Acoustic detection and acoustic habitat characterisation of the critically endangered white-bellied heron (Ardea insignis) in Bhutan

1. Growing developmental activities, such as hydropower construction, farm roads, and other human activities, are affecting the critically endangered white-bellied heron (WBH). Out of a known global population of 60, 28 individuals inhabit the river basin area and freshwater lakes and ponds of Bhutan. Several constraints impede continuous monitoring of endangered species, such as the isolated and cryptic nature of the species and the remoteness of its habitat; to date, there are no long-term reference data or techniques implemented for continuous monitoring of this species.
2. In this study, we designed acoustic detection and habitat characterisation methods using long-duration recordings from three habitat areas in Bhutan. Acoustic indices were extracted and used to implement a species-specific call detector and to generate habitat soundscape representations. Using WBH calls annotated in month-long recordings from a known site, a novel indices-based detector was implemented and tested. A total of 960 hr of continuous audio recordings from three habitats in Bhutan were analysed.
3. We found that a species call detector implemented using a combination of acoustic indices (that includes measures of spectral and temporal entropy and different angles of spectral ridges) has a correct detection rate of 81%. Additionally, visual inspection of the species’ acoustic habitat using long-duration false-colour spectrograms enabled qualitative assessment of acoustic habitat structure and other dominant acoustic events.
4. This study proposes a combined approach of species acoustic detection and habitat soundscape analysis for holistic acoustic monitoring of endangered species. As a direct outcome of this work, we documented acoustic reference data on the critically endangered WBH from multiple habitat areas and have analysed its temporal vocalisation patterns across sites.

     

Diurnal variation in freshwater ecoacoustics: Implications for site-level sampling design

1. Ecoacoustic methods are increasingly used to monitor the state of populations and ecosystems. In freshwater environments, they present the clear advantages of being non-invasive, reducing bias, and providing continuous observations instead of only limited sampling snapshots in time. However, similar to standard bioassessment methods, temporal variation and choice of indicators can greatly influence ecoacoustic assessments, highlighting the importance of sampling and analysis design.
2. In this study, we quantified diurnal variation in underwater sound and its effect on sampling regimes for two waterholes in the Einasleigh River, Northern Australia. Recording continuously for 6 days, and subsampling 5 s every 10 min, we found 22 distinct sounds that were emitted by fish, Hemiptera and Coleoptera as well as another 22 of abiotic or unknown origin.
3. Through rarefaction analyses, we found that subsampling the data to 60% of the recorded sound events resulted in capture of most of the 44 identified sound types. Temporal heterogeneity—patchy sound events through time—needs to be considered when maximising detected sound events. Reducing the sampling interval from every 10 min to half-hourly or hourly had a much greater effect on capturing all sound types compared to the number of days recorded or the length of the recording. Overall, only 10–20% of the sound events need to be annotated for most sound types to be described; for example, restricting analysis of the days recorded to only three and the recording interval to 0.5–1 s. Acoustic indices were dominated by three main event types—a diurnally flowing creek, a nocturnal chorus of Hemiptera, as well as a dawn chorus of terapontid fishes.
4. We conclude with two key messages: First, a select group of informative signals can be monitored using very simple methods—namely, converting an audio stream into indices using freely available software. Second, however, to detect less acoustically dominant sound events, manual annotation or single call processing will still be needed. While these findings are encouraging, similar analysis will need to be conducted within other freshwater ecosystems before general conclusions about optimal sampling regimes can be drawn.

     

Six steps towards operationalising freshwater ecoacoustic monitoring

1. Applications in bioacoustics and its sister discipline ecoacoustics have increased exponentially over the last decade. However, despite knowledge about aquatic bioacoustics dating back to the times of Aristotle and a vast amount of background literature to draw upon, freshwater applications of ecoacoustics have been lagging to date.
2. In this special issue, we present nine studies that deal with underwater acoustics, plus three acoustic studies on water-dependent birds and frogs. Topics include automatic detection of freshwater organisms by their calls, quantifying habitat change by analysing entire soundscapes, and detecting change in behaviour when organisms are exposed to noise.
3. We identify six major challenges and review progress through this special issue. Challenges include characterisation of sounds, accessibility of archived sounds as well as improving automated analysis methods. Study design considerations include characterisation analysis challenges of spatial and temporal variation. The final key challenge is the so far largely understudied link between ecological condition and underwater sound.
4. We hope that this special issue will raise awareness about underwater soundscapes as a survey tool. With a diverse array of field and analysis tools, this issue can act as a manual for future monitoring applications that will hopefully foster further advances in the field.

