A habilidade para produzir e usar ferramentas é um marco na evolução humana, mas outros animais também o fazem. Na maioria das vezes esse comportamento é atribuído a uma predisposição genética e não a evidências de que entendam as relações de causa e efeito envolvidas. Observamos araras azuis (Anodorhynchus hyacinthynus) durante a manipulação e quebra de cocos (Attalea dubia, em cativeiro e Acrocomia totai, na natureza), encontrando evidências a uma tendência inata ao uso de ferramentas. Diferenças de técnica e proficiência entre jovens e adultos sugerem que o aperfeiçoamento exige aprendizagem. Os cocos são selecionados e descartados com algum critério pelas aves adultas. Aves selvagens usam apenas uma ferramenta, momentos antes da ruptura da cápsula, sugerindo que reduza o impacto. Aparentemente as ferramentas não diminuem o tempo de quebra, mas reduzem o número de cocos descartados. Todas as ferramentas materiais foram usadas do mesmo modo e apenas para a quebra de cocos, não havendo correlação entre o material, ordem e tempo de uso. O teste do String Pulling revelou que as araras azuis e outras espécies de psitacídeos são capazes de identificar relações de causa-e-efeito entre objetos relevantes para a resolução de problemas / The ability to produce and use tools is a milestone in human evolution, but other animals also do it. Most often this behavior is attributed to a genetic predisposition rather than to evidence that they understand cause-and-effect relations involved. We observed hyacinth macaws (Anodorhynchus hyacinthynus) during the handling and breaking of nuts (Attalea dubia in captivity and Acrocomia totai in nature), finding evidence for an innate tendency to use tools. Differences in technique and proficiency among young and adult birds suggest that tool use improvement requires learning. The nuts are selected and discarded with some criteria by adult birds. Frequently, wild birds used only one tool, moments before nut rupture, suggesting that the tool is used for impact reduction. Apparently the tools do not reduce the time needed for breaking the nut, but they seem to reduce the number of non-eaten nuts. All materials were used in the same manner and only to the breaking of nuts. There was no correlation between material, order and duration of tool use. The String Pulling test revealed that the birds are able to identify cause-and-effect relationship, as well as relevant properties of the physical world involved in the solution of the problem

A habilidade para produzir e usar ferramentas é um marco na evolução humana, mas outros animais também o fazem. Na maioria das vezes esse comportamento é atribuído a uma predisposição genética e não a evidências de que entendam as relações de causa e efeito envolvidas. Observamos araras azuis (Anodorhynchus hyacinthynus) durante a manipulação e quebra de cocos (Attalea dubia, em cativeiro e Acrocomia totai, na natureza), encontrando evidências a uma tendência inata ao uso de ferramentas. Diferenças de técnica e proficiência entre jovens e adultos sugerem que o aperfeiçoamento exige aprendizagem. Os cocos são selecionados e descartados com algum critério pelas aves adultas. Aves selvagens usam apenas uma ferramenta, momentos antes da ruptura da cápsula, sugerindo que reduza o impacto. Aparentemente as ferramentas não diminuem o tempo de quebra, mas reduzem o número de cocos descartados. Todas as ferramentas materiais foram usadas do mesmo modo e apenas para a quebra de cocos, não havendo correlação entre o material, ordem e tempo de uso. O teste do String Pulling revelou que as araras azuis e outras espécies de psitacídeos são capazes de identificar relações de causa-e-efeito entre objetos relevantes para a resolução de problemas / The ability to produce and use tools is a milestone in human evolution, but other animals also do it. Most often this behavior is attributed to a genetic predisposition rather than to evidence that they understand cause-and-effect relations involved. We observed hyacinth macaws (Anodorhynchus hyacinthynus) during the