Personality refers to multiple traits that are thought to be stable over time and across situations. It is recognized that personality has a neural basis and is associated with health outcomes. Whether personality is also associated with cognitive ability, however, is still a matter of intense debate. One way to examine these potential relationships is to use a nonhuman primate model for which complexities present in humans can be minimized. Recent research into the varying personality types of marmoset monkeys suggests that there are predominantly three to five core primary domains that most marmosets and other primates can be categorized into, such as dominance, sociability, and neuroticism. The aim of the proposed study was to categorize a small colony of marmosets into respective personality domains, and to examine correlations between the monkeys’ personalities and their cognitive ability. This study was be conducted on 27 marmoset monkeys (14 male, 13 female) housed in the Lacreuse lab at the University of Massachusetts Amherst. A personality survey based on Koski (2015) containing 55 personality traits was utilized by 8 human judges, all of whom have been working with these monkeys daily for at least one year. Each judge rated each individual monkey on each individual trait on a 1 to 7-point scale; 1 indicating total absence of a trait and 7 indicating extreme presence of a trait. Once the survey data was compiled, a principal component analysis (PCA) was conducted to condense the myriad of ratings into smaller distinguishable personality domains. Three personality types were identified in this population, consistent with other non-human primate species. An ICC(2) was performed to ensure the interrater reliabilities of the 8 judges were consistent enough to be considered. Lastly, a linear regression was conducted to reveal possible correlations between the observed personality domains and cognitive performance achieved in a reversal learning task. The results of this experiment showed no statistically significant relationships between any of the three personality domains: Assertiveness, Neuroticism, and Inquisitiveness with the reversal learning cognitive scores. Although these findings suggest that personality and cognitive flexibility are independent in marmosets, we cannot rule out that personality may influence other cognitive domains. Additional studies are needed to examine this possibility.

Orientador: Ricardo de Oliveira Orsi / Banca: Luis Carlos Marchini / Banca: Nabor Veiga / Resumo: Os objetivos do presente trabalho foram avaliar o efeito da própolis sobre o peso e parâmetros comportamentais de saguis Callithrix sp mantidos em cativeiro e submetidos ao estresse. Foram utilizados 26 animais da espécie Callithrix jacchus (sagüi-de-tufo-branco) e Callithrix penicillata (sagüi-de-tufo-preto), com idade entre 8 e 18 meses, de ambos os sexos, os quais foram divididos em três tratamentos: T1- dieta de rotina; T2- dieta de rotina acrescida de 2% de própolis bruta e T3- dieta de rotina acrescida de 5% de própolis bruta da alimentação em relação ao consumo diário estimado por indivíduo. Os animais foram alimentados durante sete dias com ou sem o acréscimo de própolis (momento 1). Após este período, os animais continuaram recebendo o mesmo esquema alimentar e foram submetidos ao estresse intruso x residente, durante sete dias (momento 2), totalizando 14 dias. Em ambos os momentos foram avaliados os parâmetros comportamentais (brincadeiras social e lúdica, vocalização, catação e deslocamento) e a manutenção do peso dos animais. De acordo com os resultados obtidos pode-se concluir que a adição da própolis na dieta de sagüis mantidos em cativeiro promoveu maior ganho de peso nos animais, embora não tenha apresentado efeito na redução do estresse, avaliado através de parâmetros comportamentais. Sugere-se, no entanto, a necessidade de novas pesquisas no sentido de estudar outras concentrações de própolis e maiores tempos de observação dos animais. / Abstract : The aim of the present work was to evaluate the effects of propolis on the weight and behavioral parameters of Callithrix sp marmosets kept in captivity and subjected to stress. Twenty-six Callithrix jacchus (white-tuffed-ear marmoset) and Callithrix penicillata (black-tuffed ear marmoset), aged between 8 and 18 months, males and females, were divided into three treatment groups: T1- routine diet; T2- routine diet added of 2% crude propolis and T3- routine diet added of 5% crude propolis from daily diet, in relation to estimated daily consumption per individual. Animals were fed during seven days with or without the addition of propolis (moment 1). After this period, they continued receiving the same diet scheme and were subjected to intrusive x resident stress for seven days (moment 2), in 14 days of experiment. In both moments, behavioral parameters (social and playful activities, vocalization, catching and moving) and weight were evaluated. According to the obtained results, the addition of propolis to the diet of marmosets kept in captivity leads to greater weight gain in the animals although it had no effect on stress reduction, which was evaluated through behavioral parameters. Thus, there is the need for additional studies using other propolis concentrations and longer observation times. / Mestre

The use of Positive Reinforcement Training (PRT) for co-operation during routine husbandry and laboratory procedures is widely advocated as a means of promoting the welfare of nonhuman primates. However, while research originating in US zoos provide qualitative descriptions of how PRT may be used in the training of a wide variety of species, quantitative data and evidence to support the view that PRT reduces stress predominately comes from laboratory studies of primates whose training may have used other methods. Despite official guidelines, training is rarely carried out in the UK and the educational and wider organisational structures concerning training, present in the US are largely absent. The techniques used in the UK were assessed through detailed observations recorded when four stump-tailed macaques were trained to co-operate during venipuncture. Data recorded during training sessions showed that although food rewards were given, their delivery was slow and inconsistent. A certain amount of coercion was used which violates a principle of PRT which states that co-operation should be voluntary. The macaques showed increasing resistance to the process and a mild but detrimental effect on the subsequent behaviour of the study animals. When training resumed 18 months later there were considerable improvements in the techniques used. The macaques showed a greater willingness to participate and there were no significant changes in their behaviour when training days were compared to those when training did not take place. The behaviour of the macaques during venipuncture was judged to be arising from engineered compliance rather than voluntary co-operation. However, it was concluded that the technique observed, if carried out correctly, was a reasonable compromise between forced restraint and voluntary co-operation given the paucity of evidence showing the effectiveness of PRT for invasive procedures. However, it was also concluded that the use of coercion should be recognised and provide a focus for future refinement. The effectiveness and welfare implications ofPRT was assessed through the training of common marmosets to target and allow in-homecage weighing and to provide urine samples. It was found that the trained animals perfonned reliably and that time invested in training could be recouped through faster data collection. Following a period of training or increased positive contact with humans, observations of marmoset behaviour showed a decrease in stress related behaviours and an increase in allogrooming supporting the view that improved relations with humans had a beneficial effect. Following exposure to a mild stressor, trained marmosets showed no elevation in levels of urinary cortisol or stress related behaviours. Untrained animals showed increased levels of locomoting and selfscratching following exposure to the same stressor. It was concluded that PRT successfully reduced the stress associated with the presence of, and manipulation by, humans. Final recommendations were that training can promote the welfare of nonhuman primates and should be used in UK laboratories to a greater extent than is currently the case. However, the lack of educational opportunities for animal trainers in the UK needs to be addressed. It was also recommended that in light of the growing evidence showing the benefits that can arise from training and good relations with humans, the zero-handling policy practiced in many UK zoos should be reassessed.

