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Sexual dimorphism of vocal control nuclei in budgerigars (Melopsittacus undulatus) revealed with Nissl and NADPH-d staining.

Nissl staining and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry were used to explore the existence of sexual dimorphism in vocal control nuclei of adult budgerigars (Melopsittacus undulatus), a parrot species capable of lifelong vocal learning. Behavioral studies indicate that adult males possess larger vocal repertoires than adult females and learn new calls more quickly. The results of the present study show that the volumes of all vocal nuclei, as measured using both Nissl-stained and NADPH-d-stained material, as well as the total numbers of NADPH-d neurons, were 35-110% greater in males. Furthermore, all vocal nuclei exhibit conspicuous NADPH-d staining compared to surrounding fields in both adult males and females. Nevertheless, there were no significant gender differences in either the intensity of neuropil staining or the densities of NADPH-d neurons in vocal nuclei. Moreover NADPH-d neuron somal shapes were similar in males and females. Diameters of NADPH-d neurons in vocal nuclei were 8.5-32% larger in males than in females. Greater size of NADPH-d neuronal somata in males may be a general property of this cell type in budgerigars because a similar gender difference was found in a visual nucleus, the entopallium, which is not directly associated with the vocal control system and does not exhibit sexual dimorphism in total volume or total NADPH-d neuron numbers. Taken together, the results of the present study favor the hypothesis that superior lifelong vocal learning ability in male budgerigars rests largely on larger volumes of vocal control nuclei in males rather than on sexual dimorphism in the internal composition of vocal nuclei.

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Color vision of the budgerigar (Melopsittacus undulatus): hue matches, tetrachromacy, and intensity discrimination.

Budgerigars, Melopsittacus undulatus, were trained to discriminate monochromatic lights from mixtures of two comparison lights. The addition of small amounts of UV (365 nm) to blue or yellow lights dramatically changed the color for the birds. Hue matches showed the birds to be dichromatic both at long wavelengths (only P565 and P508 active) and at short wavelengths (only P370 and P445 active because of screening of P508 and P565 by cone oil droplets). In mid-spectrum (only P445 and P508 active), a hue match was achieved, but the results were more complicated because two opponent neural processes were activated. All observed hue matches were in quantitative agreement with calculations of relative quantum catch in the pairs of participating single cones and point to the presence of a minimum of three opponent neural processes. For the hue matches at mid- and short wavelengths, the calculations also predict peak values of absorbance of the cone oil droplets associated with P508 and P445. Relative intensity of the training light affected difficult matches at long but not short wavelengths, likely due to achromatic signals from the double cones. With suitable training, birds could make intensity discriminations at short wavelengths, where the double cones have diminished sensitivity.

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Multidrug-resistant bacterial ingluvitis associated with squamous cell carcinoma in a budgerigar (Melopsittacus undulatus).

A 2-year-old budgerigar (Melopsittacus undulatus) was presented to the Massey University Veterinary Teaching Hospital for chronic regurgitation. Multiple drug-resistant Klebsiella oxytoca was cultured from the crop. Necropsy revealed a mass in the crop that was histologically diagnosed as squamous cell carcinoma (SCC). To the authors' knowledge, this is the first report of SCC in a budgerigar associated with a multidrug-resistant bacterial ingluvitis.

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The precedence effect in three species of birds (Melopsittacus undulatus, Serinus canaria, and Taeniopygia guttata).

The perceived locations of paired auditory images, simulating direct sounds and their echoes, have been recently studied in budgerigars (Melopsittacus undulatus; M. L. Dent & R. J. Dooling, 2003a, 2003b). In this article, the authors extend those experiments to include measurements of the precedence effect using a discrimination paradigm in two additional bird species: canaries (Serinus canaria) and zebra finches (Taeniopygia guttata). Although time courses of summing localization, localization dominance, and echo thresholds were similar across all species, budgerigars had slightly higher overall levels of discrimination. The results from these experiments add further support that the precedence effect in birds is similar to that found in other animals and that the ability to suppress echoes that might degrade localization and auditory object perception may be a general property of the vertebrate auditory system.

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Comparitive sensitivity of polymerase chain reaction diagnosis of psittacine beak and feather disease on feather samples, cloacal swabs and blood from budgerigars (Melopsittacus undulates, Shaw 18005).