handling and breaking of nuts (Attalea dubia in captivity and Acrocomia totai in nature), finding evidence for an innate tendency to use tools. Differences in technique and proficiency among young and adult birds suggest that tool use improvement requires learning. The nuts are selected and discarded with some criteria by adult birds. Frequently, wild birds used only one tool, moments before nut rupture, suggesting that the tool is used for impact reduction. Apparently the tools do not reduce the time needed for breaking the nut, but they seem to reduce the number of non-eaten nuts. All materials were used in the same manner and only to the breaking of nuts. There was no correlation between material, order and duration of tool use. The String Pulling test revealed that the birds are able to identify cause-and-effect relationship, as well as relevant properties of the physical world involved in the solution of the problem

Submitted by Elís Domingos Ferrari null (elisd.ferrari@yahoo.com.br) on 2017-10-31T11:41:02Z No. of bitstreams: 1 FINAL-Defesa Mestrado CORRIGIDO.pdf: 1177720 bytes, checksum: 7a08f0cef1e5a124653c8190f8178822 (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-11-10T18:35:51Z (GMT) No. of bitstreams: 1 ferrari_ed_me_araca_par.pdf: 606404 bytes, checksum: 87114f79e3b3cffb7d3ab08cd9009eb6 (MD5) / Made available in DSpace on 2017-11-10T18:35:51Z (GMT). No. of bitstreams: 1 ferrari_ed_me_araca_par.pdf: 606404 bytes, checksum: 87114f79e3b3cffb7d3ab08cd9009eb6 (MD5) Previous issue date: 2017-10-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O objetivo deste estudo foi avaliar a ocorrência e os métodos de diagnóstico para Cryptosporidium spp. em psitacídeos exóticos de cativeiro provenientes das regiões sul e sudeste do Brasil. A purificação dos oocistos nas amostras fecais de 463 psitacídeos foi realizada por meio de centrifugo-flutuação em solução de Sheather. Para análise microscópica, nós utilizamos a coloração negativa de verde malaquita. A amplificação de um fragmento parcial do gene 18S rRNA de Cryptosporidium spp. foi feita usando-se nested PCR seguida de sequenciamento dos fragmentos amplificados (nPCR/S). As amostras também foram testadas por meio de PCR duplex em tempo real, visando-se amplificar um fragmento do gene 18S rRNA de Cryptosporidium galli e Cryptosporidium genótipo III de aves. A ocorrência de Cryptosporidium spp. pela microscopia e nested PCR (nPCR) foi de 3, 02% (14/463) e 4, 97% (23/463), respectivamente. A nPCR/S demonstrou positividade de 1, 73% (8/463) para Cryptosporidium genótipo III de aves, 0, 86% (4/463) para Cryptosporidium parvum e 0, 22% (1/463) para Cryptosporidium canis. A PCR duplex em tempo real demonstrou positividade de 9, 50% (44/463) para as criptosporidiose gástrica, sendo 1, 94% (9/463) para C. galli, 5, 83% (27/463) para Cryptosporidium genótipo III de aves e 1, 73% (8/463) para infecções mistas. Não houve diferença estatística significante entre a positividade pela nPCR e microscopia (p = 0. 1237) e houve concordância justa entre elas (Kappa = 0. 242). Diferença estatística significante (p <0. 0001) e concordância justa (Kappa = 0. 317) foram obtidas nas comparações entre nPCR e PCR duplex em tempo real. Nós concluímos que a PCR duplex em tempo real é a melhor opção para o diagnóstico de criptosporidiose gástrica e que Cryptosporidium genótipo III de aves é o mais comum dentre as espécies/ genótipos de Cryptosporidium que acometem psitacídeos. / The aim of this study was to evaluate the prevalence and diagnostic methods for Cryptosporidium spp. in caged adult exotic parrots from Southern and Southeastern Brazil. The oocyst purification in fecal samples from 463 psittacines was performed by centrifugal-flotation in Sheather's sugar solution. For microscopic analysis, we used malachite green negative staining. Amplification of a partial fragment of the 18S rRNA gene of Cryptosporidium spp. was accomplished using nested PCR (nPCR) followed by sequencing of the amplified fragments (nPCR/S). Samples were also tested by duplex real-time PCR targeting the 18S rRNA gene of Cryptosporidium galli and Cryptosporidium avian genotype III. The prevalence rates of Cryptosporidium spp. by microscopy and nPCR was 3. 