Thioredoxin is a small ubiquitous oxido-reductase found in all species. The highly conserved active site, which facilitates thioredoxins redox activity, contains two redox active cysteine residues. Thioredoxin has numerous protein substrates to which it donates H+ ions and it can also function as a free radical scavenger. Through these activities thioredoxin is able to influence the redox state of not only its protein targets, but also the entire cellular environment. Thioredoxin has been implicated in many biological functions, and one mechanism by which it influences these functions is through interactions with a number of transcription factors including NF-kappa-B and p53. Thioredoxin also has numerous extracellular biological roles. It has been shown that thioredoxin is actively secreted from a number of normal and transformed cell lines including fibroblasts and activated B and T cells. This study investigates the role of thioredoxin in embryonic implantation and cancer cell metastasis, two physiological functions which rely on the same basic processes. Thioredoxin expression has previously been shown to be increased in many cancers. However it has not yet been established whether this increase is a causative or a side effect of the cancerous phenotype. Similarly thioredoxin expression has previously been shown to be increased during different phases of the oestrus cycle and pregnancy. This thesis describes the role of thioredoxin in embryonic implantation using a marmoset model. A thioredoxin cDNA was isolated and subsequently sequenced. Preliminary antibody experiments indicated that the anti human thioredoxin monoclonal antibodies available in our laboratory would recognise marmoset thioredoxin. Subsequently immunocytochemistry using anti human thioredoxin antibodies was carried out on sectioned marmoset uterus and embryonic tissue. The results indicated that thioredoxin is expressed by cells at the embryonic-maternal interface of early implantation sites. Further studies demonstrated that thioredoxin is also expressed and secreted by cultured blastocysts in vitro. This thesis also describes the role of thioredoxin in cancer cell metastasis. Results of this study indicate that thioredoxin is actively involved in facilitating the invasive phenotype of breast cancer cells. The two cell lines utilised were MCF-7, a well differentiated, relatively non-invasive breast cancer cell line; and MDA-MB-231, a poorly differentiated, highly invasive breast cancer cell line. The cell lines were transfected with thioredoxin sense, antisense and 1SS (encodes thioredoxin with both active cysteine residues mutated to serine residues and is thus redox inactive) constructs. The results demonstrate that when endogenous thioredoxin levels are increased, i.e. transfected with a sense thioredoxin construct, the invasive breast cancer cell line MDA-MB-231 becomes more invasive, conversely when endogenous levels are decreased, i.e. transfected with antisense or 1SS constructs, the invasive capacity of these cells decreases. However, when the endogenous level of thioredoxin was manipulated in the relatively non-invasive cell line MCF-7 very little effect was observed. Results also indicate that thioredoxin has the ability to act as a chemoattractant for actively invading breast cancer cells. Both of these functions appear to be dependent on thioredoxin's redox activity. Additional studies described in this thesis have shown that thioredoxin is involved in the regulation of Sp1 in vitro. Sp1 is a transcription factor known to regulate the transcription of a number of genes whose products are intimately involved in the invasive phenotype. The results in this study suggest that Sp1 DNA binding is regulated by thioredoxin such that when reduced by the enzyme its binding to DNA is facilitated. Results also indicate that Sp1 may regulate the transcription of thioredoxin by binding to Sp1 sites within the thioredoxin promoter.