     

The Application of the Acoustic Complexity Indices (ACI) to Ecoacoustic Event Detection and Identification (EEDI) Modeling

In programs of acoustic survey, the amount of data collected and the lack of automatic routines for their classification and interpretation can represent a serious obstacle to achieving quick results. To overcome these obstacles, we are proposing an ecosemiotic model of data mining, ecoacoustic event detection and identification (EEDI), that uses a combination of the acoustic complexity indices (ACIt_f, ACIf_t, and ACIf_te) and automatically extracts the ecoacoustic events of interest from the sound files. These events may be indicators of environmental functioning at the scale of individual vocal species (e.g., behavior, phenology, and dynamics), the acoustic community (e.g., dawn and dusk chorus), the sound marks (e.g., the flag species of a community), or the soundscape (e.g., sonotope types). The EEDI model is represented by three procedural steps: 1) selecting acoustic data according to environmental variables, 2) detecting the events by creating an ecoacoustic event space (EES) produced by plotting ACIf_t and its evenness (ACIf_te), 3) identifying events according to the level of correlation between the acoustic signature (ACIt_f) of the detected events and an ad hoc library of previously identified events. The EEDI procedure can be extensively used in basic and applied research. In particular, EEDI may be used in long-term monitoring programs to assess the effect of climate change on individual vocal species behavior (fishes, frogs, birds, mammals, and arthropods), population, and acoustic community dynamics. The EEDI model can be also used to investigate acoustic human intrusion in natural systems and the effect in urban areas.     

Acoustic monitoring reveals diversity and surprising dynamics in tropical freshwater soundscapes

1. Freshwater systems are globally threatened and in need of enhanced monitoring and assessment. We applied soundscape recording and analysis—which presents an opportunity for long-term, high-resolution animal community monitoring and assessment—to a freshwater context to better understand the acoustic diversity and dynamics of these systems.
2. We recorded the aquatic soundscape of a Neotropical freshwater swamp in Costa Rica for 23 days in January and February 2015 during the dry season. We classified biological sound types in these recordings and developed measurements of richness and occupancy based on this classification. We also calculated six complementary acoustic indices to assess soundscape diversity and daily and longer-term soundscape dynamics, and we examined correlations between these acoustic indices and sound type metrics.
3. We found rich soundscapes in which biological sounds were almost always present, and we classified 18 sound types that we attribute to aquatic insects. These sound types showed distinct daily patterns and exhibited temporal and spectral acoustic niche partitioning. Sound type richness was most correlated with the number of peaks index (correlation = .36; p < .001), while sound type occupancy was most correlated with the Bioacoustic Index (correlation = .92; p < .001). In contrast to generally high levels of acoustic activity, there were brief (approximately 1 hr), unexpected quiet periods around dawn and dusk.
4. This study represents an early attempt to comprehensively describe tropical freshwater soundscapes in a systematic and quantitative manner. We demonstrate that sound type classification and the quantification of acoustic occupancy capture aspects of soundscape diversity and dynamics that are complementary to those assessed by acoustic indices. Our analyses reveal that the soundscapes of this tropical wetland were diverse and exhibited daily dynamics that differed from those found in other ecosystems.

     

Looking for love under the ice: Using passive acoustics to detect burbot (Lota lota: Gadidae) spawning activity

