Dissertations / Theses on the topic 'Macropodidae'

Thesis (Ph.D.) -- University of Western Sydney, 2002.
"A thesis submitted to the University of Western Sydney in fulfilment of the requirements for the degree of Doctor of Philosophy" Bibliography : leaves 400-437.

Leukocytes play a central role in protecting the body against infectious organisms and their research is essential for understanding the mechanisms of immunity. By studying leukocytes across a range of species, insights are provided into differing strategies employed to ensure resistance to disease. Surprisingly, the structure and function of marsupial leukocytes has received very limited study. Marsupials represent a major evolutionary pathway with distinct differences in reproduction and development from placental mammals. These differences in the life history of marsupials place unique challenges on the immune system, and differences in leukocyte structure and function could be reasonably expected. In this thesis, studies were undertaken to examine the cytochemical, ultrastructural and functional features of leukocytes from species of marsupials, belonging to the family Macropodidae (kangaroos and wallabies). The aim of these studies was to elucidate the characteristics of macropodid leukocytes and to compare and contrast these features with the known characteristics of other mammalian leukocytes. Leukocytes from two species of macropodid, the tammar wallaby (Macropus eugenii) and the western grey kangaroo (Macropus fuliginosis), formed the basis of this study with additional material provided from quokka (Setonix brachyurus), woylie (Bettongia pencillata) and red kangaroo (Macropus rufus). Staining characteristics of cells were examined following reaction with Sudan black B, peroxidase, chloroacetate esterase, naphthyl butyrate esterase, alkaline phosphatase and periodic acid-Schiff. Peroxidase and Sudan Black B reactions were similar to domestic animal species but chloroacetate esterase and naphthyl butyrate esterase were unreliable as markers for macropodid neutrophils and monocytes, respectively. Significant variation in staining for alkaline phosphatase was seen between species of macropodid. Tammar wallabies and quokka demonstrated strong neutrophil alkaline phosphatase activity whereas western grey kangaroos, red kangaroos and woylies contained no activity within their leukocytes. Peroxidase and alkaline phosphatase cytochemistry were also assessed at the ultrastructural level with transmission electron microscopy. This allowed the identification of distinct granule populations within macropodid neutrophils. Two subcellular compartments containing alkaline phosphatase activity were identified within tammar wallaby neutrophils. These were considered equivalent to secretory vesicles and a subpopulation of specific granules. Tubular vesicles containing alkaline phosphatase were also identified within the eosinophils of tammar wallabies. These structures were a novel finding having not been reported previously in the eosinophils of other animal species. In addition to cytochemistry, the general ultrastructure of leukocytes from tammar wallabies and western grey kangaroos were reported. Results were similar to previous reports for other marsupial species. The current body of knowledge was extended by the first detailed description of the ultrastructure of basophils in a marsupial. To assess functional aspects of macropdid neutrophils, flow cytometric assays were performed examining oxidative burst responses and phagocytosis. Reactive oxygen species were generated by neutrophils from tammar wallabies and western grey kangaroos in response to phorbol 12-myristate 13-acetate but not N-formyl-Met-Leu-Phe or opsonised bacteria. Phagocytosis of opsonised bacteria was also measured in neutrophils from tammar wallabies, which was poor in contrast to ovine neutrophils. However, flow cytometric studies were limited by sample preparation. Further optimisation of isolation methods for tammar wallaby leukocytes should be undertaken before dogmatic conclusions are drawn. Overall, the results of this thesis demonstrate that, in the areas examined, the general characteristics of leukocyte structure and function of mammals are present in macropodids. However differences were identified both within and outside of the macropodid group. These differences have important ramifications for the use of ‘model’ species in the study of leukocyte biology in marsupials. The results also provide potentially useful tools for the clinical diagnosis of haematological disease in macropodids and may be of interest to those studying comparative and evolutionary aspects of leukocyte structure and function.