A longitudinal study was performed in order to investigate virus excretion and viraemia during a clinical outbreak of the psittacine beak and feather disease in budgerigars (Melopsittacus undulatus). Viral nucleic acid was detected in feathers, cloacal swabs and blood samples. Overall, beak and feather disease virus (BFDV) DNA was detected most commonly in feather samples, followed by cloacal swabs, and least frequently from blood samples. In most cases the viraemia was short lived and correlated with clinical signs, such as feather abnormalities. Sequence analysis of the polymerase chain reaction fragment amplified from the replication-associated gene (ORF V1) indicated a close relationship with other BFDV isolates. Overall the highest level of nucleotide identity was found with the ORF V1 of another budgerigar isolate. Our results suggest that feather samples and cloacal swabs should be taken for polymerase chain reaction diagnosis to determine the presence of BFDV in an aviary, but that detection in these samples may not correlate well with psittacine beak and feather disease.

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Retrobulbar rhabdomyosarcoma in a budgerigar (Melopsittacus undulatus).

This report describes a retrobulbar rhabdomyosarcoma in a 7-year-old male budgerigar (Melopsittacus undulatus). The bird was presented with orbital distension and exophthalmia of the left eye and died during surgery. Necropsy revealed a tan to grey retrobulbar mass compressing all components of the left eye with obscured vision. In histopathological examination, the mass was composed of sheets of spindle-shaped cells with pleomorphic nuclei, numerous bizarre mitotic figures and mononucleated or multinucleated giant cells and also typically strap cells. Neoplastic cells had no cross-striations in sections stained by phosphotungstic acid haematoxylin. The tumour cells did not invade surrounding tissues, including the retina, and there was no metastasis to other organs. Immunohistochemically, tumour cells were positive for desmin, vimentin and alpha-smooth muscle actin, but were negative for S-100 protein and pancytokeratin.

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Contact-call driven and tone-driven zenk expression in the nucleus ovoidalis of the budgerigar (Melopsittacus undulatus).

The effectiveness of species-typical contact calls and a 3-kHz pure tone to induce zenk gene protein expression in the primary thalamic auditory relay nucleus ovoidalis was compared in budgerigars (Melopsittacus undulatus), a parrot species capable of lifelong vocal learning. Ovoidalis consists of a core which projects topographically to field L of the telencephalon and a ventromedial shell containing many calcitonin-gene-related peptide neurons that project throughout field L as well as to an adjacent field receiving visual input. Tone-induced and call-induced zenk expression in the ovoidalis core were similar; however, call-induced zenk expression in ventromedial ovoidalis shell was significantly greater than tone-induced expression. These results support the idea that the ovoidalis shell may contain neurons specialized to process complex sounds including species-typical communication sounds.

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Clinical approach to renal neoplasia in budgerigars (Melopsittacus undulatus).

Renal neoplasia in budgerigars (Melopsittacus undulatus) is seen regularly in general practice and is of interest to many practicing veterinary surgeons. This review article provides an overview of the current knowledge and the most recent reports in avian literature regarding renal tumours in budgerigars, with the emphasis on clinical diagnosis and treatment. The high prevalence of renal neoplasia in budgerigar is discussed, with notes on the most commonly diagnosed tumours, possibility of metastases, sex and age predisposition. The possibility of ultrasonography and radiography in the diagnosis of this pathological condition are compared as well as the possible role of blood and urine analyses. Two studies are described, both of which investigate the possible involvement of a retrovirus as the aetiological agent.

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Effects of two diets on the haematology, plasma chemistry and intestinal flora of budgerigars (Melopsittacus undulatus).

Two groups of 22 budgerigars (Melopsittacus undulatus) were housed for 12 months under identical conditions. One group was fed a commercial seed mixture plus carrots and a mineral supplement, and the other group was fed a commercially formulated diet plus carrots. Samples of blood and faeces were collected initially and after three, six, nine and 12 months. There were no significant differences between the haematological values of the two groups. The group fed the seed mixture had significantly higher concentrations of glucose, albumin, triglycerides and uric acid, and higher activity of aspartate aminotransferase, but the values were within the published reference ranges for normal birds. There were no significant differences between the faecal samples from the two groups, except that the fungus Macrorhabdus ornithogaster was identified in 48.3 per cent of the samples from the group fed the commercially formulated diet but from only 3.4 per cent of the samples from the group fed the seed mixture.

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The vasotocinergic system in the hypothalamus and limbic region of the budgerigar (Melopsittacus undulatus).