1. Burbot (Lota lota: Gadidae) is a difficult species to manage effectively due to its preference for deep-water habitats and under-ice spawning behaviour, resulting in a poor understanding of its reproductive activity. However, the use of acoustic signalling by burbot as part of their mating system has recently been described and this behaviour may provide a means of investigating questions regarding the spatial and temporal distribution of spawning aggregations using passive acoustic monitoring.
2. We used audio and video recording to confirm that burbot vocalise and that these vocalisations can be detected under field conditions as well as to characterise the relationship between burbot acoustic signalling and spawning behaviour. We also evaluated the feasibility of locating and monitoring burbot spawning aggregations in real time using passive acoustics.
3. Burbot vocalisations were difficult to identify with only about 6% of the recordings containing calls being successfully identified as such in the field. Burbot vocalised more often between sundown and sunrise than during daylight hours. Calls recorded at night tended to be lower frequency, longer duration, and have lower bandwidth than those made during the day.
4. Burbot vocalisations could not be recorded in conjunction with video recordings of spawning activity, indicating that burbot may not call during active spawning, but may use acoustic communication to signal the onset of reproductive readiness and to form pre-spawning aggregations.
5. While burbot calls were readily identifiable, observers had a difficult time identifying burbot calls in real time under field conditions. Passive acoustic monitoring demonstrates considerable potential as a management tool to locate burbot spawning grounds and identify periods of activity, but may not be an appropriate technique for monitoring spawning activity in real time.

     

Assessing the value of acoustic indices to distinguish species and quantify activity: A case study using frogs

1. Soundscapes can provide information about a wide range of habitats and species through the recording of vocalisations over long temporal scales. Because of the large volumes of data collected, computational approaches, such as the application of acoustic indices, are required to extract useful information from long-duration recordings.
2. Acoustic indices summarise various soundscape features into frequency ranges over defined time intervals and can aid in the visual exploration, detection, and analysis of species vocalisation patterns. Here, we examine the performance of combinations of three acoustic indices commonly used in visual exploration, the acoustic complexity index, the temporal entropy spectrum index, and the event spectrum index, and assess their ability to distinguish species and describe acoustic features commonly used to detect species and analyse activity. Our case study focuses on three frog species with distinct call structures from Bickerton Island, Northern Territory, Australia. Call structure was categorised based on the number of pulses and harmonics.
3. We summarised acoustic activity by calculating acoustic indices in 256 equal-sized bins over the entire the frequency spectrum, for 30-s intervals, and found that acoustic index values could be used to distinguish species and describe acoustic features. The acoustic complexity index was the most effective index for distinguishing species. To describe acoustic features, we examined correlations between acoustic index values and summarised acoustic features, including call rate, total duration, loudness and signal-to-noise ratio. In single-pulsed species with no harmonics, we found spectral index values were significantly and sometimes strongly correlated with acoustic features. In comparison, species with harmonics were found to be weakly and less frequently correlated with acoustic features even if sampled calls were loud and have high signal-to-noise ratio. We suggest that acoustic indices have the potential to describe acoustic features in single-pulsed species but are limited in those with harmonics.
4. We conclude that acoustic indices can be a useful tool to distinguish some anuran species and to broadly understand specific acoustic features used to analyse calling activity over long periods of time.
5. Further research is required to better understand the relationships between acoustic indices and acoustic features to determine the general utility of indices to detect and distinguish audible species and to identify other acoustic features of various taxa.

     

Automated acoustic monitoring of endangered common spadefoot toad populations reveals patterns of vocal activity

1. In the context of global amphibian decline, monitoring and restoration programmes are important. Acoustic monitoring is a possible approach for underwater vocalising species like the rapidly declining European common spadefoot toad (Pelobates fuscus). In this study, our aim was to design a dedicated software detector to be used in combination with programmable audio recorders to process the large amount of data generated by long-term acoustic monitoring and to use it for investigating the seasonal and circadian patterns of P. fuscus vocal activity.
2. The software detector targets advertisement calls of the species. Based on acoustic analysis of that call, we developed a detector that utilises both frequency and time features of the calls. Data collected during three breeding seasons in four known or potential P. fuscus breeding sites of north-eastern France were used to build a ground truth in order to test the performance of the detector. Then, we used the detector for analysing four acoustic monitoring campaigns conducted in two different sites over two breeding seasons to gain insight into the seasonal and circadian patterns of vocal activity of this species.
3. Evaluation of the P. fuscus call detector against a ground truth returned false-positive rates below 1.5% and true-positive rates ranging from 53% to 73%. These figures are compatible with long-term monitoring of the presence of the species. Running the software detector on standard hardware, the computation time for post-processing the 360 hr of a typical 3-month monitoring campaign was less than 1 day.
4. The seasonal pattern of P. fuscus underwater vocal activity is more complex than previously recognised. Over the whole ostensible 3-month breeding season, the actual time window for vocalising and breeding can last from a few days up to several weeks and may be split into clearly distinct episodes. When vocalisations occurred at both night- and daytime, the circadian vocal activity of P. fuscus occasionally proceeded uninterrupted for 24 hr but usually a several hour lull occurred immediately prior to sunset. When vocalisations occurred at both night- and daytime, the vocal activity pattern followed a bimodal distribution with a nocturnal highest peak of activity and a second peak occurring in the morning.
5. Our results demonstrate that it is feasible to monitor presence of P. fuscus in north-eastern France using a dedicated software detector combined with programmable audio recorders. Based on the outcomes of the detector applied to long-term audio data sets, we reveal temporal patterns of the vocal activity of the species and subsequently provide recommendations for attended and unattended acoustic monitoring.

