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The general and comparative biology of gonadotropin-inhibitory hormone (GnIH).

The decapeptide gonadotropin-releasing hormone (GnRH) is the primary factor responsible for the hypothalamic control of gonadotropin secretion. Gonadal sex steroids and inhibin inhibit gonadotropin secretion via feedback from the gonads, but a neuropeptide inhibitor of gonadotropin secretion was, until recently, unknown in vertebrates. In 2000, we identified a novel hypothalamic dodecapeptide that inhibits gonadotropin release in cultured quail pituitaries and termed it gonadotropin-inhibitory hormone (GnIH). To elucidate the mode of action of GnIH, we then identified a novel G protein-coupled receptor for GnIH in quail. The GnIH receptor possesses seven transmembrane domains and specifically binds to GnIH. The GnIH receptor is expressed in the pituitary and several brain regions including the hypothalamus. These results indicate that GnIH acts directly on the pituitary via GnIH receptor to inhibit gonadotropin release. GnIH may also act on the hypothalamus to inhibit GnRH release. To demonstrate the functional significance of GnIH and its potential role as a key regulatory neuropeptide in avian reproduction, we investigated GnIH actions on gonadal development and maintenance in quail. Chronic treatment with GnIH inhibited gonadal development and maintenance by decreasing gonadotropin synthesis and release. GnIH was also found in the hypothalamus of other avian species including sparrows and chickens and also inhibited gonadotropin synthesis and release. The pineal hormone melatonin may be a key factor controlling GnIH neural function, since quail GnIH neurons express melatonin receptor and melatonin treatment stimulates the expression of GnIH mRNA and mature GnIH peptide. Thus, GnIH is capable of transducing photoperiodic information via changes in the melatonin signal, thereby influencing the reproductive axis. It is concluded that GnIH, a newly discovered hypothalamic neuropeptide, is a key factor controlling avian reproduction. The discovery of avian GnIH opens a new research field in reproductive neuroendocrinology.

Amino Acid Sequence↗

Testosterone reduces responsiveness to nociceptive stimuli in a wild bird.

The hormone testosterone (T) is involved in the control of aggressive behavior in male vertebrates. T enhances the frequency and intensity of aggressive behaviors during competitive interactions among males. By promoting high-intensity aggression, T also increases the risk of injury and presumably the perception of painful stimuli. However, perception of painful stimuli during fights could counteract the expression of further aggressive behavior. We therefore hypothesize that one function of T during aggressive interactions is to reduce nociception (pain sensitivity). Here, we experimentally document that T indeed reduces behavioral responsiveness to a thermal painful stimulus in captive male house sparrows (Passer domesticus). Skin nociception was quantified by foot immersion into a hot water bath, a benign thermal stimulus. Males treated with exogenous testosterone left their foot longer in hot water than control birds. Conversely, males in which the physiological actions of testosterone were pharmacologically blocked withdrew their foot faster than control birds. Testosterone might exert its effects on pain sensitivity through conversion into estradiol in the dorsal horn of the spinal cord. Decreased nociception during aggressive encounters may promote the immediate and future willingness of males to engage in high-intensity fights.

Aggression↗

The use of fluorescein isothiocyanate labelled lectins for immunohistochemical demonstration of saccharides.

Cryostat sections of various tissues from man, rat, mice, hamster, rabbit, guinea pig, sparrow and carp were examined by fluorescence microscopy after incubation with fluorescein isothiocyanate labelled Concanavalin A and Lens culinaris lectin. In the same tissue of various species no basic differences in the pattern of fluorescence were observed. In general, connective tissue structures were stained by both lectins in the same degree, whereas by fluorescein isothiocyanate labelled Concanavalin A additionally a varying staining of the cell surface and of cytoplasmic structures occurred. The results show the general presence of Concanavalin A and Lens culinaris lectin receptors in normal animal tissues.

Animals↗

Short-term fasting affects locomotor activity, corticosterone, and corticosterone binding globulin in a migratory songbird.