Weißbüschelaffen (WBA) sind wissenschaftlich häufig genutzte Modelltiere für diverse Humanerkrankungen. Zur Gesunderhaltung dieser Primaten sind grundlegende diagnostische Blutparameter unverzichtbar. Bisher erhobene Daten zeichneten sich jedoch durch große Divergenz aus. Ob Veränderungen in Haltungsbedingungen einen Einfluss auf diese Blutparameter nehmen, ist bis heute unklar. Somit war ein Ziel dieser Arbeit die Erhebung aktueller hämatologischer und klinisch-chemischer Blutparameter von WBA. Zudem wurde der Einfluss der routinemäßigen Umsetzung in eine neue Behausung auf die erhobenen Parameter sowie den Kortisolspiegel im Kot untersucht. Des Weiteren leiden WBA in menschlicher Obhut rezidivierend an gastrointestinalen Erkrankungen, die mittels klinischer Standardparameter allein nicht diagnostizierbar sind. Dabei spielt vor allem die Sensitivität gegenüber Futtermittelinhaltsstoffen (z. B. Gluten) eine Rolle, welche ursächlich im Zusammenhang mit dem Wasting Marmoset Syndrome (WMS) diskutiert wird. Im zweiten Teil der vorliegenden Arbeit sollten deshalb die gastrointestinalen Erkrankungen von in menschlicher Obhut lebenden WBA ätiologisch beleuchtet werden, vor allem hinsichtlich einer möglichen Sensitivität gegenüber Gluten. Im ersten Teil dieser Studie wurden von 54 WBA hämatologische und klinischchemische Richtwerte erhoben. Die ermittelten hämatologischen Blutrichtwerte ähneln denen aus den achtziger Jahren, die Daten der klinischen Chemie nur bedingt: Die Richtwertbereiche von Laktatdehydrogenase, Alaninaminotransferase, Lipase sowie Alkalische Phosphatase und Gesamtbilirubin weichen von den ehemals erhobenen Daten ab. Zudem wurden in der vorliegenden Arbeit geschlechtsabhängige Unterschiede ermittelt: Weibliche Tiere wiesen signifikant höheres mittleres Erythrozytenvolumen und mittleren Hämoglobingehalt des Einzelerythrozyten auf als männliche Tiere, wohingegen bei diesen ein signifikant höheres Gesamt- und Low density lipoprotein- Cholesterol im Vergleich zu weiblichen Affen messbar war. Des Weiteren wurden 16 Tiere über einen vierwöchigen Zeitraum in eine neue Umgebung verbracht, bevor sie in ihre Heimatbehausung zurückkehrten. Durch diese Umsetzung war bei den untersuchten Tieren die Leuko- und Lymphozytenzahl auch vier Wochen nach der Umsetzung erniedrigt. Zeitgleich lag ein erhöhter Kortisolspiegel vor, der im Kot bestimmt wurde. Im zweiten Teil der Studie wurden anhand humandiagnostischer Standards IgAAntikörper (AK) gegen Gliadin (AGA), Gewebstransglutaminase (tTG), deamidiertes Gliadin (ADGA) sowie Glykoprotein 2 (AGP2A) im Plasma von 24 WBA mittels eines ELISAs während glutenhaltiger (Diät 1) und glutenfreier Ernährung (Diät 2) bestimmt. Dabei wurden die klinische Symptomatik von WMS und das Körpergewicht der Tiere ebenfalls untersucht. Zudem erfolgte die Analyse von Kotproben antikörperpositiver Tiere hinsichtlich Qualität und Trockenmassegehalt während Diät 2 und einer darauf folgenden glutenhaltigen Provokationsdiät. Die serologische Diagnostik ergab einen signifikanten Rückgang von AGA, AK gegen tTG und AGP2A während Diät 2 bei Tieren, die nach Diät 1 erhöhte Werte aufwiesen. Diät 2 führte zu einem Rückgang der klinischen Symptome und einer signifikanten Gewichtszunahme bei antikörperpositiven WBA. Die glutenhaltige Provokationsdiät ergab eine verminderte Kotqualität mit einem niedrigeren Trockenmassegehalt. Es wurden im Rahmen dieser Arbeit aktuelle, hämatologische und klinisch-chemische Blutrichtwerte des WBA erhoben. Der durch Umsetzung in eine neue Behausung bedingte Stress ist bei WBA bis vier Wochen lang nachweisbar. Es ist sinnvoll, dies in der zeitlichen Planung wissenschaftlicher Studien zu berücksichtigen, um das Wohlbefinden der Tiere vor Versuchsbeginn sicherzustellen und den Einfluss von Stress auf experimentelle Ergebnisse zu minimieren. Der Nachweis grundlegender, an der Pathogenese der Zöliakie beteiligter Antikörper, in Kombination mit den klinischen Symptomen, deutet auf Glutensensitivität mit ätiologischer Beteiligung an WMS bei WBA hin. Die glutenfreie Ernährung von WBA in menschlicher Obhut ist daher sinnvoll und empfehlenswert. / Common marmosets are often used as animal models for human diseases. For their health maintenance, diagnostic blood values are absolutely essential. Previously obtained reference values are characterized by great value-specific differences. Moreover, the influence of routine measures on these blood parameters, e. g. changes in housing conditions, has not been examined yet. Therefore, the first aim of the present study was to update haematological and clinical chemical blood parameters of common marmosets. Further, the influence of stress, caused by relocation to a new housing, on these parameters and the cortisol level in feces was examined. In addition to that, common marmosets under human management are often affected by gastrointestinal diseases, which are difficult to diagnose with basic standard blood values. In this context, sensitivity to nutritional elements, e. g. gluten, plays an important role and is discussed as a potential cause of wasting marmoset syndrome (WMS). In the second part of this study, the recurrent gastrointestinal diseases of common marmosets under human management were aetiologically investigated, with special regard to possible gluten sensitivity. In the first part of this study, blood samples were obtained from 54 female and male common marmosets to evaluate standard values of haematology and clinical chemistry. The determined haematological parameters are similar to the already obtained data, the clinical chemistry values differ somewhat: The enzyme activities of lactate dehydrogenase, alanine aminotransferase and lipase in addition to the ranges of alkaline phosphatase and total bilirubin diverge from the data ascertained in this study. Moreover, female animals presented significantly higher mean corpuscular volume and mean corpuscular haemoglobin than males, whereas male common marmosets showed significantly higher total- and low density lipoprotein-cholesterol, compared to females. Further, 16 animals were relocated to a new environment for a time period of four weeks, before they returned to their home cages. The change of housing caused a decreased leuko- and lymphocyte count in all examined animals that was still measurable four weeks after the relocation. At the same time, an increased fecal cortisol level was determined. The aim of the second study was to investigate the modification of plasma antibodies to gliadin (AGA), tissue transglutaminase (tTG), deamidated gliadin (ADGA) and glycoprotein 2 (AGP2A) during two successive diets in 24 animals: A gluten-containing diet (diet 1) and a gluten-free diet (diet 2). Further, clinical symptoms of WMS and the animals’ body weight were also examined. An analysis of the feces of antibody-positive animals regarding changes in quality and dry matter content was carried out with samples collected during diet 2 and a successive gluten challenge diet of two months duration. The serological diagnostics resulted in a significant decline of AGA, antibodies to tTG and AGP2A during diet 2 in animals that had shown increased antibody concentrations during diet 1. Diet 2 also caused an amelioration of clinical symptoms and an increased body weight in antibody-positive animals. The gluten challenge resulted in a decreased feces quality and a lower fecal dry matter, compared to fecal samples of diet 2. In the context of this dissertation, parameters of haematology and clinical chemistry of the common marmoset were updated. Stress caused by relocation to a new housing was still measurable for a period of four weeks. It is therefore essential to consider this time span in the design of scientific studies to secure animal welfare prior to the study and to reduce the influence of stress on experimental results. In combination with the clinical symptoms, the detection of antibodies that are part of the pathogenesis of coeliac disease in humans strongly suggests gluten sensitivity with an aetiological connection to WMS in common marmosets. Therefore, gluten-free nutrition of common marmosets under human management is highly recommendable.