     

Acoustic activity across a seabird colony reflects patterns of within-colony flight rather than nest density

Passive acoustic monitoring is increasingly being used as a cost-effective way to study wildlife populations, especially those that are difficult to census using conventional methods. Burrow-nesting seabirds are among the most threatened birds globally, but they are also one of the most challenging taxa to census, making them prime candidates for research into such automated monitoring platforms. Passive acoustic monitoring has the potential to determine presence/absence or quantify burrow-nesting populations, but its effectiveness remains unclear. We compared passive acoustic monitoring, tape-playbacks and GPS tracking data to investigate the ability of passive acoustic monitoring to capture unbiased estimates of within-colony variation in nest density for the Manx Shearwater Puffinus puffinus. Variation in acoustic activity across 12 study plots on an island colony was examined in relation to burrow density and environmental factors across 2 years. As predicted fewer calls were recorded when wind speed was high, and on moon-lit nights, but there was no correlation between acoustic activity and the density of breeding birds within the plots as determined by tape-playback surveys. Instead, acoustic indices correlated positively with spatial variation in the in-colony flight activity of breeding individuals detected by GPS. Although passive acoustic monitoring has enormous potential in avian conservation, our results highlight the importance of understanding behaviour when using passive acoustic monitoring to estimate density and distribution.     

Pond Acoustic Sampling Scheme: A draft protocol for rapid acoustic data collection in small waterbodies

1. Freshwater conservation is vital to the maintenance of global biodiversity. Ponds are a critical, yet often under‐recognized, part of this, contributing to overall ecosystem functioning and diversity. They provide habitats for a range of aquatic, terrestrial, and amphibious life, often including rare and declining species.
2. Effective, rapid, and accessible survey methods are needed to enable evidence‐based conservation action, but freshwater taxa are often viewed as “difficult”—and few specialist surveyors are available. Datasets on ponds are therefore limited in their spatiotemporal coverage.
3. With the advent of new recording technologies, acoustic survey methods are becoming increasingly available to researchers, citizen scientists, and conservation practitioners. They can be an effective and noninvasive approach for gathering data on target species, assemblages, and environmental variables. However, freshwater applications are lagging behind those in terrestrial and marine spheres, and as an emergent method, research studies have employed a multitude of different sampling protocols.
4. We propose the Pond Acoustic Sampling Scheme (PASS), a simple protocol to allow a standardized minimal sample to be collected rapidly from small waterbodies, alongside environmental and methodological metadata. This sampling scheme can be incorporated into a variety of survey designs and is intended to allow access to a wide range of participants, without requiring complicated or prohibitively expensive equipment.
5. Adoption of this sampling protocol would enable consistent sound recordings to be gathered by researchers and conservation organizations, and allow the development of landscape‐scale surveys, data sharing, and collaboration within an expanding freshwater ecoacoustic community—rather than individual approaches that produce incompatible datasets. The compilation of standardized data would improve the prospects for effective research into the soundscapes of small waterbodies and aid freshwater conservation efforts.