Unpredictable events such as severe storms lead to an increase in circulating corticosterone (CORT) in breeding birds. This increase is often accompanied by elevations in foraging and irruptive behavior. We were interested in determining if acute food restriction (such as might occur during inclement weather) is a sufficient cue to elicit an increase in locomotor activity, increase CORT secretion, and/or decrease circulating levels of corticosterone binding globulin (CBG) in white-crowned sparrows (Zonotrichia leucophrys gambelii). Male Z.l. gambelii were housed individually in environmental chambers on long days (LD 20:4) to simulate breeding season daylength. Birds were fed ad libitum, and on select days, food was removed 2 h after lights on (fasted treatment), or was removed and replaced (control). We analyzed CORT and CBG levels after 1, 2, 6, 22 (lights on), and 23 h under fasted and control conditions. We also measured activity during the 23-h experiment. Activity levels were increased under fasted conditions during the daytime relative to control conditions, but activity levels did not differ between treatments during the night. Fasting as little as 1, 2, and 6 h significantly increased total CORT levels above baseline (control), although after 22 h, total CORT levels under fasted conditions matched those under control conditions. Plasma CBG decreased after the 22-h fast, and remained low after the 23-h fast. This change was sufficient to significantly elevate free CORT levels in fasted birds relative to ad libitum food conditions, despite the lack of difference in total CORT levels.

Animal Migration↗

Three-dimensional analysis of avian song control nuclei.

Analysis of seasonal and developmental changes in the morphology of avian song control nuclei has traditionally been performed using two-dimensional (2-D) cross-sectional traces from brain sections. This method, although reliable, does not encompass the possibility that subdivisions of a nucleus might change in size to different degrees. Three-dimensional (3-D) analysis of song nuclei under different conditions could provide insight on this issue. This approach could also be of value in guiding and evaluating the use of lesions to study the functions of subdivisions of song nuclei. We used customized computer software to produce 3-D images of song nuclei from 2-D brain sections of spotted towhees (Pipilo maculatus) in different hormonal status, and from Gambel's white-crowned sparrows (Zonotrichia leucophrys gambelii) with unilateral lesions of the higher vocal center (HVc). 3-D images show that some sub-regions of song nuclei indeed change in size to a greater extent than others. 3-D analysis of HVc lesions provides a clearer view of the size and shape of the lesion site within the target nucleus and relative to the surrounding tissue. Used in conjunction with 2-D analysis, the 3-D method will aid investigations of the song system and contribute to the understanding of its regulation by hormones.

Animals↗

Isolation of avian pneumovirus from mallard ducks that is genetically similar to viruses isolated from neighboring commercial turkeys.

Our earlier studies demonstrating avian pneumovirus (APV) RNA in wild geese, sparrows, swallows, starlings and mallard ducks suggested that wild birds might be involved in the circulation of APV in the United States. To determine whether turkey virus can be transmitted to the free flying birds, we placed APV-negative mallard ducks next to a turkey farm experiencing a severe APV outbreak and in an area with a large population of waterfowls. The sentinel ducks did not develop clinical APV disease but infectious APV (APV/MN-12) was recovered from choanal swabs after 2 weeks, and anti-APV antibodies detected after 4 weeks. Four APV isolates recovered from the neighboring turkeys that were experiencing an APV outbreak at the same time shared 95-99% nucleotide identity and 97-99% predicted amino acid identity with the duck isolate. In addition experimental infection of turkey poults with APV/MN-12 resulted in detection of viral RNA in nasal turbinates and APV-specific IgG in serum. These results indicate that the APV isolates from turkeys and ducks shared a common source, and the viruses from different avian species can cross-infect.

Amino Acid Sequence↗

Heavy metals and metalloids in egg contents and eggshells of passerine birds from Arizona.

Concentrations of inorganic elements were determined in eggs of passerine birds including the endangered southwestern willow flycatcher (Empidonax traillii extimus) from four regions in Arizona. The main aim of the study was to determine the distribution of metals in egg contents and eggshells, with emphasis on the deposition of Sr in eggshells. Seventy eggs of 11 passerine species were collected at four nesting locations during 2000. Aluminum, Ba, Cr, Cu, Mn, Se, Sr, and Zn, were detected primarily in egg contents of all bird species. Arsenic, Ni, Pb, and V were detected primarily in eggshells. A proportion of most inorganic elements accumulated in the eggshell. Concentrations of Ba, Cu, Mn, Se, Sr, and Zn in egg contents and As, Ba, Cu, and V in eggshells of yellow-breasted chats (Icteria virens) were similar among locations. However, concentrations of Mn, Ni, Sr, and Zn in eggshells were significant different among locations. Except for Cu, Mn, Se, and Zn, concentrations of inorganic elements were 2-35 times greater in eggshells than in eggs. Most concentrations of metals and metalloids in eggs and eggshells of all the bird species were below levels known to affect reproduction or that have other deleterious effects. However, I found somewhat elevated concentrations of Sr in eggshells (highest MEAN=1505 microg/g dw, n=3) of yellow-breasted chats and willow flycatchers, and in egg contents of yellow warblers (Dendroica petechia) and song sparrows (Melospiza melodia). Whether current observed concentrations of Sr in eggshells are affecting nesting birds in Arizona remains to be determined. Strontium and other metals could be associated with lower hatching success in some areas. This study shows that a proportion of many inorganic elements accumulates in the eggshell and that the potential effects on the proper structure and functioning of the eggshell should not be ignored.