Leukocytes play a central role in protecting the body against infectious organisms and their research is essential for understanding the mechanisms of immunity. By studying leukocytes across a range of species, insights are provided into differing strategies employed to ensure resistance to disease. Surprisingly, the structure and function of marsupial leukocytes has received very limited study. Marsupials represent a major evolutionary pathway with distinct differences in reproduction and development from placental mammals. These differences in the life history of marsupials place unique challenges on the immune system, and differences in leukocyte structure and function could be reasonably expected. In this thesis, studies were undertaken to examine the cytochemical, ultrastructural and functional features of leukocytes from species of marsupials, belonging to the family Macropodidae (kangaroos and wallabies). The aim of these studies was to elucidate the characteristics of macropodid leukocytes and to compare and contrast these features with the known characteristics of other mammalian leukocytes. Leukocytes from two species of macropodid, the tammar wallaby (Macropus eugenii) and the western grey kangaroo (Macropus fuliginosis), formed the basis of this study with additional material provided from quokka (Setonix brachyurus), woylie (Bettongia pencillata) and red kangaroo (Macropus rufus). Staining characteristics of cells were examined following reaction with Sudan black B, peroxidase, chloroacetate esterase, naphthyl butyrate esterase, alkaline phosphatase and periodic acid-Schiff. Peroxidase and Sudan Black B reactions were similar to domestic animal species but chloroacetate esterase and naphthyl butyrate esterase were unreliable as markers for macropodid neutrophils and monocytes, respectively. Significant variation in staining for alkaline phosphatase was seen between species of macropodid. Tammar wallabies and quokka demonstrated strong neutrophil alkaline phosphatase activity whereas western grey kangaroos, red kangaroos and woylies contained no activity within their leukocytes. Peroxidase and alkaline phosphatase cytochemistry were also assessed at the ultrastructural level with transmission electron microscopy. This allowed the identification of distinct granule populations within macropodid neutrophils. Two subcellular compartments containing alkaline phosphatase activity were identified within tammar wallaby neutrophils. These were considered equivalent to secretory vesicles and a subpopulation of specific granules. Tubular vesicles containing alkaline phosphatase were also identified within the eosinophils of tammar wallabies. These structures were a novel finding having not been reported previously in the eosinophils of other animal species. In addition to cytochemistry, the general ultrastructure of leukocytes from tammar wallabies and western grey kangaroos were reported. Results were similar to previous reports for other marsupial species. The current body of knowledge was extended by the first detailed description of the ultrastructure of basophils in a marsupial. To assess functional aspects of macropdid neutrophils, flow cytometric assays were performed examining oxidative burst responses and phagocytosis. Reactive oxygen species were generated by neutrophils from tammar wallabies and western grey kangaroos in response to phorbol 12-myristate 13-acetate but not N-formyl-Met-Leu-Phe or opsonised bacteria. Phagocytosis of opsonised bacteria was also measured in neutrophils from tammar wallabies, which was poor in contrast to ovine neutrophils. However, flow cytometric studies were limited by sample preparation. Further optimisation of isolation methods for tammar wallaby leukocytes should be undertaken before dogmatic conclusions are drawn. Overall, the results of this thesis demonstrate that, in the areas examined, the general characteristics of leukocyte structure and function of mammals are present in macropodids. However differences were identified both within and outside of the macropodid group. These differences have important ramifications for the use of ‘model’ species in the study of leukocyte biology in marsupials. The results also provide potentially useful tools for the clinical diagnosis of haematological disease in macropodids and may be of interest to those studying comparative and evolutionary aspects of leukocyte structure and function.

Leukocytes play a central role in protecting the body against infectious organisms and their research is essential for understanding the mechanisms of immunity. By studying leukocytes across a range of species, insights are provided into differing strategies employed to ensure resistance to disease. Surprisingly, the structure and function of marsupial leukocytes has received very limited study. Marsupials represent a major evolutionary pathway with distinct differences in reproduction and development from placental mammals. These differences in the life history of marsupials place unique challenges on the immune system, and differences in leukocyte structure and function could be reasonably expected. In this thesis, studies were undertaken to examine the cytochemical, ultrastructural and functional features of leukocytes from species of marsupials, belonging to the family Macropodidae (kangaroos and wallabies). The aim of these studies was to elucidate the characteristics of macropodid leukocytes and to compare and contrast these features with the known characteristics of other mammalian leukocytes. Leukocytes from two species of macropodid, the tammar wallaby (Macropus eugenii) and the western grey kangaroo (Macropus fuliginosis), formed the basis of this study with additional material provided from quokka (Setonix brachyurus), woylie (Bettongia pencillata) and red kangaroo (Macropus rufus). Staining characteristics of cells were examined following reaction with Sudan black B, peroxidase, chloroacetate esterase, naphthyl butyrate esterase, alkaline phosphatase and periodic acid-Schiff. Peroxidase and Sudan Black B reactions were similar to domestic animal species but chloroacetate esterase and naphthyl butyrate esterase were unreliable as markers for macropodid neutrophils and monocytes, respectively. Significant variation in staining for alkaline phosphatase was seen between species of macropodid. Tammar wallabies and quokka demonstrated strong neutrophil alkaline phosphatase activity whereas western grey kangaroos, red kangaroos and woylies contained no activity within their leukocytes. Peroxidase and alkaline phosphatase cytochemistry were also assessed at the ultrastructural level with transmission electron microscopy. This allowed the identification of distinct granule populations within macropodid neutrophils. Two subcellular compartments containing alkaline phosphatase activity were identified within tammar wallaby neutrophils. These were considered equivalent to secretory vesicles and a subpopulation of specific granules. Tubular vesicles containing alkaline phosphatase were also identified within the eosinophils of tammar wallabies. These structures were a novel finding having not been reported previously in the eosinophils of other animal species. In addition to cytochemistry, the general ultrastructure of leukocytes from tammar wallabies and western grey kangaroos were reported. Results were similar to previous reports for other marsupial species. The current body of knowledge was extended by the first detailed description of the ultrastructure of basophils in a marsupial. To assess functional aspects of macropdid neutrophils, flow cytometric assays were performed examining oxidative burst responses and phagocytosis. Reactive oxygen species were generated by neutrophils from tammar wallabies and western grey kangaroos in response to phorbol 12-myristate 13-acetate but not N-formyl-Met-Leu-Phe or opsonised bacteria. Phagocytosis of opsonised bacteria was also measured in neutrophils from tammar wallabies, which was poor in contrast to ovine neutrophils. However, flow cytometric studies were limited by sample preparation. Further optimisation of isolation methods for tammar wallaby leukocytes should be undertaken before dogmatic conclusions are drawn. Overall, the results of this thesis demonstrate that, in the areas examined, the general characteristics of leukocyte structure and function of mammals are present in macropodids. However differences were identified both within and outside of the macropodid group. These differences have important ramifications for the use of ‘model’ species in the study of leukocyte biology in marsupials. The results also provide potentially useful tools for the clinical diagnosis of haematological disease in macropodids and may be of interest to those studying comparative and evolutionary aspects of leukocyte structure and function.

