species, forest management regimes must include measures that promote dead wood dynamics similar to those of natural forests. Thus, information on CWD dynamics under natural conditions is required, including data pertaining to the underlying agents of disturbance. This study examines modes of mortality, decay rates, and temporal patterns in the availability of Picea abies logs in a Swedish old-growth forest affected by internal, small-scale disturbance. All 684 logs in a 6.6-ha plot were mapped and classified into one of six decay classes. Logs in the early stages of decay were examined for the presence of heart-rot fungi. Six years later all logs were re-inventoried, including newly formed logs. Matrix models based on the transition rates between decay classes showed that it took about 60 years for 90% of the logs to decay beyond class 6 (a deformed trunk with soft wood). Large logs (> 26 cm) decayed 40% more slowly than small logs (< 25 cm). The initial volume of logs was 37.6 m3/ha but increased to 44.8 m3/ha after six years. In addition, there was a large shift in the decay-class distribution. The volume of logs in early and late decay classes increased by 71% and 45%, respectively, while the volume of logs in the intermediate decay classes decreased by 32%. The fluctuations appear to result from pulses in mortality, driven by a combination of strong winds and the heart-rot fungus, Phellinus chrysoloma, which was present in more than 30% of all logs at an early stage of decay. These results show that large temporal fluctuations in dead wood also occur in the absence of large-scale disturbance, and that heart-rot fungi are important factors driving the overall dynamics of dead wood. Since many wood-inhabiting species are naturally rare and have very specific substrate demands, such temporal variability in dead wood availability may have effects on biodiversity and should be taken into account when designing small, protected forest areas.     

Hyperbenthic crustacean fauna of the Gullmarfjord area (western Sweden): Species richness,seasonal variation and long-term changes

Distribution of 118 species of Amphipoda, Mysidacea and Decapoda, sampled in 1984 and 1985 with an epibenthic sledge along a transect from the Skagerrak to the inner Gullmarfjord (Sweden), is described. Amphipods were richest in species in the Skagerrak, while most mysid species were taken at the sill. Deca-impoverished and the Lilljeborgidae, earlier represented by three species, had disappeared since the 1930s. The changes are most likely caused by eutrophication of the fjord. and deep-basin samples; (III) sill samples; (IV) Skagerrak samples. Characteristic species of the groups were identified with a pseudoF-test. Detrended Correspondence Analysis indicated seasonal faunal changes at 42 and 72 m depths. At both depths a shift in numerical dominance from amphipods to mysids was found in winter. The most pronounced changes seemed to occur at 42 m. Changes in composition and abundance of species during a renewal of the bottom water indicated that mysids were influenced by presumed near-bottom currents, while amphipods and decapods were not. Comparison of the present amphipod fauna with the fauna in 1933–37 revealed significant differences. The fauna in the deep basin was impoverished and the Lilljeborgidae, earlier represented by three species, had disappeared since the 1930s. The changes are most likely caused by eutrophication of the fjord.     

Gill pump characteristics of the soft clamMya arenaria

Pumping rates in the soft clamMya arenaria, collected in June 1987 in the Great Belt, Denmark, were determined as rates at which clams cleared suspensions of algae,Dunaliella marina. The frictional resistance of the siphons to water flow was estimated by studying the effect of excision of the siphons at the base on the rate of water pumping. The frictional resistance was also calculated from the Poiseuille equation. Excision of the siphons had no measurable effect on the pumping rate, and calculations indicated pressure losses ranging between 0.3 to 1.2 mm H₂O. This is consistent with the conception that the capacity of filter feeding bivalves to process large amounts of water depends upon a low resistance to water flow in the siphons.     