Animals↗

Daily and circadian fluctuation in 2-deoxy[(14)C]-glucose uptake in circadian and visual system structures of the chick brain: effects of exogenous melatonin.

Previous studies show that several structures of the house sparrow visual system are metabolically rhythmic, as determined by 2-deoxy[(14)C]glucose (2DG) uptake, and that these metabolic rhythms depend upon rhythmic melatonin in this species. In many species of birds, high affinity binding of 2[(125)I] iodomelatonin is widespread in the brain, especially in visual system structures. The present study asks whether 2DG uptake is similarly rhythmic in the chick brain and whether exogenous melatonin administration affects 2DG uptake. Chicks were injected with 2DG and sacrificed 1 h later. Their brains were removed and processed for 2DG autoradiography. Chicks were injected during the late day with melatonin or saline prior to the 2DG injection and brain processing. We found that the visual suprachiasmatic nucleus showed both daily and circadian differences in 2DG uptake. Six of seven visual structures displayed daily uptake changes, while only two structures showed circadian fluctuations. Melatonin affected daytime 2DG uptake within visual suprachiasmatic nucleus and ectostriatum only. These results indicate that the chick circadian system is involved in the regulation of energy metabolism in the visual system but that the role for pineal melatonin in that regulatory process is a subtle one.

Animals↗

Levels of categorization in visual recognition studied using functional magnetic resonance imaging.

BACKGROUND: Recent functional neuroimaging results implicate part of the ventral temporal lobe of the brain in face recognition, and have, together with neurophysiological findings, been used as evidence for a face-specific neural module in the brain. Experimental designs, however, have often failed to distinguish between the class of the object used as the stimulus (face or non-face) and the level of categorization at which the stimulus is recognized (the 'basic' level, such as 'bird', at which familiar objects are first recognized, or more subordinate levels - 'sparrow', for example - which require additional perceptual processing). We have used echo-planar functional magnetic resonance imaging to compare brain activation for the matching of non-face objects with subordinate-level and basic-level descriptors. RESULTS: The additional visual processing required to verify the subordinate level of a picture over its basic level was associated with activation of the fusiform and inferior temporal gyri (FIT) as well as the temporal poles. These areas correspond closely to those previously implicated in the processing of facial images. CONCLUSIONS: Our results indicate that areas of the ventral visual pathway that have been associated with face recognition are sensitive to manipulations of the categorization level of non-face objects. This idea offers an alternative to the dominant view that FIT may be organized according to conceptual categories, and our results establish the importance of manipulating task requirements when evaluating a 'neural module' hypothesis.

Brain Mapping↗

Relief of chronic neck and shoulder pain by manual acupuncture to tender points--a sham-controlled randomized trial.

OBJECTIVES: To compare the effects of real acupuncture to tender points for neck and shoulder pain and stiffness (Japanese: katakori) with those of sham acupuncture. DESIGN: Randomized-controlled trial. METHODS: Thirty-four volunteers from an acupuncture school with complaints of chronic pain and stiffness, who had no arm symptoms and gave informed consent, were randomly allocated to acupuncture or sham groups. Acupuncture or sham acupuncture was applied to the tender points once a week for 3 weeks. In the acupuncture group the acupuncture needle was inserted to the muscle, then the sparrow pecking technique was applied five times. Sham acupuncture was done without insertion of the needle. Dull pain and stiffness were evaluated by visual analog scale (VAS) before, and every 2 days after the first needling for 1 month. Pressure pain threshold on the tender points was measured before and after each treatment. RESULTS: There was no statistical difference of VAS scores between acupuncture and sham groups 9 days after the last treatment. However, the acupuncture group showed significant reduction of VAS scores immediately after and/or 1 day after the real acupuncture treatments (P<0.01). The effect tended to be prolonged after repeated treatment. Pressure pain thresholds tended to increase after real acupuncture treatment but not after sham acupuncture. CONCLUSIONS: Acupuncture applied to tender points appears to have short-term effects on neck and shoulder pain and stiffness, but this study was unable to demonstrate any long-term superiority over sham acupuncture.