We report a morphological and biochemical analysis on the presence, distribution and quantification of vasotocin in the hypothalamus and limbic region of the budgerigar Melopsittacus undulatus, using immunohistochemistry on serial sections and competitive enzyme linked immunoadsorbent assay measurements on tissue extracts. Analysis of the sections showed large vasotocin-immunoreactive neurons in three main regions of the diencephalon, of both male and female specimens. Vasotocinergic cell bodies were located in the ventral and lateral areas of the hypothalamus, dorsal to the lateral thalamus and medial to the nucleus geniculatus lateralis. Immunoreactive neurons were placed also periventricularly, close to the walls of the third ventricle, at the level of the magnocellular paraventricular nucleus. Well evident bundles of immunoreactive fibers were placed ventral to the anterior commissure in the same regions of the hypothalamus and thalamus where vasotocinergic perikarya are localized. Fibers were identified close to the third ventricle, and in the lateral hypothalamic area along the lateral forebrain bundle. In contrast to what reported for other oscine and non-oscine avian species, we were not able to identify immunopositive neurons in any region above the anterior commissure, or detect relevant differences on the distribution of the vasotocin immmunoreactivity between sexes. Competitive enzyme linked immunoadsorption assay and image analysis of the extension of immunoreactivity in the tissue sections were consistent with the qualitative observations and indicated that there is no statistically significant dimorphism in the content of vasotocin or in the location and distribution of vasotocinergic elements in the investigated areas of male and female parrot brains.

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Biological variation, individuality and critical differences of eight biochemical blood constituents in budgerigars (Melopsittacus undulatus).

The aim of the study was to obtain basic data on individual biological variation, the required number of specimens to define the homeostatic setpoint (the aspired value of a variable adjusted to the individuals homeostasis) and critical differences of selected chemistry parameters in budgerigars (Melopsittacus undulatus). Blood from 99 healthy budgerigars was sampled 12 times at four-week intervals. Aspartate aminotransferase (ast), calcium, creatine kinase (ck), glutamate dehydrogenase (gldh), glucose, lactate dehydrogenase (ldh), total protein and uric acid were investigated. The indices of individuality obtained in the present study were relatively low (total protein 0.93, ast 1.02, gldh 1.04, ck 1.12, ldh 1.24, uric acid 1.26, glucose 1.39, calcium 1.61) and suggest that population-based reference limits might be useful. Comparison of data showed that the application of intraindividual reference values identified much greater variation in the reference values than using conventional population-based reference limits. Otherwise, the moderate to low degrees of individuality may allow the use of reference values of one individual as a standard for another individual of the same species. The critical differences that define the change needed between two serial results to indicate a biological change was highest in gldh with 120 per cent or 1.7 U/l. In calcium, a change of 30.5 per cent or 0.5 mmol/l would be significant. In the other parameters critical differences varied between 38 per cent in glucose, up to 93 per cent in uric acid.

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Distribution of choline acetyltransferase and acetylcholinesterase in vocal control nuclei of the budgerigar (Melopsittacus undulatus).

The present study used histochemical methods to map the distributions of choline acetyl transferase (ChAT) and acetylcholinesterase (AChE) in the vocal control nuclei of a psittacine, the budgerigar (Melopsittacus undulatus). The distributions of ChAT and AChE in budgerigars appeared similar to that in oscine songbirds despite evidence that these systems have evolved independently. The magnicellular nucleus of the lobus parolfactorius in budgerigars, like the area X in songbirds, contained many ChAT labeled somata, fibers, and varicosities and stained densely for AChE. In contrast, the robust nucleus of the archistriatum (RA) and the supralaminar area of the frontal neostriatum in budgerigars, like the RA and the magnicellular nucleus of the neostriatum (MAN) in songbirds, respectively, contained few or no ChAT labeled somata, fibers, and varicosities and stained lightly for AChE. The central nucleus of the lateral neostriatum in budgerigars, like the higher vocal center (HVC) in songbirds, contained no ChAT labeled somata, moderate densities of ChAT labeled fibers and varicosities, and moderate levels of AChE staining. Two nuclei, the oval nucleus of the hyperstriatum ventrale (HVo) and the oval nucleus of the anterior neostriatum (NAo), contained no ChAT labeled somata, dense ChAT labeled fibers and varicosities, and moderate to high levels of AChE staining. The HVo and the NAo have no counterparts in songbirds but may be important vocal control nuclei in the budgerigar. Cholinergic enzymes are also described in other regions which may be involved in budgerigar vocal behavior, including the basal forebrain, the torus semicircularis, and the hypoglossal nuclei (nXII).

Acetylcholinesterase↗

Vocal control pathways through the anterior forebrain of a parrot (Melopsittacus undulatus).