The quokka (Setonix brachyurus Quoy & Gaimard 1830) is a medium-sized, macropodid marsupial that is endemic to the mesic, south-western corner of Australia. While being a tourist icon on Rottnest Island, the species is threatened with extinction. It has been intensively studied on Rottnest Island in the 1960s and 1970s, however very little is known of its ecology on the mainland. Additionally the insular and mainland environments are extremely different suggesting that ecological differences between the two populations are likely. Consequently, this study sought to determine the basic autecology of the quokka and identify what factors have attributed to its threatened conservation status. The northern jarrah forest of Western Australia was selected as the study region due to it being at the northern limit of extant quokka distribution and because it was thought that the factors threatening the quokka would be exacerbated there. Fossil deposits suggest that the quokka originally occupied an area of approximately 49,000 km2 in the south-western corner of Australia. Historical literature show that they were widespread and abundant when Europeans colonised the region in 1829 but a noticeable and dramatic decline occurred a century later. The arrival of the red fox to the region coincided almost exactly with this decline and so it was probably ultimately responsible. Continued predation by both it and the feral cat are likely to have continued the decline, along with habitat destruction and modification through altered fire regimes. Specific surveys and literature searches show that since the 1950s, the area occupied by the quokka has declined by 45% and since 1990 by 29%. Based on the criteria of the IUCN (Hilton-Taylor 2000), the conservation status of the quokka should remain as vulnerable. An endangered status may be more applicable if the quokkas restriction to patches through its existence as a metapopulation is considered. Trapping of eight sites supporting quokka populations in the mid-1990s revealed three sites now locally extinct despite the ongoing, six year old, fox control programme. Another three are at serious risk of extinction. Extant population sizes ranged from one to 36 and population density ranged from 0.07 to 4.3 individuals per hectare. This is considered to be below the carrying capacity of each site. The overall quokka population size in the northern jarrah forest may be as low as 150 adult individuals, of which half are likely to be female. Even the largest extant populations are highly susceptible to stochastic extinction events. This small size was surprising considering the six year old, introduced predator control programme. Historically, the restriction to discrete habitat patches, the occasional inter-patch movement, the lack of correlation between the dynamics of each population and reports of frequent localised extinctions and colonisations suggest that the quokka population once existed as part of a classic metapopulation. The massive decline of the quokka in the 1930s pushed the metapopulation structure into a non-equilibrium state such that today, the extant populations are the terminal remnants of the original classic metapopulation. Wild mainland quokkas breed throughout the year. A significant reduction in the number of births occurs over summer and this coincides with a decline in female body weight. Despite this, the mainland quokka is relatively fecund and is able to wean two offspring per year. The level of recruitment from pouch young to independence was low and this may explain the apparent lack of population increase following the initiation of fox control. A total of 56 trapped quokkas were fitted with a radio collar. Mean home range size for quokkas was 6.39 ha with a core range of 1.21 ha and this was negatively related to population density. Male home ranges were larger than females but not significantly when the sexual size dimorphism was considered. Nocturnal ranges were larger than diurnal ranges reflecting nocturnal departures from the swamp refugia. Home range sizes varied seasonally, probably due to changes in the distance required to move to obtain sufficient nutrients and water over the dry summer compared to the wet winter and spring. Telemetry confirmed trapping results that showed no movement between swamps or populations. Home range centres shifted to the periphery of the swamp following the winter inundation and this may increase the species susceptibility to predation. The lack of dispersal is probably caused by quokka populations existing below carrying capacity and following selection for philopatry under the threat of predation for dispersing individuals. Without dispersal to recolonise or rescue unpopulated patches, the collapse of the original quokka metapopulation appears to have occurred. On a macrohabitat scale, the quokka in the northern jarrah forest is restricted to Agonis swamp shrubland habitats that form in the open, upper reaches of creek systems on the western side of the forest. This restriction was probably initially due to the high water requirements of the quokka but is likely to have been exacerbated by increased predation pressure since the arrival of the fox. On a microhabitat scale, the quokka is a habitat specialist, preferring early seral stage swamp habitats, probably for foraging, as part of a mosaic of old age swamp that provides refuge. Despite the six year old, introduced predator control programme, foxes and cats are still the major cause of mortality to quokkas. Road kills was the other identifiable cause. Individuals alive at the start of the study had an 81% chance of staying alive until the end. The likelihood of dying was minimised by grouping together with conspecifics, maximising home range size and maximising the time spent within the swampy refuge. Current rates of adult and juvenile survivorship should allow population recovery and so it seems pouch young mortality, reflected by low recruitment, has inhibited the anticipated population increase following predator control. The confounding effect of inadequate unbaited controls meant that little statistical evidence was available on the impact of introduced predators on the quokka, however the models provided support for earlier hypotheses of these. The presence of a quokka population at a site was related to the amount of poison baits delivered ??? reflecting predation pressure, the average age of the swamp and a mosaic of early and late seral stages within the swamp habitat. Recently burnt habitat is thought to provide food for quokkas and long unburnt habitat provides refuge from predation.