Acupuncture Analgesia↗

Molecular phylogenetic analysis of Fringillidae, "New World nine-primaried oscines" (Aves: Passeriformes).

Systematic studies of Fringillidae have long been problematic due to their apparent recent and explosive diversification. We present phylogenetic hypotheses of 44 fringillids that represent the overall diversity of the family, based on 3.2 kb of mitochondrial DNA sequences, and phylogenetic analyses for a subset of fringillids based on new and published mitochondrial cytochrome b sequences. Monophyly of Fringillidae and its two constituent subfamilies, Fringillinae and Emberizinae, was consistently supported with the exceptions of Peucedramus being placed outside of Fringillinae and Euphonia being placed within Fringillinae instead of within Emberizinae. Within Emberizinae, Thraupini (tanagers), Cardinalini (cardinals and grosbeaks), and Emberizini (New World sparrows) did not form separate monophyletic groups. Our results indicate that Emberizinae consists of three clades, each with a different overall geographical distribution. Several taxa traditionally considered members of Thraupini fall outside of the thraupine clade, including the only North American genus, Piranga. Consequently, the thraupine clade includes only Neotropical species. Increasing evidence suggests that Fringillidae, often called "New World nine-primaried oscines," does not in fact have a New World origin.

Adenosine Triphosphatases↗

Longspurs and snow buntings: phylogeny and biogeography of a high-latitude clade (Calcarius).

Using complete cytochrome b sequence data, we determined that the genus Calcarius, as presently recognized, is paraphyletic. Calcarius plus Plectrophenax form a highly supported clade composed of two subclades, a "snow bunting" clade comprised of Plectrophenax plus Calcarius mccownii (formerly in the monotypic genus Rhynchophanes), and a "collared" longspur clade of Calcarius lapponicus, ornatus, and pictus. Contrary to conventional thought, Calcarius is not phylogenetically close to either Calamospiza or Emberiza. Unlike these two genera, the taxonomic affinities of Calcarius appear to lie outside of the sparrow (tribe Emberizini) assemblage. Calcarius appears to be a relatively old songbird lineage, originating between 4.2 and 6.2 million years ago. Within Calcarius, pictus and ornatus form a closely related sister pair (2.9% divergent), as do Calcarius nivalis and hyperboreus (0.18% divergent). The group (Calcarius, sensu lato) is inferred to have its origins at relatively high latitudes in the New World.

Alaska↗

Phylogeny of Passerida (Aves: Passeriformes) based on nuclear and mitochondrial sequence data.

Passerida is a monophyletic group of oscine passerines that includes almost 3500 species (about 36%) of all bird species in the world. The current understanding of higher-level relationships within Passerida is based on DNA-DNA hybridizations [C.G. Sibley, J.E. Ahlquist, Phylogeny and Classification of Birds, 1990, Yale University Press, New Haven, CT]. Our results are based on analyses of 3130 aligned nucleotide sequence data obtained from 48 ingroup and 13 outgroup genera. Three nuclear genes were sequenced: c-myc (498-510 bp), RAG-1 (930 bp), and myoglobin (693-722 bp), as well one mitochondrial gene; cytochrome b (879 bp). The data were analysed by parsimony, maximum-likelihood, and Bayesian inference. The African rockfowl and rockjumper are found to constitute the deepest branch within Passerida, but relationships among the other taxa are poorly resolved--only four major clades receive statistical support. One clade corresponds to Passeroidea of [C.G. Sibley, B.L. Monroe, Distribution and Taxonomy of Birds of the World, 1990, Yale University Press, New Haven, CT] and includes, e.g., flowerpeckers, sunbirds, accentors, weavers, estrilds, wagtails, finches, and sparrows. Starlings, mockingbirds, thrushes, Old World flycatchers, and dippers also group together in a clade corresponding to Muscicapoidea of Sibley and Monroe [op. cit.]. Monophyly of their Sylvioidea could not be corroborated--these taxa falls either into a clade with wrens, gnatcatchers, and nuthatches, or one with, e.g., warblers, bulbuls, babblers, and white-eyes. The tits, penduline tits, and waxwings belong to Passerida but have no close relatives among the taxa studied herein.

