Temporally precise population coding of dynamic sounds by auditory cortex.

Fluctuations in the amplitude envelope of complex sounds provide critical cues for hearing, particularly for speech and animal vocalizations. Responses to amplitude modulation (AM) in the ascending auditory pathway have chiefly been described for single neurons. How neural populations might collectively encode and represent information about AM remains poorly characterized, even in primary auditory cortex (A1). We modeled population responses to AM based on data recorded from A1 neurons in awake squirrel monkeys and evaluated how accurately single trial responses to modulation frequencies from 4 to 512 Hz could be decoded as functions of population size, composition, and correlation structure. We found that a population-based decoding model that simulated convergent, equally weighted inputs was highly accurate and remarkably robust to the inclusion of neurons that were individually poor decoders. By contrast, average rate codes based on convergence performed poorly; effective decoding using average rates was only possible when the responses of individual neurons were segregated, as in classical population decoding models using labeled lines. The relative effectiveness of dynamic rate coding in auditory cortex was explained by shared modulation phase preferences among cortical neurons, despite heterogeneity in rate-based modulation frequency tuning. Our results indicate significant population-based synchrony in primary auditory cortex and suggest that robust population coding of the sound envelope information present in animal vocalizations and speech can be reliably achieved even with indiscriminate pooling of cortical responses. These findings highlight the importance of firing rate dynamics in population-based sensory coding.NEW & NOTEWORTHY Fundamental questions remain about population coding in primary auditory cortex (A1). In particular, issues of spike timing in models of neural populations have been largely ignored. We find that spike-timing in response to sound envelope fluctuations is highly similar across neuron populations in A1. This property of shared envelope phase preference allows for a simple population model involving unweighted convergence of neuronal responses to classify amplitude modulation frequencies with high accuracy.

Extracellular voltage thresholds for maximizing information extraction in primate auditory cortex: implications for a brain computer interface.

Objective. Research by Oby (2016J. Neural. Eng.13036009) demonstrated that the optimal threshold for extracting information from visual and motor cortices may differ from the optimal threshold for identifying single neurons via spike sorting methods. The optimal threshold for extracting information from auditory cortex has yet to be identified, nor has the optimal temporal scale for representing auditory cortical activity. Here, we describe a procedure to jointly optimize the extracellular threshold and bin size with respect to the decoding accuracy achieved by a linear classifier for a diverse set of auditory stimuli.Approach. We used linear multichannel arrays to record extracellular neural activity from the auditory cortex of awake squirrel monkeys passively listening to both simple and complex sounds. We executed a grid search of the coordinate space defined by the voltage threshold (in units of standard deviation) and the bin size (in units of milliseconds), and computed decoding accuracy at each point.Main results. The optimal threshold for information extraction was consistently near two standard deviations below the voltage trace mean, which falls significantly below the range of three to five standard deviations typically used as inputs to spike sorting algorithms in basic research and in brain-computer interface (BCI) applications. The optimal binwidth was minimized at the optimal voltage threshold, particularly for acoustic stimuli dominated by temporally dynamic features, indicating that permissive thresholding permits readout of cortical responses with temporal precision on the order of a few milliseconds.Significance. The improvements in decoding accuracy we observed for optimal readout parameters suggest that standard thresholding methods substantially underestimate the information present in auditory cortical spiking patterns. The fact that optimal thresholds were relatively low indicates that local populations of cortical neurons exhibit high temporal coherence that could be leveraged in service of future auditory BCI applications.

Amplitude modulation coding in awake mice and squirrel monkeys.