A feature of the telencephalic vocal control system in the budgerigar (Melopsittacus undulatus) that has been hypothesized to represent a profound difference in organization from the oscine vocal system is its reported lack of an inherent circuit through the anterior forebrain. The present study reports anatomical connections that indicate the existence of an anterior forebrain circuit comparable in important ways to the "recursive" pathway of oscine songbirds. Results from anterograde and retrograde tracing experiments with biocytin and fluorescently labeled dextran amines indicate that the central nucleus of the anterior archistriatum (AAc) is the source of ascending projections upon the oval nuclei of the anterior neostriatum and ventral hyperstriatum (NAo and HVo, respectively). Efferent projections from the latter nuclei terminate in the lateral neostriatum afferent to AAc, thereby forming a short recurrent pathway through the pallium. Previously reported projections from HVo and NAo upon the magnocellular nucleus of the lobus parolfactorius (LPOm), and after LPOm onto the magnocellular nucleus of the dorsal thalamus (DMm; G.F. Striedter [1994] J. Comp. Neurol. 343:35-56), are confirmed. A specific projection from DMm onto NAom is also demonstrated; therefore, a recurrent pathway through the basal forebrain also exists in the budgerigar vocal system that is similar to the anterior forebrain circuit of oscine songbirds. Parallels between these circuits and mammalian basal ganglia-thalamo-cortical circuits are discussed. It is hypothesized that vocal control nuclei of the avian anterior neostriatum may perform a function similar to the primate supplemental motor area.

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Methionine enkephalin immunoreactivity in the brain of the budgerigar (Melopsittacus undulatus): similarities and differences with respect to oscine songbirds.

The brain of the budgerigar (Melopsittacus undulatus), a small parrot that acquires new vocalizations throughout life, was examined for immunoreactivity to the opioid peptide methionine enkephalin (mENK). mENK is a highly prominent feature of the chemical architecture of the forebrain vocal system of oscine songbirds. Forebrain vocal control nuclei are believed to have evolved independently in parrots and songbirds (Streidter [1994] J. Comp. Neurol. 343:35-56); however, recent studies have found similarities in the neural organization of vocal control pathways in budgerigars and songbirds (Durand et al. [1997] J. Comp. Neurol. 377:179-206). Among the similarities are the existence of recursive pathways interconnecting vocal control neurons in the archistriatum, basal ganglia (i.e., lobus parolfactorius), and dorsal thalamus. In the present study, we found that all vocal control nuclei within the budgerigar forebrain exhibit prominent mENK-like immunoreactivity (ELI) in fibers and somata. We also found striking similarities between the morphology of ELI elements in budgerigar vocal control nuclei and that described previously in songbird vocal nuclei. Despite these similarities, the budgerigar dorsal striatopallidum (lobus parolfactorius, paleostriatum augmentatum, and paleostriatum primitivum) and somatomotor (anterior) archistriatum exhibit unique patterns of ELI. The dorsal striatopallidum contained far less ELI, whereas the archistriatum contained far more than would be expected on the basis of previous studies of opioid peptides in other avian species, including pigeons, chickens, and songbirds. These differences may reflect neural specializations unique to the budgerigar that contribute to the extraordinary flexibility of the vocal motor system of this species to acquire socially significant stimuli throughout life.

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Distribution of tyrosine hydroxylase-containing neurons and fibers in the brain of the budgerigar (Melopsittacus undulatus): general patterns and labeling in vocal control nuclei.

The distribution of tyrosine hydroxylase (TH) was mapped out in cells and fibers of the budgerigar (Melopsittacus undulatus) brain. Special attention was given to vocal control and auditory nuclei because budgerigars are a psittacine species in which both males and females are capable of lifelong vocal learning (Farabaugh et al. [1994] J. Comp. Psychol 108:81-92). The results show that TH staining in the central nucleus of the anterior archistriatum (AAc) resembled that of surrounding archistriatal fields, except for portions of the ventral archistriatum, which exhibited substantially more TH+ fibers. Fewer fibers and fiber baskets are present in the central nucleus of the lateral neostriatum (NLc) than in surrounding fields. Both the oval nuclei of the ventral hyperstriatum (HVo) and anterior neostriatum (NAo) exhibit less fiber staining than surrounding fields whereas fiber staining in the medial NAo (NAom) and magnicellular nucleus of the parolfactory lobe (LPOm) resemble that of surrounding fields. Staining in primary telencephalic auditory nuclei was extremely low. The only sex difference observed was slightly increased TH staining in LPOm of females compared with surrounding fields on some tissue sections. These findings are in contrast to previous findings in zebra finch (Poephila guttata), a close ended vocal learning songbird in which TH staining in vocal nuclei increases during development and remains greater than surrounding fields throughout adulthood. The present results therefore support the view that catecholamines act to inhibit vocal plasticity in adult vocal learning species. Several unique features of TH-immunoreactive (ir) cell groups were observed in the brainstem including sparsely scattered TH-ir somata immediately adjacent to the third ventricle, within the tectum, basal forebrain, archistriatum, and caudal neostriatum, and in the hippocampus. These latter populations have not been described in other avian species and resemble features of the catecholamine system generally found in either reptiles or mammals.