     

Spatio-temporal heterogeneity in river sounds: Disentangling micro- and macro-variation in a chain of waterholes

1. Passive acoustic monitoring is gaining momentum as a viable alternative method to surveying freshwater ecosystems. As part of an emerging field, the spatio-temporal replication levels of these sampling methods need to be standardised. However, in shallow waters, acoustic spatio-temporal patchiness remains virtually unexplored.
2. In this paper, we specifically investigate the spatial heterogeneity in underwater sounds observed within and between waterholes of an ephemeral river at different times of the day and how it could affect sampling in passive acoustic monitoring.
3. We recorded in the Einasleigh River, Queensland in August 2016, using a linear transect of hydrophones mounted on frames. We recorded four times a day: at dawn, midday, dusk, and midnight. To measure different temporal and spectral attributes of the recorded sound, we investigated the mean frequency spectrum and computed acoustic indices.
4. Both mean frequency spectrum and index analyses revealed that the site and diel activity patterns significantly influenced the sounds recorded, even for adjacent sites with similar characteristics along a single river. We found that most of the variation was due to temporal patterns, followed by between-site differences, while within-site differences had limited influence.
5. This study demonstrates high spatio-temporal acoustic variability in freshwater environments, linked to different species or species groups. Decisions about sampling design are vital to obtain adequate representation. This study thus emphasises the need to tailor spatio-temporal settings of a sampling design to the aim of the study, the species and the habitat.

     

Singing streams: Describing freshwater soundscapes with the help of acoustic indices

1. Understanding soundscapes, that is, the totality of sounds within a location, helps to assess nature in a more holistic way, providing a novel approach to investigating ecosystems. To date, very few studies have investigated freshwater soundscapes in their entirety and none across a broad spatial scale.
2. In this study, we recorded 12 freshwater streams in South East Queensland continuously for three days and calculated three acoustic indices for each minute in each stream. We then used principal component analysis of summary statistics for all three acoustic indices to investigate acoustic properties of each stream and spatial variation in their soundscapes.
3. All streams had a unique soundscape with most exhibiting diurnal variation in acoustic patterns. Across these sites, we identified five distinct groups with similar acoustic characteristics. We found that we could use summary statistics of AIs to describe daytimes across streams as well. Most difference in stream soundscapes was observed during the daytime with significant variation in soundscapes both between hours and among sites.
4. Synthesis and Application. We demonstrate how to characterize stream soundscapes by using simple summary statistics of complex acoustic indices. This technique allows simple and rapid investigation of streams with similar acoustic properties and the capacity to characterize them in a holistic and universal way. While we developed this technique for freshwater streams, it is also applicable to terrestrial and marine soundscapes.

     

Monitoring the acoustic activity of an aquatic insect population in relation to temperature,vegetation and noise

1. Acoustic population monitoring is a noninvasive method that can be deployed continuously over long periods of time and at large spatial scales. One of the newly discovered threats acting on biological diversity is anthropogenic noise. High levels of anthropogenic noise occur in aquatic environments, yet their effects on animals living in freshwater habitats have very rarely been investigated.
2. Here, we used acoustic monitoring and automatic detection to assess the acoustic activity of a population of a soniferous freshwater insect.
3. The sounds emitted by the corixid Micronecta scholtzi were recorded in a Mediterranean pond with an array of 12 hydrophones. An automatic analysis based on a measure of the amplitude found in the frequency band of M. scholtzi was developed to assess the level of acoustic activity. We used functional linear models, accounting for the periodicity of the calling behaviour, to estimate the possible effect of temperature, vegetation and a noise due to an immersed engine.
4. The automatic analysis was validated as an efficient method to measure the acoustic activity. The monitoring revealed a clear 24-hr pattern in the acoustic activity of M. scholtzi and three peaks of activity during the morning. Functional linear models revealed negative effects of both temperature and vegetation and showed that an engine noise, played back for 2 hr during the night, elicited an increase in the level of acoustic activity of the population. Moreover, a cross-correlation procedure showed that noise delayed the acoustic activity of the population.
5. Our results suggest that acoustic survey and automatic detection are efficient methods to monitor the acoustic activity of an insect population especially in response to an anthropogenic disturbance.