Both mice and primates are used to model the human auditory system. The primate order possesses unique cortical specializations that govern auditory processing. Given the power of molecular and genetic tools available in the mouse model, it is essential to understand the similarities and differences in auditory cortical processing between mice and primates. To address this issue, we directly compared temporal encoding properties of neurons in the auditory cortex of awake mice and awake squirrel monkeys (SQMs). Stimuli were drawn from a sinusoidal amplitude modulation (SAM) paradigm, which has been used previously both to characterize temporal precision and to model the envelopes of natural sounds. Neural responses were analyzed with linear template-based decoders. In both species, spike timing information supported better modulation frequency discrimination than rate information, and multiunit responses generally supported more accurate discrimination than single-unit responses from the same site. However, cortical responses in SQMs supported better discrimination overall, reflecting superior temporal precision and greater rate modulation relative to the spontaneous baseline and suggesting that spiking activity in mouse cortex was less strictly regimented by incoming acoustic information. The quantitative differences we observed between SQM and mouse cortex support the idea that SQMs offer advantages for modeling precise responses to fast envelope dynamics relevant to human auditory processing. Nevertheless, our results indicate that cortical temporal processing is qualitatively similar in mice and SQMs and thus recommend the mouse model for mechanistic questions, such as development and circuit function, where its substantial methodological advantages can be exploited. NEW & NOTEWORTHY To understand the advantages of different model organisms, it is necessary to directly compare sensory responses across species. Contrasting temporal processing in auditory cortex of awake squirrel monkeys and mice, with parametrically matched amplitude-modulated tone stimuli, reveals a similar role of timing information in stimulus encoding. However, disparities in response precision and strength suggest that anatomical and biophysical differences between squirrel monkeys and mice produce quantitative but not qualitative differences in processing strategy.

Cluster-based analysis improves predictive validity of spike-triggered receptive field estimates.

Spectrotemporal receptive field (STRF) characterization is a central goal of auditory physiology. STRFs are often approximated by the spike-triggered average (STA), which reflects the average stimulus preceding a spike. In many cases, the raw STA is subjected to a threshold defined by gain values expected by chance. However, such correction methods have not been universally adopted, and the consequences of specific gain-thresholding approaches have not been investigated systematically. Here, we evaluate two classes of statistical correction techniques, using the resulting STRF estimates to predict responses to a novel validation stimulus. The first, more traditional technique eliminated STRF pixels (time-frequency bins) with gain values expected by chance. This correction method yielded significant increases in prediction accuracy, including when the threshold setting was optimized for each unit. The second technique was a two-step thresholding procedure wherein clusters of contiguous pixels surviving an initial gain threshold were then subjected to a cluster mass threshold based on summed pixel values. This approach significantly improved upon even the best gain-thresholding techniques. Additional analyses suggested that allowing threshold settings to vary independently for excitatory and inhibitory subfields of the STRF resulted in only marginal additional gains, at best. In summary, augmenting reverse correlation techniques with principled statistical correction choices increased prediction accuracy by over 80% for multi-unit STRFs and by over 40% for single-unit STRFs, furthering the interpretational relevance of the recovered spectrotemporal filters for auditory systems analysis.

Effects of Signal-to-Noise Ratio on Auditory Cortical Frequency Processing.

The neural mechanisms that support the robust processing of acoustic signals in the presence of background noise in the auditory system remain largely unresolved. Psychophysical experiments have shown that signal detection is influenced by the signal-to-noise ratio (SNR) and the overall stimulus level, but this relationship has not been fully characterized. We evaluated the neural representation of frequency in rat primary auditory cortex by constructing tonal frequency response areas (FRAs) in primary auditory cortex for different SNRs, tone levels, and noise levels. We show that response strength and selectivity for frequency and sound level depend on interactions between SNRs and tone levels. At low SNRs, jointly increasing the tone and noise levels reduced firing rates and narrowed FRA bandwidths; at higher SNRs, however, increasing the tone and noise levels increased firing rates and expanded bandwidths, as is usually seen for FRAs obtained without background noise. These changes in frequency and intensity tuning decreased tone level and tone frequency discriminability at low SNRs. By contrast, neither response onset latencies nor noise-driven steady-state firing rates meaningfully interacted with SNRs or overall sound levels. Speech detection performance in humans was also shown to depend on the interaction between overall sound level and SNR. Together, these results indicate that signal processing difficulties imposed by high noise levels are quite general and suggest that the neurophysiological changes we see for simple sounds generalize to more complex stimuli. SIGNIFICANCE STATEMENT: Effective processing of sounds in background noise is an important feature of the mammalian auditory system and a necessary feature for successful hearing in many listening conditions. Even mild hearing loss strongly affects this ability in humans, seriously degrading the ability to communicate. The mechanisms involved in achieving high performance in background noise are not well understood. We investigated the effects of SNR and overall stimulus level on the frequency tuning of neurons in rat primary auditory cortex. We found that the effects of noise on frequency selectivity are not determined solely by the SNR but depend also on the levels of the foreground tones and background noise. These observations can lead to improvement in therapeutic approaches for hearing-impaired patients.