Animals↗

Editorial.

In this, the 11th Annual Research Review, I have been pleased to work with an outstanding group of contributors. As in past issues of the Annual Research Review the aim is to provide our readers with reviews that update both current knowledge and research findings. Authors are asked to be selective, rather than comprehensive, in their coverage as they identify the issues that they feel are particularly important for future research. I am grateful not only to the authors but to the numerous referees who provided critiques of each paper. In the first paper in this issue David Skuse provides an update on the relevance of behavioural neuroscience to child psychopathology. This paper provides a thoughtful review of the findings of the past decade and outlines possible directions for future research developments; it appears that we are poised for a major explosion of knowledge in this area. In the second paper Robin Chapman provides a very useful review of recent research on language development. This paper provides an update of Dorothy Bishop's earlier review of the topic and illustrates the considerable progress made since the time of that review. In the third paper Eilish Gilvarry summarises recent research on substance abuse in young people. This review covers recent changes in trends and patterns of substance abuse, aspects of risk and comorbidity, and treatment. Brown and colleagues then review recent work on children and adolescents with HIV and AIDS; this global health problem presents unique issues relative both to research and intervention. Danya Glaser then provides an overview of recent work on child abuse and neglect and the brain; the attempt to bring the various perspectives of neuroscience together on this topic is particularly timely and appropriate. Finally, Sparrow and Davis provide an overview of recent advances in the assessment of intelligence. This paper provides a helpful summary of current perspectives on the assessment of intelligence; the review of instruments will be of particular interest to our readers. For the 12th edition of the Annual Research Review we anticipate coverage of the following topics: intersubjectivity, reading disability, longitudinal approaches to developmental data, mental retardation, conduct disorder, and psychopharmacology.

Journal Article↗

Retinotopic representation of the bifoveate eye of the kestrel (Falco spraverius) on the optic tectum.

Like many diurnal raptors, the American kestrel or sparrow hawk, Falco sparverius, possesses two foveae in each eye. In this study, we used fundus photography and reversed ophthalmoscopy to plot the projection of these foveae onto the visual field, together with other retinal landmarks such as the pecten and ora terminali. From such data, it was determined that the central monocular fovea (CMF) and temporal binocular fovea (TBF) were separated by 36 deg and that the kestrel has 58 deg of binocular overlap when the eyes are appropriately converged. Single-cell and multi-unit recordings were used to systematically explore the tectal surface and map receptive fields (RF) onto a hemispheric screen. A retinotopic map of the tectal surface was produced from such data and revealed an expanded representation for each fovea on the tectum and a systematic increase in RF size from fovea to periphery. The functional significance of this organization is discussed.

Animals↗

Secretory phospholipase A2 and lipoprotein lipase enhance 15-lipoxygenase-induced enzymic and nonenzymic lipid peroxidation in low-density lipoproteins.

The oxidation of low-density lipoprotein (LDL) is thought to contribute to atherogenesis. 15-Lipoxygenase (15LO) induces LDL oxidation, and phospholipase A2 enhances this process [Sparrow, C. P. , Parthasarathy, S., and Steinberg, D. (1988) J. LipidRes. 29, 745-753]. As the underlying mechanism of the enhancing effect has not been investigated previously, we here show that in the presence of soybean 15LO (SLO) or human 15LO (rhLO), the addition of lipoprotein lipase, porcine pancreatic, or human type IIa secretory phospholipase A2 (sPLA2) greatly enhanced the accumulation of hydro(pero)xides of all major classes of LDL's lipids. Hydroperoxides of free fatty acids accumulated exclusively as enzymic products with kinetics reflecting both the formation of free fatty acids and the initial 'build-up' of alpha-tocopheroxyl radical. In contrast, hydroperoxides of cholesteryl esters and phosphatidylcholine accumulated linearly over comparatively longer periods of time and, in the case of rhLO, well beyond inactivation of the oxygenase. With SLO, formation of oxidized esterified lipids occurred nonenzymically, independent of the presence of lipase and despite the oxygenase remaining active until the end of the incubation. Enhancement of rhLO-induced LDL lipid peroxidation by sPLA2 was eliminated by a neutralizing anti-sPLA2 antibody, indicating that lipolytic activity was required for this effect. LDL depleted of alpha-tocopherol was resistant to oxidation by 15LO alone, whereas lipase overcame this resistance, demonstrating that lipases enhance 15LO-induced enzymic and nonenzymic peroxidation of LDL lipids. This is likely due to provision of free fatty acid substrate, resulting in an enhanced rate of free radical formation which itself causes nonenzymic peroxidation of esterified lipids. As lipases and 15LO are present in atherosclerotic lesions, our findings could be of pathophysiological significance.