abstract: Head movement is a natural orienting behavior for sensing environmental events around us. Head movement is particularly important for identifying through the sense of hearing the location of an out-of-sight, rear-approaching target to avoid danger or threat. This research aims to design a portable device for detecting the head movement patterns of common marmoset monkeys in laboratory environments. Marmoset is a new-world primate species and has become increasingly popular for neuroscience research. Understanding the unique patterns of their head movements will improve its values as a new primate model for uncovering the neurobiology of natural orienting behavior. Due to their relatively small head size (5 cm in diameter) and body weight (300-500 g), the device has to meet several unique design requirements with respect to accuracy and workability. A head-mount wireless tracking system was implemented based on inertial sensors that are capable of detecting motion in the Yaw, Pitch and Roll axes. The sensors were connected to the encoding station, which transmits wirelessly the 3-axis movement data to the decoding station at the sampling rate of ~175 Hz. The decoding station relays this information to the computer for real-time display and analysis. Different tracking systems, based on the accelerometer and Inertial Measurement Unit is implemented to track the head movement pattern of the marmoset head. Using these systems, translational and rotational information of head movement are collected, and the data analysis focuses on the rotational head movement in body-constrained marmosets. Three stimulus conditions were tested: 1) Alert, 2) Idle 3) Sound only. The head movement patterns were examined when the house light was turned on and off for each stimulus. Angular velocity, angular displacement and angular acceleration were analyzed in all three axes. Fast and large head turns were observed in the Yaw axis in response to the alert stimuli and not much in the idle and sound-only stimulus conditions. Contrasting changes in speed and range of head movement were found between light-on and light-off situations. The mean peak angular displacement was 95 degrees (light on) and 55 (light off) and the mean peak angular velocity was 650 degrees/ second (light on) and 400 degrees/second (light off), respectively, in response to the alert stimuli. These results suggest that the marmoset monkeys may engage in different modes of orienting behaviors with respect to the availability of visual cues and thus the necessity of head movement. This study provides a useful tool for future studies in understanding the interplay among visual, auditory and vestibular systems during nature behavior. / Dissertation/Thesis / Masters Thesis Bioengineering 2015

Made available in DSpace on 2014-12-17T15:36:51Z (GMT). No. of bitstreams: 1 DerlanAD.pdf: 578623 bytes, checksum: b1a56f1bf9e153912c46dc26566e416e (MD5) Previous issue date: 2007-09-21 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The competition for resources is one of the costs of group living. The scramble competition is considered an indirect type of competition, mainly associated with factors like group size and distribution of resources. Contest competition occurs when individuals compete directly for resources. In species that exibit this type of competition the establishment of dominance hierarchy can occur, resulting in differences on feeding and reproductive benefits for each member of the group. In these cases, aggressive and submissive behaviors are expected as a way to signal social status. The aim of this study was to investigate the effects of social hierarchy over food ingestion in Callithrix jacchus. Data recording was from September/2006 to March/2007, eight days by month, at Floresta Nacional de A?u do Instituto Chico Mendes de Biodiversidade. The observation time started at 05:00 AM and finished after the last animal was on the sleeptree. Analyses of aggressive interactions, behavioral profile and diet, reveals a lot of advantages for dominat animals in the study group. Dominant individuals had higher intake of animal matter that subordinates. The last ones, consumed fruits, exsudate and, eventually, explored itens that were not common to the diet. We suggest that dominance hiearchy enable the reproductive female to assure priority on access to food resources, a important caracteristc to supply tha costs to maintain tha high reproductive taxa of the specie. We also suggest that reproductive male, due to the participation on food transfer, had the forage efficience reduced / A vida em grupo implica em competi??o por recursos entre os indiv?duos. Competi??o do tipo Scramble ? considerada um modo indireto de competi??o, associado principalmente a fatores como tamanho do grupo e distribui??o dos recursos no ambiente. Competi??o do tipo Contest ocorre quando indiv?duos competem diretamente por recursos. Em esp?cies sujeitas a esse modo de competi??o pode ocorrer estabelecimento de hierarquia de domin?ncia, resultando em diferen?as nos benef?cios obtidos pelos indiv?duos no ?mbito alimentar e reprodutivo. Intera??es agressivas, bem como exibi??es de submiss?o, s?o esperadas como forma de sinalizar status social. Neste estudo investigamos o modo como a hierarquia social atua sobre o sucesso alimentar em Callithrix jacchus. De setembro de 2006 a mar?o de 2007 observamos um grupo selvagem de sag?is na Floresta Nacional de A?u do Instituto Chico Mendes de Biodiversidade, 8 dias por m?s. As observa??es tinham inicio ?s 05:00 e t?rmino ap?s todos os indiv?duos se recolherem ? ?rvore de dormir. A coleta de dados foi realizada atrav?s da t?cnica animal focal com registro instant?neo a cada minuto, durante 15 minutos consecutivos. Analisando as intera??es agon?sticas, perfil comportamental e perfil alimentar do grupo, verificamos que a hierarquia social em C. jacchus, al?m