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Microscopic and submicroscopic anatomy of the parabronchi, air sacs, and respiratory space of the budgerigar (Melopsittacus undulatus).

The normal microscopic and submicroscopic structure of the lower respiratory tract of the budgerigar (Melopsittacus undulatus) is described and compared with other birds and mammals. Granular (type II) pneumocytes are confined to linings of air sacs, parabronchi, and their atria; however, their secretions (surfactant) cover the surfaces of the infundibula and respiratory space. Infundibula extend from the atria and give rise to the air capillaries, which branch and anastomose freely with those of adjacent infundibula and other parabronchi (interparabronchial septa are not found). Infundibula and the respiratory labyrinth are lined by a continuous epithelium of squamous pneumocytes, whose perikarya are concentrated in the infundibula and whose peripheral cytoplasm is markedly attenuated. The squamous pneumocytes of the respiratory labyrinth share a basal lamina with the blood capillaries that they envelop.

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Organization of the avian basal forebrain: chemical anatomy in the parrot (Melopsittacus undulatus).

Hodological, electrophysiological, and ablation studies indicate a role for the basal forebrain in telencephalic vocal control; however, to date the organization of the basal forebrain has not been extensively studied in any nonmammal or nonhuman vocal learning species. To this end the chemical anatomy of the avian basal forebrain was investigated in a vocal learning parrot, the budgerigar (Melopsittacus undulatus). Immunological and histological stains, including choline acetyltransferase, acetylcholinesterase, tyrosine hydroxylase, dopamine and cAMP-regulated phosphoprotein (DARPP)-32, the calcium binding proteins calbindin D-28k and parvalbumin, calcitonin gene-related peptide, iron, substance P, methionine enkephalin, nicotinamide adenine dinucleotide phosphotase diaphorase, and arginine vasotocin were used in the present study. We conclude that the ventral paleostriatum (cf. Kitt and Brauth [1981] Neuroscience 6:1551-1566) and adjacent archistriatal regions can be subdivided into several distinct subareas that are chemically comparable to mammalian basal forebrain structures. The nucleus accumbens is histochemically separable into core and shell regions. The nucleus taeniae (TN) is theorized to be homologous to the medial amygdaloid nucleus. The archistriatum pars ventrolateralis (Avl; comparable to the pigeon archistriatum pars dorsalis) is theorized to be a possible homologue of the central amygdaloid nucleus. The TN and Avl are histochemically continuous with the medial aspects of the bed nucleus of the stria terminalis and the ventromedial striatum, forming an avian analogue of the extended amygdala. The apparent counterpart in budgerigars of the mammalian nucleus basalis of Meynert consists of a field of cholinergic neurons spanning the basal forebrain. The budgerigar septal region is theorized to be homologous as a field to the mammalian septum. Our results are discussed with regard to both the evolution of the basal forebrain and its role in vocal learning processes.

Acetylcholinesterase↗

Relation of medullary motor nuclei to nerves supplying the vocal tract of the budgerigar (Melopsittacus undulatus).

The locations of brainstem motor nuclei supplying efferents to organs of the vocal tract of the budgerigar (Melopsittacus undulatus) were determined as a first step in describing brain pathways for vocal control in this species. We identified four major nerves supplying the muscles of the syrinx, larynx, and tongue, and separately treated each of these with horseradish peroxidase. Nerves supplying efferents to the larynx and tongue arise from ipsilateral cells in the nucleus ambiguus and hypoglossal nucleus, respectively. Each right and left half of the syrinx is supplied by both right and left halves of the hypoglossal nucleus, via axons crossing in the common anastomosis of the tracheosyringeal nerves. This bilateral innervation of the syrinx will allow testing for central asymmetries of dominance for vocal control in the absence of any underlying anatomical laterality in the periphery.

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