02% (14/463) and 4. 97% (23/463), respectively. The nPCR/S showed positivity of 1. 73% (8/463) for Cryptosporidium avian genotype III, 0. 86% (4/463) for Cryptosporidium parvum and 0. 22% (1/463) for Cryptosporidium canis. Duplex real-time PCR showed a positivity of 9. 50% (44/463) for gastric cryptosporidiosis, 1. 94% (9/463) for C. galli, 5. 83% (27/463) for Cryptosporidium avian genotype III and 1. 73% (8/463) for mixed infections. There was no statistically significant difference between positivity for nPCR and microscopy (p = 0. 1237) and fair agreement between them (Kappa = 0. 242). A significant statistical difference (p <0. 0001) and fair agreement (Kappa = 0. 317) were obtained between nPCR and duplex real-time PCR. We found out that duplex real-time PCR is the best option for the diagnosis of gastric cryptosporidiosis and that Cryptosporidium avian genotype III is the most common Cryptosporidium species /genotype in psittacines. / FAPESP: 2015/26334-8

Das Ziel dieser Arbeit war es, das Herz von gesunden Psittaciformes und insbesondere des Graupapageis mithilfe der Ultraschall-Technik zu untersuchen. Die darstellbaren Strukturen sollten vermessen und – soweit möglich - anatomische sowie physiologische Referenzwerte aufgestellt werden, um eine Grundlage für die Beurteilung von Herzen bei Psittaciformes mit Anzeichen einer Herz-Kreislauf-Erkrankung zu erhalten. Außerdem sollte die Darstellbarkeit des Herzens im Ultraschallbild zwischen verschiedenen aus unterschiedlichen geographischen Regionen stammenden Psittaciformes verglichen werden. Zu diesem Zweck wurden 110 Psittaciformes untersucht. Alle Vögel wurden zunächst einer ausführlichen Gesundheitskontrolle unterzogen, um Tiere mit Hinweis auf eine Störung im Herz-Kreislauf-System von der Studie ausschließen zu können. Desweiteren wurden verschiedene Größenparameter vermessen (Brustbeinlänge, Bauchlänge, Körpermasse), um die erhaltenen Ultraschall-Meßwerte in Relation zur Tiergröße zu vergleichen. Die Vögel wurden für die Untersuchung auf eine eigens konstruierte Fixationseinrichtung verbracht, und mit ebenfalls speziell für die Untersuchung entwickelten Elektroden wurde das Elektrokardiogramm abgeleitet. Dieses diente zur Triggerung von Endsystole und Enddiastole im Ultraschallbild. Die Untersuchung erfolgte mit einem elektronischen 6,5-MHz-Micro-Curved-Array-Schallkopf (betrieben auf 7,5 MHz) über eine Ankopplung in der Medianen unmittelbar hinter dem Brustbein; durch Rotation des Schallkopfes um 90° entstanden ein vertikaler und ein horizontaler Schnitt durch das Herz. Hinsichtlich der Darstellbarkeit des Herzens im Ultraschallbild zeigten sich deutliche Unterschiede zwischen den untersuchten Tiergruppen. Die Größe der Tiere spielte insofern eine Rolle, als daß bei den kleineren Vögeln eine entsprechende Ankopplung des verwendeten Schallkopfes schwierig war, deshalb konnte bei Psittaciformes in der Größe des Wellen- bzw. Mönchssittichs das Herz nicht dargestellt werden. Unabhängig von der Größe der Tiere zeigte sich aber auch eine speziesabhängige Schallbarkeit. Generell ließ sich bei den untersuchten Graupapageien und Amazonen das Herz gut darstellen, bei den Kakadus war die Darstellungsqualität dagegen wesentlich schlechter. Bei den untersuchten Tieren konnten abhängig von der Darstellungsqualität verschiedene Herzdimensionen vermessen werden. So ließen sich die Herzkammern sowie der linke Vorhof in der Länge und der Breite vermessen, außerdem das Interventrikularseptum und die Aorta. Da die Messungen sowohl in der Systole als auch in der Diastole durchgeführt wurden, konnten anhand der erhaltenen Werte verschiedene Parameter der Herzarbeit errechnet werden. Für mehrere wichtige Messungen konnten erstmals Referenzwerte aufgestellt werden, so z. B. für die Abmessungen der linken Kammer, den Aortendurchmesser und die Septum-Dicke. Viele der errechneten relativen Parameter wie