dos benef?cios reprodutivos, tamb?m confere uma s?rie de vantagens no ?mbito alimentar ao casal reprodutor. Em nosso estudo, intera??es agressivas, incluindo persegui??es, arranh?es e mordidas, foram pouco freq?entes de modo intragrupo. As rela??es de domin?ncia, portanto, foram mais evidentes atrav?s de comportamentos de submiss?o. N?o constatamos diferen?as significativas entre os animais em rela??o ? taxa de ingest?o total. Todavia, a an?lise do perfil alimentar (propor??o de mat?ria animal e vegetal na dieta) revelou que dominantes asseguraram maior ingest?o de mat?ria animal, enquanto subordinados utilizaram frutos, exsudado e, eventualmente, exploraram itens que n?o eram comuns ? dieta. Sugerimos que atrav?s da hierarquia social, a f?mea reprodutora assegura acesso preferencial a recursos alimentares, sendo esta uma importante caracter?stica para suprir os custos envolvidos com a manuten??o da alta taxa reprodutiva verificada para a esp?cie. O macho reprodutor, apesar de tamb?m assegurar acesso a recursos, tem sua efici?ncia de forrageio reduzida devido ? participa??o na partilha de alimentos direcionada aos filhotes

Endometriose (EM) ist eine häufige gynäkologische Erkrankung, die bei betroffenen Frauen unter anderem mit chronischen Unterleibsschmerzen und Unfruchtbarkeit einhergeht (VALLE 2002). Bisher war es bei den zur Forschung verwendeten Modelltieren für EM (z.B. Rhesusaffe, ZONDERVAN et al. 2004; Weißbüschelaffe, EINSPANIER et al. 2006) nicht möglich festzustellen, ob bei ihnen schmerzhafte Beeinträchtigungen durch die Erkrankung bestehen. Um die Auswirkungen neuer Therapeutika auf das Wohlbefinden der Patientinnen bewerten zu können, werden Methoden benötigt, mit denen EM bedingte Beeinträchtigungen der Modelltiere dargestellt werden können. Daher war es das Ziel dieser Studie, bei einem Primatenmodell für EM, dem Weißbüschelaffen, neue nicht invasive Testsysteme zu etablieren, die zur Darstellung von EM bedingten Schmerzen und Beeinträchtigungen geeignet sind. Unter der Annahme, dass schmerzhafte Erkrankungen das Verhalten (WALLACE et al. 1990), die Beweglichkeit (FLECKNELL 1986) sowie die kognitiven Fähigkeiten (SMITH et al. 2006) der betroffenen Tiere beeinträchtigen können, wurden drei nicht invasive Testsysteme auf ihre Eignung untersucht, Schmerzen bei an EM erkrankten Weißbüschelaffen im Vergleich zu Kontrolltieren darzustellen. Zur Untersuchung des Verhaltens wurde die Videoüberwachung, für die motorischen Fähigkeiten der Futterbaum (modifiziert nach ROBERTS et al. 1993) und für die kognitiven Fähigkeiten der Wisconsin General Test Apparatus (WGTA, HARLOW 1949) sowie der Futterbaum verwendet. Im ersten Abschnitt dieser Studie wurde das Normalverhalten von neun Weißbüschelaffenpaaren per Videokamera über den gesamten Tagesverlauf von zwölf Stunden aufgezeichnet und unter anderem in Bezug auf Aktivität, soziale und eigene Körperpflege sowie Futter- und Wasseraufnahme analysiert. Der Verlauf der Tagesaktivität zeigte drei Maxima zwischen 7:00 und 8:00 Uhr, 11:00 und 12:00 Uhr sowie 14:00 und 15:00 Uhr, dabei war die ansteigende Aktivität als Futtersuchverhalten vor den Mahlzeiten zu werten. Das im ersten Abschnitt der Studie dargestellte Aktivitätsmuster wurde im zweiten Abschnitt verwendet, um die Versuche mit WGTA und Futterbaum besser in den Tagesverlauf der Tiere einzuordnen und darüber ihre Kooperativität zu steigern. Die Tiere führten die Tests immer zur gleichen Tageszeit durch, deshalb wurde somit eine optimale Vergleichbarkeit und Homogenität der Ergebnisse gewährleistet. Bei der Auswertung der Videodokumentation im zweiten Abschnitt dieser Studie zeigte sich, dass erkrankte Weibchen ihren Partner im Gegensatz zu den Kontrolltieren gar nicht pflegen (p=0,029) und die Aktivität der erkrankten Weibchen zwar deutlich, aber nicht signifikant (p=0,057) verringert war. Diese verringerte Aktivität ist möglicherweise ein Hinweis auf Schmerzen der an EM erkrankten Weibchen, während die nicht vorhandene soziale Körperpflege den partnerschaftlichen Problemen betroffener Frauen entsprechen könnte. In den ersten beiden kognitiven Tests mit dem WGTA führten die erkrankten Weibchen signifikant weniger Versuche pro Tag durch als die Kontrolltiere (p=0,006/ p=0,008). Darüber hinaus benötigten die erkrankten Tiere signifikant mehr Versuche, um den ersten Test zu verstehen (p=0,008). Diese Unterschiede zu den Kontrolltieren ließen sich in den folgenden drei Versuchsabschnitten nicht mehr nachweisen. Daraus lässt sich ableiten, dass die Weibchen mit EM sich schlecht auf neue Anforderungen einstellen und sich weniger lange auf eine gestellte Aufgabe konzentrieren können. Nach der International Primatological Society (MC CANN et al. 2007) kann eine verminderte Fähigkeit, sich auf neue Situationen einzustellen, als Anzeichen für Beeinträchtigungen gewertet werden. Bei der Auswertung der Futterbaum Testreihen, in denen sowohl kognitive als auch motorische Fähigkeiten der Tiere mit einer Art „Kletterbaum“ überprüft wurden, ergaben sich demgegenüber keine signifikanten Unterschiede zwischen der EM-Gruppe und den Kontrolltieren. Zusammenfassend eignen sich die Videodokumentation und der WGTA zur Darstellung von Beeinträchtigungen bei an EM erkrankten Weißbüschelaffen. Die beiden Testsysteme können in folgenden pharmakologischen Studien verwendet werden, um erstmals die Auswirkungen neuer Therapeutika auf das Wohlbefinden der Modelltiere zu bewerten. Zusätzlich ermöglichen die Ergebnisse dieser Studie ein Refinement (RUSSELL und BURCH 1959), da die bisher verwendeten invasiven Methoden (Laparoskopie, Laparotomie) zur Bewertung des Verhaltens der EM Läsionen unter einer Therapie ergänzt und sogar ersetzt werden könnten.:Inhaltsverzeichnis Abkürzungsverzeichnis 1 Einleitung 1 2 Literaturübersicht 3 2.1 Endometriose 3 2.2 Tiermodelle für Endometriose 7 2.3 Methoden zur Darstellung von Beeinträchtigungen und Schmerzen bei Tieren 8 2.4 Der Weißbüschelaffe 10 3 Tiere, Material und Methoden 13 3.1 Tiere 13 3.2 Videodokumentation 16 3.3 Wisconsin General Test Apparatus (WGTA) 17 3.4 Futterbaum 20 3.5 Statistische Verfahren 22 4 Ergebnisse 24 4.1 Videodokumentation im ersten Studienabschnitt 24 4.2 Ergebnisse im zweiten Studienabschnitt 27 4.2.1 Videodokumentation 27 4.2.2 WGTA 29 4.2.3 Futterbaum 33 5 Diskussion 35 6 Zusammenfassung 41 7 Summary 43 8 Literaturverzeichnis 45 9 Anhang I 9.1 Paper „Behavioural tests as indicator for pain and distress in a primate endometriosis model“ eingereicht am 17.12.2009 bei Laboratory Animals I 9.2 Abstract zu dem Vortrag auf der 47. Wissenschaftlichen Tagung der Gesellschaft für Versuchstierkunde GV-SOLAS am 13.