Modulation-frequency-specific adaptation in awake auditory cortex.

Amplitude modulations are fundamental features of natural signals, including human speech and nonhuman primate vocalizations. Because natural signals frequently occur in the context of other competing signals, we used a forward-masking paradigm to investigate how the modulation context of a prior signal affects cortical responses to subsequent modulated sounds. Psychophysical "modulation masking," in which the presentation of a modulated "masker" signal elevates the threshold for detecting the modulation of a subsequent stimulus, has been interpreted as evidence of a central modulation filterbank and modeled accordingly. Whether cortical modulation tuning is compatible with such models remains unknown. By recording responses to pairs of sinusoidally amplitude modulated (SAM) tones in the auditory cortex of awake squirrel monkeys, we show that the prior presentation of the SAM masker elicited persistent and tuned suppression of the firing rate to subsequent SAM signals. Population averages of these effects are compatible with adaptation in broadly tuned modulation channels. In contrast, modulation context had little effect on the synchrony of the cortical representation of the second SAM stimuli and the tuning of such effects did not match that observed for firing rate. Our results suggest that, although the temporal representation of modulated signals is more robust to changes in stimulus context than representations based on average firing rate, this representation is not fully exploited and psychophysical modulation masking more closely mirrors physiological rate suppression and that rate tuning for a given stimulus feature in a given neuron's signal pathway appears sufficient to engender context-sensitive cortical adaptation.

Representation of loudness in the auditory cortex.

Changes in stimulus intensity are reflected in changes in the fundamental perceptual attribute of loudness. Stimulus intensity changes also profoundly impact the evoked neural responses throughout the auditory system. A fundamental question is how measurements of neural activity, from the single-neuron level to mass-activity metrics such as functional magnetic resonance imaging or magnetoencephalography, reflect the physical properties of stimulus intensity as opposed to perceived loudness. In this chapter we discuss findings from psychophysics and animal neurophysiology as well as human brain activity measurements to clarify our current understanding of the neural mechanisms that contribute to the perceptual correlate of stimulus intensity.

Diverse cortical codes for scene segmentation in primate auditory cortex.

The temporal coherence of amplitude fluctuations is a critical cue for segmentation of complex auditory scenes. The auditory system must accurately demarcate the onsets and offsets of acoustic signals. We explored how and how well the timing of onsets and offsets of gated tones are encoded by auditory cortical neurons in awake rhesus macaques. Temporal features of this representation were isolated by presenting otherwise identical pure tones of differing durations. Cortical response patterns were diverse, including selective encoding of onset and offset transients, tonic firing, and sustained suppression. Spike train classification methods revealed that many neurons robustly encoded tone duration despite substantial diversity in the encoding process. Excellent discrimination performance was achieved by neurons whose responses were primarily phasic at tone offset and by those that responded robustly while the tone persisted. Although diverse cortical response patterns converged on effective duration discrimination, this diversity significantly constrained the utility of decoding models referenced to a spiking pattern averaged across all responses or averaged within the same response category. Using maximum likelihood-based decoding models, we demonstrated that the spike train recorded in a single trial could support direct estimation of stimulus onset and offset. Comparisons between different decoding models established the substantial contribution of bursts of activity at sound onset and offset to demarcating the temporal boundaries of gated tones. Our results indicate that relatively few neurons suffice to provide temporally precise estimates of such auditory "edges," particularly for models that assume and exploit the heterogeneity of neural responses in awake cortex.