z. B. die Verkürzungsfraktionen, die Relation zwischen der Breite und der Länge der Kammer sowie die Ejektionsfraktion der linken Kammer zeigten konstante körpergrößenunabhängige Werte bei allen untersuchten Tiergruppen. Für einen wichtigen Parameter der Herzgröße, nämlich die Länge der linken Herzkammer, wurde eine starke Abhängigkeit von der Brustbeinlänge und der Körpermasse gefunden. Bedingt dadurch und die speziesübergreifenden relativen Parameter können auch bei Psittaciformes, bei denen noch keine Daten über echokardiographische Untersuchungen vorliegen, die physiologische Herzgröße abgeschätzt und die Herzarbeit beurteilt werden. / The aim of the study was the echocardiographical examination of the heart in healthy psittacine birds, especially in African Grey parrots. Visualizable structures should be measured and – as far as possible – anatomical as well as physiological reference values should be established in order to get a basic knowledge for the evaluation of the heart in psittacines with signs of cardiovascular disease. Furthermore, the ultrasonographical presentability of the heart and its structures should be compared between various psittacines of different continents. For this purpose 110 psittaciformes were examined. All birds were examined clinically to exclude any bird from the study with signs of a disorder in the circulatory system. Additionally several parameters for the size of the birds were measured (sternal length, abdominal length, body mass) in order to set the ultrasonographical values obtained in relation to the animal size. For the examination, the birds were restrained on a specially constructed holding device; the electrocardiogram was recorded with likewise specially developed electrodes. The electrocardiogram served for triggering the endsystolic and the enddiastolic ultrasonographical image. The examination was done with an electronical 6.5 MHz micro-curved-array transducer (working frequency 7.5 MHz) which was coupled in the median directly behind the sternum. By turning the transducer about 90° a vertical and a horizontal view through the heart was obtained. Regarding the ultrasonographical presentability of the heart, clear differences between the examined species were evident. The animal size played a part to that extent that in smaller birds the appropriate coupling of the transducer used was difficult; therefore in psittacines with a size like the budgerigars or the monk parrots the heart could not be shown. Additionally there was a difference in the ultrasonographical presentability between the species independent of the birds size. Generally the heart could be well visualized in Amazons and African Grey parrots, whereas in cockatoos the imaging quality was substantially worse. With the examined animals several heart dimensions could be measured depending on the imaging quality. The ventricles as well as the left atrium could be measured lengthwise and in the width, furthermore the interventriculary septum and the aorta. Since the measurements were carried out both in systole and diastole, different parameters of the cardiac work could be calcuted on the basis of the obtained values. It was possible to establish reference values for several important parameters for the first time, e. g. for the length and the width of the left ventricle, the aortic diameter and the thickness of the interventriculary septum. Many of the calculated relative parameters, for example the fraction shortening, the relation between the width and the length of the ventricle as well as the ejection fraction of the left ventricle showed constant values in all examined groups, which were independent on the birds size. For an important parameter of the heart size, the length of the left ventricle, a strong correlation on the sternal length and the body mass could be found. Due to this dependence and the species-spreading relative parameters it is also possible to assess the physiological heart size and the cardiac work in psittacine birds, for which no reference values exist to date.