-15. September 2009 in Wien XIX 9.3 Abstract zu einem Poster, vorgestellt auf dem 7. Leipzig Research Festival for Life Sciences am 12. Dezember 2008 XX 9.4 Abstract zu dem Vortrag auf der 18. Tagung der DVG-Fachgruppe Physiologie und Biochemie am 9.-11. März 2008 in Leipzig XXI Danksagung / Endometriosis (EM) is a common gynecological disease, which is known to cause chronic pelvic pain and infertility in women (VALLE 2002). Up to now, it was not possible to assess, whether the animal models for research (e.g. rhesus macaque, ZONDERVAN et al. 2004; common marmoset, EINSPANIER et al. 2006) suffer from pain or impairments due to the disease. Therefore, new test systems are needed to obtain pain and discomfort in animal models for EM to enable the validation of new therapeutic agents with a view to the patients well being. It was the aim of this study, to establish new non invasive test systems to investigate signs of discomfort in an animal model for EM, the marmoset monkey. Assuming that painful diseases can influence the behaviour (WALLACE et al. 1990), the mobility (FLECKNELL 1986) and the cognitive abilities (SMITH et al. 2006) of animals, three non invasive test systems were reviewed for their ability to detect EM associated pain in common marmosets. They were based on behaviour (videotaping), mobility and exploratory behaviour (food tree, modified after ROBERTS et al. 1993) and cognitive abilities (Wisconsin General Test Apparatus (HARLOW 1949) and food tree). In the first part of this study, the daily activity patterns, allo- and autogrooming as well as water and food intake of nine common marmoset couples were monitored over a 12-hour light phase by video recording. The animals showed a trimodal course of activity per day with maxima from 7:00-8:00h, 11:00-12:00h and 15:00-16:00h. These activity maxima represented foraging behaviour, as they were followed by frequent food intake phases. The knowledge of the daily activity patterns allowed to optimize the experimental conditions for the tasks with the food tree and the Wisconsin General Test Apparatus (WGTA; HARLOW 1949) in the second part of this study. As every animal solved the tasks at the same time of day, the comparability and homogeneity of the results were optimized. By analysing the video documentation in the second part of this study, the females with EM, in contrast to the control females, did not show any social grooming behaviour (p=0.029). Furthermore, their activity level was almost significantly decreased (p=0.057). This reduced activity could indicate towards pain in the diseased females, while the lack of social grooming is similar to partnership problems in diseased women. The WGTA tasks revealed, that the females with EM performed significantly less trials per day in the first two settings (p=0.006/ p=0.008) and needed more trials to solve the first setting than the control animals (p=0.008). Those differences between diseased females and control animals were not detectable in the following three settings of the WGTA tasks. These results demonstrate, that EM affected marmosets have difficulties to concentrate on cognitive tasks and to cope with new situations. According to the International Primatological Society (MC CANN et al. 2007), these difficulties to cope with new situations can be interpreted as signs of distress. The food tree, a kind of jungle gym, was used to assess the animals` cognitive abilities as well as their mobility, but there were no significant differences between the EM diseased females and the control animals. In conclusion, the videotaping and the WGTA are suitable methods to demonstrate signs for impairments due to EM in marmoset monkeys. In following pharmacological studies, both test systems will allow to evaluate the benefit of new therapeutic agents on the animal model`s well being. In addition, the results of this study can help to refine procedures by replacing invasive methods like laparotomy according to the Refinement of RUSSELL and BURCH (1959).:Inhaltsverzeichnis Abkürzungsverzeichnis 1 Einleitung 1 2 Literaturübersicht 3 2.1 Endometriose 3 2.2 Tiermodelle für Endometriose 7 2.3 Methoden zur Darstellung von Beeinträchtigungen und Schmerzen bei Tieren 8 2.4 Der Weißbüschelaffe 10 3 Tiere, Material und Methoden 13 3.1 Tiere 13 3.2 Videodokumentation 16 3.3 Wisconsin General Test Apparatus (WGTA) 17 3.4 Futterbaum 20 3.5 Statistische Verfahren 22 4 Ergebnisse 24 4.1 Videodokumentation im ersten Studienabschnitt 24 4.2 Ergebnisse im zweiten Studienabschnitt 27 4.2.1 Videodokumentation 27 4.2.2 WGTA 29 4.2.3 Futterbaum 33 5 Diskussion 35 6 Zusammenfassung 41 7 Summary 43 8 Literaturverzeichnis 45 9 Anhang I 9.1 Paper „Behavioural tests as indicator for pain and distress in a primate endometriosis model“ eingereicht am 17.12.2009 bei Laboratory Animals I 9.2 Abstract zu dem Vortrag auf der 47. Wissenschaftlichen Tagung der Gesellschaft für Versuchstierkunde GV-SOLAS am 13.-15. September 2009 in Wien XIX 9.3 Abstract zu einem Poster, vorgestellt auf dem 7. Leipzig Research Festival for Life Sciences am 12. Dezember 2008 XX 9.4 Abstract zu dem Vortrag auf der 18. Tagung der DVG-Fachgruppe Physiologie und Biochemie am 9.-11. März 2008 in Leipzig XXI Danksagung