Encoding frequency contrast in primate auditory cortex.

Changes in amplitude and frequency jointly determine much of the communicative significance of complex acoustic signals, including human speech. We have previously described responses of neurons in the core auditory cortex of awake rhesus macaques to sinusoidal amplitude modulation (SAM) signals. Here we report a complementary study of sinusoidal frequency modulation (SFM) in the same neurons. Responses to SFM were analogous to SAM responses in that changes in multiple parameters defining SFM stimuli (e.g., modulation frequency, modulation depth, carrier frequency) were robustly encoded in the temporal dynamics of the spike trains. For example, changes in the carrier frequency produced highly reproducible changes in shapes of the modulation period histogram, consistent with the notion that the instantaneous probability of discharge mirrors the moment-by-moment spectrum at low modulation rates. The upper limit for phase locking was similar across SAM and SFM within neurons, suggesting shared biophysical constraints on temporal processing. Using spike train classification methods, we found that neural thresholds for modulation depth discrimination are typically far lower than would be predicted from frequency tuning to static tones. This "dynamic hyperacuity" suggests a substantial central enhancement of the neural representation of frequency changes relative to the auditory periphery. Spike timing information was superior to average rate information when discriminating among SFM signals, and even when discriminating among static tones varying in frequency. This finding held even when differences in total spike count across stimuli were normalized, indicating both the primacy and generality of temporal response dynamics in cortical auditory processing.

Spectral context affects temporal processing in awake auditory cortex.

Amplitude modulation encoding is critical for human speech perception and complex sound processing in general. The modulation transfer function (MTF) is a staple of auditory psychophysics, and has been shown to predict speech intelligibility performance in a range of adverse listening conditions and hearing impairments, including cochlear implant-supported hearing. Although both tonal and broadband carriers have been used in psychophysical studies of modulation detection and discrimination, relatively little is known about differences in the cortical representation of such signals. We obtained MTFs in response to sinusoidal amplitude modulation (SAM) for both narrowband tonal carriers and two-octave bandwidth noise carriers in the auditory core of awake squirrel monkeys. MTFs spanning modulation frequencies from 4 to 512 Hz were obtained using 16 channel linear recording arrays sampling across all cortical laminae. Carrier frequency for tonal SAM and center frequency for noise SAM was set at the estimated BF for each penetration. Changes in carrier type affected both rate and temporal MTFs in many neurons. Using spike discrimination techniques, we found that discrimination of modulation frequency was significantly better for tonal SAM than for noise SAM, though the differences were modest at the population level. Moreover, spike trains elicited by tonal and noise SAM could be readily discriminated in most cases. Collectively, our results reveal remarkable sensitivity to the spectral content of modulated signals, and indicate substantial interdependence between temporal and spectral processing in neurons of the core auditory cortex.

Transformation of temporal processing across auditory cortex of awake macaques.

The anatomy and connectivity of the primate auditory cortex has been modeled as a core region receiving direct thalamic input surrounded by a belt of secondary fields. The core contains multiple tonotopic fields (including the primary auditory cortex, AI, and the rostral field, R), but available data only partially address the degree to which those fields are functionally distinct. This report, based on single-unit recordings across four hemispheres in awake macaques, argues that the functional organization of auditory cortex is best understood in terms of temporal processing. Frequency tuning, response threshold, and strength of activation are similar between AI and R, validating their inclusion as a unified core, but the temporal properties of the fields clearly differ. Onset latencies to pure tones are longer in R (median, 33 ms) than in AI (20 ms); moreover, synchronization of spike discharges to dynamic modulations of stimulus amplitude and frequency, similar to those present in macaque and human vocalizations, suggest distinctly different windows of temporal integration in AI (20-30 ms) and R (100 ms). Incorporating data from the adjacent auditory belt reveals that the divergence of temporal properties within the core is in some cases greater than the temporal differences between core and belt.