Ziel dieser Arbeit war es, eine reproduzierbare Methode der Spermagewinnung bei Psittaciformes am Beispiel des Wellensittichs (Melopsittacus undulatus) zu entwickeln. Es sollten spermatologische Referenzwerte für diese Spezies erstellt werden, um eine erste Grundlage für deren Fruchtbarkeitsbeurteilung zu erhalten. Ferner sollten diese spermatologischen Parameter mit der endoskopischen Darstellung der Gonaden verglichen werden, um eine spezifische Beurteilungsmöglichkeit der Hodenaktivität abzuleiten. In den Vorversuchen wurden zunächst zwei verschiedene Methoden der Spermagewinnung am Wellensittich entwickelt. Die Kapillarmethode basiert auf der Entleerung der Samenspeicher (Glomus seminale) mittels einer in die Kloake eingeführten Glaskapillare. Die Massagemethode führt analog ihrer Anwendung beim Nutzgeflügel zu einer Ejakulation der Vögel. Bei beiden Techniken wurde das gewonnene Sperma in einer graduierten Glaskapillare aufgefangen. Im Vergleich der beiden verschiedenen Methoden der Spermagewinnung konnte gezeigt werden, dass hinsichtlich der höheren Spermienkonzentration und geringeren Verunreinigungen durch Erythrozyten und Urate die Massagemethode der Kapillarmethode deutlich überlegen war. Zusätzlich erwies sich die Massagemethode an versuchsfremden Wellensittichen als leicht durchführbar. Aus bereits vorliegenden Erkenntnissen über Psittaciformes konnte abgeleitet werden, dass mit der Massagetechnik innerhalb dieser Ordnung leicht Sperma gewonnen werden kann. In den verschiedenen Hauptversuchen wurde von 41 klinisch gesunden Wellensittichen aus zwei verschiedenen Zuchtformen regelmäßig mittels Massage Sperma gewonnen. Im ersten Teil wurden Proben von 19 Vermehrungszuchtwellensittichen und 10 Schauwellensittichen in zwei verschiedenen Absamintervallen spermatologisch untersucht. Die Farbe und das Volumen konnten direkt in der graduierten Glaskapillare abgelesen werden. Mittels Indikatorpapier wurde der pH-Wert bestimmt. Nach Anfärbung mit Eosin wurde für diese Ordnung erstmalig die Lebend-Tot-Rate der Spermatozoen im Ausstrich ermittelt. Im vorverdünnten Ejakulat wurde die Vorwärtsbeweglichkeit abgeschätzt. Die Dichtebestimmung wurde aus dem vorverdünnten Sperma mittels Kammerzählung bestimmt. Als Hilfswert wurde die Spermiengesamtzahl als das Produkt der Konzentration und dem Volumen berechnet. Im zweiten Hauptversuch wurden die Anteile missgebildeter Spermien nach Eosin-B-Färbung im Ejakulatausstrich von 12 weiteren Vermehrungszuchtwellensittichen bestimmt. Im letzten Versuchsteil wurde anhand der spermatologischen Ergebnisse von 37 Wellensittichen die aktuelle Hodenaktivität bestimmt und mit der endoskopischen Darstellung der Geschlechtsorgane dieser Vögel in Beziehung gesetzt. Insgesamt wiesen in den Hauptversuchsreihen sieben der 41 Vögel eine dauerhafte Azoospermie auf, ein weiterer zeigte eine hochgradige Teratozoospermie. In den erstellten Spermiogrammen des ersten Hauptversuchs wiesen beide Wellensittichrassen im 2-Tages-Absamintervall etwa die gleiche Spermiengesamtzahl auf. In der Verlängerung der Absampause von zwei auf sieben Tage stieg die Spermiengesamtzahl der ursprünglichen Vermehrungs-zuchtwellensittiche deutlicher als die der intensiv gezüchteten Schauwellensittiche. In beiden Versuchsgruppen vergrößerten sich jedoch weder das Volumen noch die Spermienkonzentration proportional zur Länge des Absamintervalls. Damit wurde offensichtlich, dass sich ein kurzes Absamintervall von 2-3 Tagen am ehesten für die Spermienanalyse und künstliche Besamung in der Ordnung der Papageienvögel eignet. Statistisch relevante Unterschiede konnten nicht für den pH-Wert und die Lebend-Tot-Rate bei den Zuchtformen nachgewiesen werden. Im zweiten Hauptversuch wurden die morphologischen Abweichungen der Spermien des Wellensittichs untersucht. Dabei dominierten Veränderungen des Spermienkopfes. Für Ziervögel wurde in den beiden Versuchen erstmalig nachgewiesen, dass die Mauser einen deutlich negativen Effekt auf die Spermaproduktion hat. Nach einem kurzen, aber deutlichen Anstieg zu Mauserbeginn erlosch die Spermienabgabe während der Mauserphase. Ebenfalls konnte in dieser Zeit eine Zunahme der Spermienmissbildungen festgestellt werden. Im Vergleich zwischen der endoskopischer Darstellung mit der Hodenaktivität konnte erstmalig nachgewiesen werden, dass die Form des Samenleiters das wichtigste Kriterium in der endoskopischen Aktivitätsbeurteilung ist, wobei der Samenleiter eines aktiven Hodens eine prallgefüllte, doppeltmäanderförmig geschlängelte Gestalt annimmt. Bestätigt werden konnte, dass aktive Hoden eine weißgraue Farbe aufweisen. Die Spermienproduktion zeigte eine signifikante Korrelation mit der Lage der kaudalen und nicht mit der kranialen Hodengrenze. Statt einer Zunahme konnte erstmalig aufgezeigt werden, dass die Gefäßzeichnung der Hoden eine geringfügige