Particle capture in the musselMytilus edulis: The role of latero-frontal cirri

Microscope video graphs of particle paths near one-filament-thick mussel gill preparations, stimulated with a nerve transmitter (10^–6 M serotonin which restores normal ciliary activity), were used to disclose the capture of 6 m algal cells. Suspended algal cells carried with the water were stopped for a while at the entrance to the interfilament gap by the action of the latero-frontal cirri (Ifc), and transferred to the frontal side of the filament to be transported towards the marginal food groove. The event of transfer took place during approximately a time interval of 1150 to 1/25 s. To gain a better understanding of the capture mechanism and retention efficiency versus particle size, the flow through and around the Ifc was theoretically estimated. Normally beating Ifc create periodic, unsteady, three-dimensional flows at the entrance to the interfilament canal. During the active beat most of the water is deflected to flow around the branching cilia of the Ifc while some of the water is strained by these. Large particles (> 4 m) are stopped and transferred to the frontal current, whereas smaller particles either follow the flow around the Ifc and escape or they are stopped by the branching cilia.     

Concentrations of inorganic elements in bottled waters on the Swedish market

This study presents the concentrations of about 50 metals and ions in 33 different brands of bottled waters on the Swedish market. Ten of the brands showed calcium (Ca) concentrations ≤10 mg L⁻¹ and magnesium (Mg) levels <3 mg L⁻¹, implying very soft waters. Three of these waters had in addition low concentrations of sodium (Na; <7 mg L⁻¹), potassium (K; <3 mg L⁻¹) and bicarbonate (HCO₃ ≤31 mg L⁻¹). These brands were collected from barren districts. Nine of the brands were collected from limestone regions. They showed increased Ca-levels exceeding 50 mg L⁻¹ with a maximum of 289 mg L⁻¹. Corresponding Mg-levels were also raised in two brands exceeding 90 mg L⁻¹. Two soft and carbonated waters were supplemented with Na₂CO₃ and NaCl, resulting in high concentrations of Na (644 and 648 mg L⁻¹) and chloride (Cl; 204 and 219 mg L⁻¹). Such waters may make a substantial contribution to the daily intake of NaCl in high water consumers. The storage of carbonated drinking water in aluminum (Al) cans increased the Al-concentration to about 70 μg L⁻¹. Conclusion As there was a large variation in the material as regards concentrations of macro-elements such as Ca, Mg, Na, K and Cl. Supplementation with salts, e.g., Na₂CO₃, K₂ CO₃ and NaCl, can lead to increased concentrations of Na, K and Cl, as well as decreased ratios of Ca/Na and larger ratios of Na/K. Water with high concentrations of e.g., Ca and Mg, may make a substantial contribution to the daily intake of these elements in high water consumers. Al cans are less suited for storage of carbonated waters, as the lowered pH-values may dissolve Al. The levels of potentially toxic metals in the studied brands were generally low.     

High frequency of extra-pair paternity in Swedish pied flycatchers revealed by allozyme electrophoresis and DNA fingerprinting

Summary We used two genetic techniques to study multiple parentage in the pied flycatcher (Ficedula hypoleuca). Allozyme electrophoresis of 131 nestlings from 22 clutches sampled in 1982 and 1987 showed that one nestling had a mismatched allele compared with its putative parents. In one additional nest the devitation from Mendelian inheritance of parental genotypes suggested extra-pair paternity. The calculated probability of detecting multiple paternity from the genetic variation at four loci was 0.115. The estimated population frequency of extra-pair fertilization (EPF) was 13%, based on two mismatches and the probability of detecting multiple paternity. The seven families (n = 38 nestlings) in the 1987 sample were also analyzed by DNA fingerprinting using the M13 and Jeffrey's 33.15 probes. Overall, 24% (n = 9) of the nestlings analyzed were genetically inconsistent with their putative father, with EPF occurring in three (43%) of the seven clutches. One nestling originated by intraspecific brood parasitism. Comparison of the two techniques using the same samples showed that allozyme electrophoresis has a resolution only 11% of that of DNA fingerprinting, close to the calculated probability of detecting multiple paternity (0.115). Both techniques suggest that extra-pair fertilization is relatively common in the Swedish populations investigated compared to the low frequency reported from a Norwegian population. Correspondence to: H.P. Gelter at the present address