Temporal codes for amplitude contrast in auditory cortex.

The encoding of sound level is fundamental to auditory signal processing, and the temporal information present in amplitude modulation is crucial to the complex signals used for communication sounds, including human speech. The modulation transfer function, which measures the minimum detectable modulation depth across modulation frequency, has been shown to predict speech intelligibility performance in a range of adverse listening conditions and hearing impairments, and even for users of cochlear implants. We presented sinusoidal amplitude modulation (SAM) tones of varying modulation depths to awake macaque monkeys while measuring the responses of neurons in the auditory core. Using spike train classification methods, we found that thresholds for modulation depth detection and discrimination in the most sensitive units are comparable to psychophysical thresholds when precise temporal discharge patterns rather than average firing rates are considered. Moreover, spike timing information was also superior to average rate information when discriminating static pure tones varying in level but with similar envelopes. The limited utility of average firing rate information in many units also limited the utility of standard measures of sound level tuning, such as the rate level function (RLF), in predicting cortical responses to dynamic signals like SAM. Response modulation typically exceeded that predicted by the slope of the RLF by large factors. The decoupling of the cortical encoding of SAM and static tones indicates that enhancing the representation of acoustic contrast is a cardinal feature of the ascending auditory pathway.

Representation of dynamic interaural phase difference in auditory cortex of awake rhesus macaques.

Neurons in auditory cortex of awake primates are selective for the spatial location of a sound source, yet the neural representation of the binaural cues that underlie this tuning remains undefined. We examined this representation in 283 single neurons across the low-frequency auditory core in alert macaques, trained to discriminate binaural cues for sound azimuth. In response to binaural beat stimuli, which mimic acoustic motion by modulating the relative phase of a tone at the two ears, these neurons robustly modulate their discharge rate in response to this directional cue. In accordance with prior studies, the preferred interaural phase difference (IPD) of these neurons typically corresponds to azimuthal locations contralateral to the recorded hemisphere. Whereas binaural beats evoke only transient discharges in anesthetized cortex, neurons in awake cortex respond throughout the IPD cycle. In this regard, responses are consistent with observations at earlier stations of the auditory pathway. Discharge rate is a band-pass function of the frequency of IPD modulation in most neurons (73%), but both discharge rate and temporal synchrony are independent of the direction of phase modulation. When subjected to a receiver operator characteristic analysis, the responses of individual neurons are insufficient to account for the perceptual acuity of these macaques in an IPD discrimination task, suggesting the need for neural pooling at the cortical level.

Dynamics of tuning in the Fourier domain.

Neurons in primary visual cortex (area V1) are jointly tuned to the orientation and spatial frequency of sinusoidal stimuli (the Fourier domain). The role that suppressive mechanisms play in shaping the tuning and dynamics of cortical responses remains the subject of debate. Here we used subspace reverse correlation to study the relationship between suppression by nonoptimal stimuli, the spectral-temporal separability of the responses, and their persistence in time. Two clear relationships emerged from our data. First, cells with inseparable responses were often accompanied by suppression to nonpreferred stimuli, while separable responses showed mostly enhancement by their preferred stimuli. Second, inseparable responses were characterized by a longer persistence in time compared with those with separable dynamics. A parametric model that assumes the additive combination of separable enhancement and suppression signals, with suppression constrained to be low-pass in spatial frequency and untuned for orientation, explained the data well. These new findings, in addition to an established correlation between selectivity and suppression for nonoptimal stimuli, clarify how the dynamics and selectivity of cortical responses are shaped by suppressive signals and how their interplay generates the large diversity of responses observed in primary visual cortex.

Dynamic amplitude coding in the auditory cortex of awake rhesus macaques.