Made available in DSpace on 2014-12-17T15:36:38Z (GMT). No. of bitstreams: 1 DijenaideCC_TESE.pdf: 3067760 bytes, checksum: 8bdbafb50f4741aa87b6e7ea5023d6a3 (MD5) Previous issue date: 2011-11-11 / The use of animal models in biomedical research is ever increasing. Models that use primates might also have advantages in terms of low maintenance costs and availability of biological knowledge, thereby favoring their use in different experimental protocols. Many current stress studies use animal models at different developmental stages since biological response differs during ontogeny. The aims of this study were to perform a detailed characterization of the developmental stages of common marmosets (Callithrix jacchus), a very important animal model used in biomedical research. Ten subjects, 6 females and 4 males, were followed from birth to initial adult age (16 months). Behavioral and fecal collection for measurement of adrenal (cortisol) and sex (progesterone, estradiol and androgens) hormones took place twice a week during the first month of life and once a week for the remainder of the study. Behavior was observed for 30 minutes in the morning (0700-09:00h) and afternoon (12:00-14:00h). Behavioral profile showed changes during ontogeny, characterizing the 4 developmental stages and the respective phases proposed by Le?o et al (2009).. Differentiation of developmental stages was considered using the onset, end, change and stabilization of the behavioral profile parental care (weaning and carrying), ingestion (solid food), affiliation (social grooming) and autogrooming, agonism (scent marking and piloerection) and play behavior and endocrine profile. Infant weaning and carrying terminated within the infantile stage and the peak of solid food ingestion was recorded in the infantile III phase. Receiving grooming was recorded earlier than grooming performed by the infant and autogrooming. The first episode of scent marking was recorded in the 4th week and it was the least variable behavior, in terms of its onset, which, in almost all animals, was between the 5th and 7th week of life. Solitary play and play with the twin started around the 7th week and play with other members of the group started 8 weeks later. Sex hormone secretion started to differ from basal levels between the 21st and 23rd week of life, in males and females, suggesting that puberty occurs simultaneously in both sexes. Basal cortisol, even at an early age, was higher in females than in males. However, cortisol was not correlated with the juvenile stage, as expected, since this stage corresponds to the transition between infancy and adult age and most behaviors are intensified by this time. The behavioral and endocrine profile of subadult animals did not differ from that of the adults. These results provide more detailed parameters for the developmental process of C. jacchus and open new perspectives for the use of experimental approaches focused on the intermediate ontogenetic phases of this species / O uso de modelos animais em pesquisa biom?dica ? cada vez mais crescente e os modelos utilizando primatas devem apresentar vantagens em termos de custos de manuten??o e caracter?sticas relacionadas ao maior conhecimento da sua biologia para uso em diferentes protocolos experimentais. Diferentes modelos atuais de pesquisa em estresse utilizam animais em diferentes est?gios do desenvolvimento e in?meros estudos demonstram a diferen?a na resposta biol?gica diante de agentes estressores ao longo da ontog?nese. Com o objetivo de caracterizar de maneira mais detalhada as fases do desenvolvimento do sagui comum, Callithrix jacchus, importante modelo utilizado em pesquisa biom?dica, utilizou-se 10 animais, 6 f?meas e 4 machos, que foram acompanhados desde o nascimento at? o in?cio da idade adulta (16 meses). A coleta de dados comportamentais e de fezes para a mensura??o dos horm?nios esteroides de origem adrenal (cortisol) e gonadais (progesterona, estradiol e andr?genos) foram realizadas duas vezes por semana no primeiro m?s de vida dos filhotes e semanalmente no restante do estudo. As observa??es comportamentais tiveram dura??o de 30 minutos e foram realizadas nos turnos matutino (07:00-09:00h) e vespertino (12:00-14:00h).O perfil comportamental de C. jacchus apresentou modifica??es ao longo da ontog?nese caracterizando cada um dos 4 est?gios ontogen?ticos e suas respectivas fases propostas na classifica??o de Le?o (2009). A diferencia??o das etapas do desenvolvimento foi feita a partir do surgimento, t?rmino, varia??o e estabiliza??o no perfil comportamental - cuidado parental (amamenta??o, transporte) ingest?o de alimentos (alimenta??o s?lida), afilia??o (cata??o social recebida e feita e autocata??o) agonismo (marca??o de cheiro e piloere??o) e brincadeira - e no perfil end?crino. O comportamento de amamenta??o e transporte terminou na fase infantil II e a ingest?o alimentar apresentou seu pico na fase infantil III. A cata??o social recebida antecedeu a cata??o feita e a autocata??o, e o comportamento de marca??o de cheiro foi o que apresentou menor variabilidade em rela??o ao seu in?cio, que se concentrou, em quase todos os animais, entre a 5? e 7? semanas de vida. A brincadeira solit?ria e com o g?meo principiaram ao redor da 7? semana e a brincadeira com outros membros do grupo se iniciou 8 semanas depois. A secre??o dos horm?nios sexuais passou a se diferenciar dos valores basais entre a 21? e 23? semanas em machos e f?meas, sugerindo que a puberdade acontece simultaneamente para os dois sexos. O cortisol basal de f?meas mesmo em idade imatura foi mais elevado do que nos machos. Contudo, as varia??es do cortisol n?o se correlacionaram com o est?gio juvenil como esperado uma vez que esse est?gio corresponde ? transi??o entre a idade infantil e a idade adulta, e que se expressou com a intensifica??o da maioria dos comportamentos. Os padr?es comportamentais e end?crinos dos adultos n?o diferiram entre as idades subadulta e adulta. Estes resultados disponibilizam par?metros de desenvolvimento mais detalhados para C. jacchus e abrem perspectivas para a utiliza??o de abordagens experimentais focadas em determinadas etapas da ontog?nese dessa esp?cie