Animals↗

A stable-isotope approach to delineate geographical catchment areas of avian migration monitoring stations in North America.

Migration monitoring stations (MMSs) were established to provide information on population trends of North American passerines. However, apart from inferring general origins of birds, there has been no way to delineate geographical catchment areas sampled by MMSs. The ability to resolve MMS catchment areas would greatly enhance our ability to link and constrain population declines to specific geographical areas and thereby focus conservation efforts. Here, we utilize stable-hydrogen (delta D) and carbon (delta 13C) isotope values of rectrices of fall hatch-year (HY) and spring adult after-second year (ASY) Swainson's Thrush (Catharus ustulatus) moving through two MMSs in Canada to determine natal and molting catchment areas for those stations. Stable-carbon isotope signatures provided no information on geographical origins of birds. Conversely, delta D signatures provided good latitudinal geographical control, and this was also supported by analysis of feathers from two other species, Gray-cheeked Thrush (Catharus minimus) and Harris' Sparrow (Zonotrichia querula), also known to breed and molt in northern regions. The delta D values of fall HY birds moving through Delta Marsh MMSs in Manitoba were more negative than those moving through Long Point MMSs in southern Ontario, clearly indicating more northwesterly origins. However, based on delta D values alone, both MMSs sampled birds from broad catchment regions of the Canadian Boreal forest. The interpolated catchment area of the Delta MMS encompassed northwest Manitoba to northwestern Alberta. The interpolated catchment area of the Long Point MMS extended from north central Ontario and Quebec and into western Canada. The delta D values of birds moving northward in spring suggested that most ASY Swainson's Thrush molted at more southerly locations than their breeding sites. We show the stable-hydrogen isotope approach may be employed to significantly improve conventional observational techniques for avian population monitoring, and that MMSs provide a reliable means of associating population productivity with regional conservation issues.

Animals↗

Improved method for quantifying the avicide 3-chloro-p-toluidine hydrochloride in bird tissues using a deuterated surrogate/GC/MS method.

A method using a deuterated surrogate of the avicide 3-chloro-p-toluidine hydrochloride (CPTH) was developed to quantify the CPTH residues in the gastrointestinal (GI) tract and breast muscle tissues in birds collected in CPTH-baited sunflower and rice fields. This method increased the range of a previous surrogate/gas chromatography/mass spectroscopy method from 0-2 to 0-20 microg/g in tissue samples and greatly simplified the extraction procedure. The modified method also sought to increase recoveries over a range of matrix effects introduced by analyzing tissues from birds collected in the field, where the GI tract contents would be affected by varying diet. The new method was used to determine the CPTH concentration in GI tract samples fortified with CPTH-treated rice bait to simulate the consumption of varying amounts of treated bait by two nontargeted bird species, pigeon (Columbia livia) and house sparrow (Passer domesticus). The new method was then used to examine the CPTH concentrations in the gizzard contents of the targeted bird species, red-winged black bird (Agelaius phoeniceus) and brown-headed cowbird (Molothrus ater), that were collected after feeding at a treated bait site. The method proved sufficiently sensitive to quantify CPTH in the breast muscle tissues and the gizzard contents of red-winged blackbirds and brown-headed cowbirds during an operational baiting program. The levels of CPTH determined for these birds in both tissue samples were determined to be highly correlated. The appearance of CPTH in the breast muscle tissue immediately after feeding was not anticipated. The potential secondary hazard posed by the targeted birds to potential scavengers and predators was also evaluated.

Animals↗