In many animals, the information most important for processing communication sounds, including speech, consists of temporal envelope cues below approximately 20 Hz. Physiological studies, however, have typically emphasized the upper limits of modulation encoding. Responses to sinusoidal AM (SAM) are generally summarized by modulation transfer functions (MTFs), which emphasize tuning to modulation frequency rather than the representation of the instantaneous stimulus amplitude. Unfortunately, MTFs fail to capture important but nonlinear aspects of amplitude coding in the central auditory system. We focus on an alternative data representation, the modulation period histogram (MPH), which depicts the spike train folded on the modulation period of the SAM stimulus. At low modulation frequencies, the fluctuations of stimulus amplitude in decibels are robustly encoded by the cycle-by-cycle response dynamics evident in the MPH. We show that all of the parameters that define a SAM stimulus--carrier frequency, carrier level, modulation frequency, and modulation depth--are reflected in the shape of cortical MPHs. In many neurons that are nonmonotonically tuned for sound amplitude, the representation of modulation frequency is typically sacrificed to preserve the mapping between the instantaneous discharge rate and the instantaneous stimulus amplitude, resulting in two response modes per modulation cycle. This behavior, as well as the relatively poor tuning of cortical MTFs, suggests that auditory cortical neurons are not well suited for operating as a "modulation filterbank." Instead, our results suggest that <20 Hz, the processing of modulated signals is better described as envelope shape discrimination rather than modulation frequency extraction.

The operating point of the cortex: neurons as large deviation detectors.

Spiking neurons translate analog intracellular variables into a sequence of action potentials. A simplified model of this transformation is one in which an underlying "generator potential," representing a measure of overall neuronal drive, is passed through a static nonlinearity to produce an instantaneous firing rate. An important question is how adaptive mechanisms adjust the mean and SD of the generator potential to define an "operating point" that controls spike generation. In early sensory pathways adaptation has been shown to rescale the generator potential to maximize the amount of transmitted information. In contrast, we demonstrate that the operating point in the cortex is tuned so that cells respond only when the generator potential executes a large excursion above its mean value. The distance from the mean of the generator potential to spike threshold is, on average, 1 SD of the ongoing activity. Signals above threshold are amplified linearly and do not reach saturation. The operating point is adjusted dynamically so that it remains relatively invariant despite changes in stimulus contrast. We conclude that the operating regimen of the cortex is suitable for the detection of signals in background noise and for enhancing the selectivity of spike responses relative to those of the generator potential (the so-called "iceberg effect"), but not to maximize the transmission of total information.

Effect of behavioral context on representation of a spatial cue in core auditory cortex of awake macaques.

Primary auditory cortex plays a crucial role in spatially directed behavior, but little is known about the effect of behavioral state on the neural representation of spatial cues. Macaques were trained to discriminate binaural cues to sound localization, eventually allowing measurement of thresholds comparable to human hearing. During behavior and passive listening, single units in low-frequency auditory cortex showed robust and consistent tuning to interaural phase difference (IPD). In most neurons, behavior exerted an effect on peak discharge rate (58% increased, 13% decreased), but this was not accompanied by a detectable shift in the best IPD of any cell. Neurometric analysis revealed a difference in discriminability between the behaving and passive condition in half of the sample (52%), but steepening of the neurometric function (29%) was only slightly more common than flattening (23%). This suggests that performance of a discrimination task does not necessarily confer an advantage in understanding the representation of the spatial cue in primary auditory cortex but nevertheless revealed some physiological effects. These results suggest that responses observed during passive listening provide a valid representation of neuronal response properties in core auditory cortex.

Dynamics of receptive field size in primary visual cortex.