     

Changes in functional composition and diversity of waterbirds: The roles of water level and submerged macrophytes

1. Water level and submerged macrophytes are critical players for the functioning of shallow lake ecosystems; understanding how waterbird communities respond to changes in both can have important implications for conservation and management. Here, we evaluated the effects of changes in water level and submerged macrophyte status on wintering waterbird community size, functional group abundances, functional diversity (FD), and community assembly by using a dataset compiled over 50 years.
2. We built generalised linear models to evaluate the effects of water level and submerged macrophyte status on the above-listed attributes of the waterbird communities by using mid-winter waterbird censuses, water level measurements, and submerged macrophyte surveys, along with submerged macrophyte macrofossil records from two shallow lakes in Turkey. Using a relevant set of functional traits, we defined functional groups, calculated four FD measures, and simulated null distributions of the FD measures for assessing assembly rules.
3. We found that macrophyte-dominated years had significantly higher abundances of waterbirds in one of the study lakes, and had more diving herbivores and omnivores in both lakes, while diving/scooping fish-eating waterbird abundance was lower in macrophyte-dominated years. Community size in Lake Beyşehir exhibited a negative association with water level; surprisingly, however, none of the functional group abundances and FD indices were significantly related to water level.
4. In our study communities, standardised effect sizes of functional richness and functional dispersion—two indices that are particularly sensitive to community assembly processes—were mostly lower than those of randomly assembled communities, which implies functional clustering. Shifts to a scarce-macrophyte state were associated with increases in these two indices, possibly due to either changes in the relative strength of environmental filtering and limiting similarity in community assembly or sampling of transitional communities. Further studies covering a wider range of the trophic/macrophyte status spectrum are needed to be certain.
5. The results of this study indicate that shifts between abundant and scarce-macrophyte states can have significant effects on wintering waterbird abundances, FD and community assembly. The results also suggest that shallow lakes in macrophyte-dominated states can support more wintering waterbirds, especially diving omnivores, some of which are globally threatened.

     

Large-scale bioacoustic monitoring to elucidate the distribution of a non-native katydid

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Decoding the ‘zeep’ complex: quantitative analysis of interspecific variation in the nocturnal flight calls of nine wood warbler species (Parulidae spp.)

ABSTRACT

The “zeep” complex consists of nine birds that produce nocturnal flight calls with similar acoustic features. Our inability to distinguish these calls inhibits the acoustic monitoring of these species. We test the hypothesis that flight calls of nine warblers in the “zeep” complex show sufficient acoustic differences to allow differentiation. We investigate divergence in these vocalizations by recording birds held for banding and collecting additional recordings from sound libraries. We used three approaches to compare calls between species: analysis of variance in acoustic properties, discriminant analysis of acoustic properties, and spectrographic cross-correlation. The first approach revealed five species that were different in one or more acoustic properties. The second approach revealed a level of assignment to the correct species (73%) that exceeded levels expected by chance (36%). The third approach revealed calls of seven species to be significantly more similar to conspecific calls than heterospecific calls. Our results suggest the calls of many members of the “zeep” complex exhibit species-specific differences in structure, which may allow differentiation of at least five “zeep” species based on call alone. We advocate for the combined use of these three approaches for the comparison of “zeep” calls in future flight call studies.     

Detecting invertebrate ecosystem service providers in orchards: traditional methods versus barcoding of environmental DNA in soil

1. The objective of this study was to assess barcoding of environmental DNA as a method for monitoring invertebrate ecosystem service providers in soil samples.
2. We selected 26 invertebrate ecosystem service providers that occur in New Zealand kiwifruit or apple orchards and produced mitochondrial cytochrome c oxidase gene subunit I (cytochrome oxidase I) and/or 28S ribosomal DNA sequences for each. Specific barcode primers were designed for each invertebrate ecosystem service provider and tested, along with generic barcoding cytochrome oxidase I primers, for their ability to detect DNA from invertebrate ecosystem service providers that had been added to sterilized and unsterilized soil samples.
3. Although the specific primers accurately detected the invertebrate ecosystem service providers in more than 96% of the samples, the generic cytochrome oxidase I primers detected only 37% of the invertebrate ecosystem service providers added to the sterilized samples and 2.5% in the unsterilized samples.
4. In a field test, we compared metabarcoding with traditional invertebrate trapping methods to detect the invertebrate ecosystem service providers in 10 kiwifruit and 10 apple orchards. All invertebrate ecosystem service providers were collected in traps in at least one orchard, but very few were identified by metabarcoding of soil environmental DNA.
5. Although the specific primers can be used as a tool for monitoring invertebrate ecosystem service providers in soil samples, methodological improvements are needed before metabarcoding of soil environmental DNA can be used to monitor these taxa.