Made available in DSpace on 2014-12-17T15:36:38Z (GMT). No. of bitstreams: 1 CrhistianeAS_TESE.pdf: 3719505 bytes, checksum: 0a7335581ac5a34358c53f3a78dc9a50 (MD5) Previous issue date: 2012-07-05 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The circadian system consists of multiple oscillators organized hierarchically, with the suprachiasmatic nucleus (SCN) as the master oscillator to mammalians. There are lots of evidences that each SCN cell is an oscillator and that entrainment depends upon coupling degree between them. Knowledge of the mechanism of coupling between the SCN cells is essential for understanding entrainment and expression of circadian rhythms, and thus promote the development of new treatments for circadian rhythmicity disorders, which may cause various diseases. Some authors suggest that the dissociation model of circadian rhythm activity of rats under T22, period near the limit of synchronization, is a good model to induce internal desynchronization, and in this way, enhance knowledge about the coupling mechanism. So, in order to evaluate the pattern of the motor activity circadian rhythm of marmosets, Callithrix jacchus, in light-dark cycles at the lower limit of entrainment, two experiments were conducted: 1) 6 adult females were submitted to the LD symmetric cycles T21, T22 and T21.5 for 60, 35 and 48 days, respectively; 2) 4 male and 4 female adults were subjected to T21 for 24 days followed by 18 days of LL, and then back to T21 for 24 days followed by 14 days of LL. Vocalizations of all animals and motor activity of each one of them were continuously recorded throughout the experiments, but the vocalizations were recorded only in Experiment 1. Under the Ts shorter than 24 h, two simultaneous circadian components appeared in motor activity, one with the same period of LD cycle, named light-entrained component, and the other in free-running, named non-light-entrained component. Both components were displayed for all the animals in T21, five animals (83.3%) in T21.5 and two animals (33.3%) in T22. For vocalizations both components were observed under the three Ts. Due to the different characteristics of these components we suggest that dissociation is result of partial synchronization to the LD cycle, wherein at least one group oscillator is synchronized to the LD by relative coordination and masking processes, while at least another group of oscillators is in free-running, but also under the influence of masking by the LD. As the T21 h was the only cycle able to promote the emergence of both circadian components in circadian rhythms of all Callithrix jacchus, this was then considered the lower entrainment limit of LD cycle promoter of dissociation in circadian rhythmicity of this species, and then suggested as a non-human primate model for forced desynchronization / O sistema circadiano ? formado por m?ltiplos osciladores organizados hierarquicamente, sendo o n?cleo supraquiasm?tico (NSQ) o oscilador principal nos mam?feros. H? v?rias evid?ncias de que cada c?lula do NSQ seja um oscilador e que a sincroniza??o resulte do grau de acoplamento entre elas. O conhecimento do mecanismo de acoplamento entre as c?lulas do NSQ ? essencial para se entender a sincroniza??o e a express?o dos ritmos circadianos e, dessa forma, propiciar o desenvolvimento de novos tratamentos para os dist?rbios da ritmicidade circadiana que podem gerar v?rias doen?as. V?rios autores sugerem o modelo de dissocia??o do ritmo circadiano da atividade motora de ratos sob T22, per?odo pr?ximo ao limite de sincroniza??o, como um bom modelo animal para promover dessincroniza??o interna, propiciando um maior conhecimento sobre o mecanismo de acoplamento. Ent?o, com o objetivo de avaliar o padr?o do ritmo circadiano de atividade motora de saguis, Callithrix jacchus, em ciclos claro-escuro no limite inferior de sincroniza??o, foram realizados dois experimentos: 1) 6 f?meas adultas foram submetidas aos ciclo CE sim?tricos T21, T21.5 e T22 durante 60, 35 e 48 dias, respectivamente; 2) 4 machos e 4 f?meas adultos foram submetidos a T21 por 24 dias, seguido de CC por 18 dias, depois voltaram ao T21 por mais 24 dias seguido de CC durante 14 dias. As vocaliza??es de todos os animais e a atividade motora de cada um foram registradas continuamente ao longo dos experimentos, sendo que as vocaliza??es foram registradas apenas no Experimento 1. Sob os Ts menores que 24 h foram observados dois componentes circadianos simult?neos na atividade motora, sendo um com o mesmo per?odo do ciclo CE, nomeado como componente sincronizado pela luz, e outro em livre-curso, nomeado como componente n?o sincronizado pela luz. Os dois componentes foram exibidos por todos os animais sob T21, cinco animais (83,3%) sob T21.5 e dois animais (33,3%) sob T22. Para as vocaliza??es foram observados os dois componentes sob os tr?s Ts. Devido ?s diferentes caracter?sticas desses componentes, sugerimos que a dissocia??o ? resultado de sincroniza??o parcial ao ciclo CE, onde pelo menos um grupo de osciladores est? sincronizado ao ciclo CE por processos de coordena??o relativa e mascaramento, enquanto pelo menos outro grupo de osciladores est? em livre-curso, mas tamb?m sob influ?ncia de mascaramento do CE. Como o T21 foi o ?nico a promover o surgimento dos dois componentes circadianos nos ritmos circadianos de todos Callithrix jacchus, este foi ent?o considerado o per?odo de ciclo CE promotor de dissocia??o na ritmicidade circadiana dessa esp?cie, sendo ent?o sugerido como modelo de dessincroniza??o for?ada em primatas n?o humanos diurnos