Recent studies have shown that the initial responses evoked by a stimulus in neurons of primary visual cortex are dominated by low spatial frequency information in the image, whereas finer spatial scales dominate later in the response. Such phenomena could arise from the dynamics of receptive field (RF) size at early stages of cortical processing. We measured changes in RF size in simple cells recorded from the primary visual cortex of anesthetized macaques by measuring their first-order spatio-temporal kernels and fitting them with two-dimensional Gabor functions at different time slices. We found that the width and length of the RF envelope and the period of the carrier tend to decrease during the time-course of the response. The most pronounced changes are seen in the width and spatial period of the RFs, which decrease by 15% during the central 20 ms of the response. These results show a novel form of spatio-temporal inseparability in simple cells and are consistent with the notion of a coarse-to-fine processing of information in early visual cortex.

Organochlorines and mercury in waterfowl harvested in Canada.

Samples of breast muscle from 32 species of waterfowl collected from 123 sites across Canada were analyzed for chlorobenzenes (CBz), chlordane-related compounds (CHL), hexachlorocyclohexanes (HCH), DDT, mirex, dieldrin, PCBs and mercury. SigmaDDT, SigmaCBz and SigmaPCB were the compounds most frequently found above trace levels. SigmaHCH and SigmaMirex were detected the least often. Mercury was detected in all of the mergansers, over 50% of dabbling, bay and sea ducks, and in less than 2% of the geese analysed. The highest levels of contaminants were generally found in birds feeding at higher trophic levels such as sea ducks and mergansers. With the exception of a few samples of mergansers and long-tailed ducks from eastern Canada, which contained SigmaPCB concentrations of 1.0-2.4 mg kg(-1), SigmaPCB levels were less than 1 mg kg(-1) wet weight. Only one merganser from eastern Canada had a SigmaDDT concentration (2.6 mg kg(-1) ww) which was greater than 1 mg kg(-1) ww. The highest SigmaCHL (0.10 mg kg(-1) ww) was also found in mergansers from eastern Canada. Levels of total mercury in breast muscle were either low (< 1 mg kg(-1) ww) or below detection limits with the exception of a few samples of mergansers from eastern Canada which contained mercury concentrations of 1.0-1.5 mg kg(-1) ww. Health Canada determined that the organochlorine and mercury levels found in samples of breast muscle of ducks and geese analysed in this study did not pose a health hazard to human consumers and therefore these waterfowl were safe to eat.

Organochlorines and trace elements in upland game birds harvested in Canada.

American woodcock, common snipe, two species of ptarmigan (willow and rock), and three species of grouse (ruffed, spruce, and sharp-tailed) were collected from 21 locations between 1991 and 1994 as part of a nationwide survey of contaminants in game birds harvested in Canada. Breast muscle was analysed for organochlorines (chlorobenzenes, hexachlorocyclohexanes, chlordane-related compounds, DDT metabolites, mirex, dieldrin, and PCBs) and trace elements (Hg, Cd, Se, and As). The concentrations of organochlorine compounds measured in breast muscle of the birds in this survey were very low. Median concentrations for the organochlorines measured were less than 0.001 mg kg-1 in breast muscle of all ptarmigan and grouse species sampled. Only SigmaPCB, SigmaDDT and SigmaCHL were found above trace levels in breast muscle of all of the woodcock, and only SigmaPCB and SigmaDDT were found above trace levels in the snipe. Total Hg was detected only in the woodcock and the snipe whereas Se was above detection levels in all samples of all species except one sample pool of willow ptarmigan from Nunavut. Cadmium was detected in all species except for the spruce and sharp-tailed grouse with the highest overall concentrations found in the ptarmigan species. Arsenic was detected in all of the woodcock and snipe samples but was not found in any of the grouse samples. The highest Hg and As concentrations were both found in snipe sampled from southeastern New Brunswick. Concentrations of As, Hg and Se measured in breast muscle of the birds in this survey were below toxicological threshold levels found in the literature. Concentrations of Cd, however, were elevated in some birds, particularly in willow ptarmigan from the Yukon. Health Canada determined that the organochlorine and trace element levels found in samples of breast muscle of game birds analysed in this study did not pose a health hazard to human consumers and therefore these birds were safe to eat.