negative Korrelation mit der Aktivität aufweist. Aufgrund der anatomischen und physiologischen Verhältnisse bei anderen Ordnungen der Vögel ist möglicherweise davon auszugehen, dass sich diese Erkenntnisse artübergreifend anwenden lassen. / The aim of this study was to develop a technique of semen collection in Psittaciformes specifically in the budgerigar (Melopsittacus undulatus). In addition the investigation intended to establish sperm data as physiological reference values in order to get a basic understanding for reproductive assessment. These spermdata were then to be compared with the endoscopically determined status of the gonads, to establish a specific frame of reference for activity assessment. In the preliminary investigation two different methods of semen collection in budgerigars were developed and examined. The capillary-technique was based on an insertion of a glass capillary into the cloaca. In the course of this the semen storage (Glomus seminale) was squeezed. The massage-technique led - like in domestic-fowl - to an ejaculation of the birds. A graduated glass capillary was used to collect the released semen in both techniques. With this preliminary investigation it was shown, that semen obtained by massage technique was superior to semen obtained by capillary-technique because of the higher concentration of spermatozoa and the smaller contamination by urates and erythrocytes. In addition the massage technique was also proven on novice and untrained budgerigars. Altogether this study demonstrated that the massage technique can also be used efficiently for semen collection in the order Psittaciformes. In the main investigation the massage-stimulation was applied regularly to a total of 41 clinically healthy budgerigars of two different breeding lines. In a first phase samples were taken from 19 breeding-type budgerigars and 10 exhibition-type birds in two different collection-intervals. Colour and volume were determined directly from the glass-capillary. The pH value was measured with indicator paper. For this order for the first time the alive-dead ratio of the spermatozoa was counted via eosin staining. Forward motility was estimated in prediluted semen. In addition the sperm concentration was determined on prediluted semen via counting chamber. This lead directly to an estimate of the total sperm account. In the second part of the investigation morphologically abnormal spermatozoa of 12 further breeding-type budgerigars were classified via eosin staining and their frequency in percent was determined. In the last part of the investigation the actual testicle-activity based on these spermdata of 37 different budgerigars was compared with the endoscopically determined status of the gonads. Among the 41 budgerigars seven exhibited a permanent azoospermia. A further bird showed a constant teratozoospermia. For the two-day collection interval in the first main trial both breeding lines gave rise to approximately the same total sperm number. In extension from two to seven days collecting-interval total sperm account increased more for breeding-type than for exhibition-type birds. Besides this semen volume and sperm concentration did not increase proportionally with the extension of the interval. Thus a short collection-intervall of 2 or 3 days is most reliable for semen analysis and AI (artificial insemination) within this order. Neither pH or alive-dead ratio showed significantly differences in relation to the breeding-line and to the extension of the collection-interval. In the second part of the main investigation it was demonstrated, that malformation of the head-region dominate among morphological aberrant spermatozoa of this species. Both trials confirmed for the first time for pet-birds that moult reduces semen production significantly. Following a short increase, sperm production ceased during moulting. Additionally a significantly higher number of malformed spermatozoon’s were found. Comparing the testicle activity and status of the gonads endoscopically determined showed for the first time, that the appearance of the deferent duct is the most important criterion in endoscopic evaluation for gonadal activity. The duct of an active testicle appeared in a swollen, double-meander shape. This study also confirmed that active gonads appear whitish grey. For the endoscopic assessment of the testicular activity only the position of the caudal testicular boundary and not the cranial position was a significant criterion. With increasing sperm activity a slightly negatively correlated testicle blood vascularization could be demonstrated instead of an expected proportional increasing. Based on anatomical and physiological resemblances with other orders it is to be expected that results obtained by endoscopy may be applied to other species of birds as well.