     

Considerations for metabarcoding‐based port biological baseline surveys aimed at marine nonindigenous species monitoring and risk assessments

1. Monitoring introduction and spread of nonindigenous species via maritime transport and performing risk assessments require port biological baseline surveys. Yet, the comprehensiveness of these surveys is often compromised by the large number of habitats present in a port, the seasonal variability, and the time‐consuming morphological approach used for taxonomic identification. Metabarcoding represents a promising alternative for rapid comprehensive port biological baseline surveys, but its application in this context requires further assessments.
2. We applied metabarcoding (based on barcodes of the cytochrome c oxidase subunit I and of the 18S ribosomal RNA gene) to 192 port samples collected (a) from diverse habitats (water column—including environmental DNA and zooplankton, sediment, and fouling structures), (b) at different sites (from inner to outer estuary), and iii) during the four seasons of the year.
3. By comparing the biodiversity metrics derived from each sample group, we show that each sampling method resulted in a distinct community profile and that environmental DNA alone cannot substitute for organismal sampling, and that, although sampling at different seasons and locations resulted in higher observed biodiversity, operational results can be obtained by sampling selected locations and seasons.
4. By assessing the taxonomic composition of the samples, we show that metabarcoding data allowed the detection of previously recorded nonindigenous species as well as to reveal presence of new ones, even if in low abundance.
5. Synthesis and application. Our comprehensive assessment of metabarcoding for port biological baseline surveys sets the basics for cost‐effective, standardized, and comprehensive monitoring of nonindigenous species and for performing risk assessments in ports. This development will contribute to the implementation of the recently entered into force International Convention for the Control and Management of Ships' Ballast Water and Sediments.

     

A community context for aggression? Multi‐species audience effects on territorial aggression in two species of Paridae

1. Territorial aggression in birds is widely observed and is commonly linked to sex, age, body size, physiology, seasonal cues, food resource, urbanization, and a variety of social contexts including conspecific audience effects. However, little is known about the heterospecific audience effects on territorial aggression.
2. Here, we address an emerging idea that heterospecific audience effects may be pervasive influences in the social lives of free‐living birds. We tested the hypothesis that the composition, number, and relative body size of heterospecific audiences observing an aggressive contest will influence the response probability and intensity of aggression displayed.
3. We subjected two Paridae species, tufted titmouse (TUTI, Baeolophus bicolor) and Carolina chickadee (CACH, Poecile carolinensis), to playbacks of aggressive calls during a breeding season in north‐central Florida. At widely spaced playback sites (N = 134) in woodland habitats, we characterized the makeup of heterospecific audiences, aggression type (intra vs. interspecific territoriality), local population density, and various environmental factors (tree density, wind speed, and noise level) that are likely to influence territorial aggression.
4. We found that the presence of heterospecific audiences increased TUTI aggression levels and that both parids were more likely to respond to playback stimuli when their audiences had higher heterospecific diversity (more heterospecific individuals and species). We also found TUTI were more likely to respond when CACH were present but not vice versa.
5. In conclusion, we found evidence that heterospecific audiences significantly influenced the metrics of territorial aggression of free‐living animals and we suggest that the definition of audience effects on the behavior of free‐living animals be expanded to incorporate heterospecific audiences.

     

Resolving the identification of weak-flying insects during flight: a coupling between rigorous data processing and biology

1. Bioacoustic methods play an increasingly important role for the detection of insects in a range of surveillance and monitoring programmes.
2. Weak-flying insects evade detection because they do not yield sufficient audio information to capture wingbeat and harmonic frequencies. These inaudible insects often pose a significant threat to food security as pests of key agricultural crops worldwide.
3. Automatic detection of such insects is crucial to the future of crop protection by providing critical information to assess the risk to a crop and the need for preventative measures.
4. We describe an experimental set-up designed to derive audio recordings from a range of weak-flying aphids and beetles using an LED array.
5. A rigorous data processing pipeline was developed to extract meaningful features, linked to morphological characteristics, from the audio and harmonic series for six aphid and two beetle species.
6. An ensemble of over 50 bioacoustic parameters was used to achieve species discrimination with a success rate of 80%. The inclusion of the dominant and fundamental frequencies improved prediction between beetles and aphids because of large differences in wingbeat frequencies.
7. At the species level, error rates were minimized when harmonic features were supplemented by features indicative of differences in species' flight energies.