Despite extensive investigation, a complete understanding of the evolutionary history of the Macropodidae (kangaroos and their kin) has remained elusive. This research has utilized DNA sequences and retrotransposons (genes that jump around within the genome) to shed light on the evolutionary timing and dynamics of these iconic marsupials over the past 20 million years, and draw correlations with past climate change events. The research shows that these marsupials underwent a rapid radiation, diversifying into a wide array of forms, coincident with a trend of climatic cooling and drying over the past ~8 million years.

Lumholtz’s tree-kangaroo (Dendrolagus Iumholtzi), one of Australia’s largest folivores and one of only two tree-kangaroo species endemic to Australia and far north Queensland’s Wet Tropics. D. Iumholtzi are most commonly found in the fragmented rainforests that remain within an agricultural matrix in a relatively small area on the Atherton Tablelands. Unfortunately the majority of these fragments are on privately owned land and are not totally protected from clearing, therefore their long-term persistence is threatened by land clearing, further habitat fragmentation and mortality from dogs and cars. Although there have been a few studies on the ecology and habitat use of D. Iumholtzi, our current knowledge is limited. A more comprehensive understanding of spatial and floristic habitat use is essential for the conservation and management of D. Iumholtzi. This study examined the spatial organisation and habitat utilisation of Lumholtz’s tree-kangaroos in a Type lb rainforest fragment on the Atherton Tablelands and compared this to earlier studies (Procter-Gray 1985, Newell 1999). The two previous studies were both undertaken on the same spatially restricted rainforest type (Type Sb) only a couple of hundred metres apart, so this study has provided an important expansion of our understanding of D. Iumholtzi ecology across space and rainforest types. There were no significant effects of rainforest type on the home range sizes of D. Iumholtzi (Procter-Gray 1985, Newell 1999, This study). Male D. Iumholtziin this study held home ranges of 2.1 ± 0.7 ha (90% HM) overlapping that of several females but not other males, and females had exclusive home ranges of 2.1 ± 0.8 ha (90% HM) of a similar size to males. However, there was a large amount of variation in female home range sizes (0.1 —4.9 ha). Body weight did not explain this variation in home range sizes. This study also examined structural and floristic characteristics of the habitat and investigated if these could be used to model D. Iumholtzi habitat usage. The structural and floristic characters measured in this study could not be used to determine the focus of habitat usage. This study has shown that there is a more complex association between D. Iumholtzi and its use of habitat other than the structural characters of the habitat. D. Iumholtzi do select specific tree species, but there are strongly expressed individual preferences, similar to other arboreal folivores. The reasons for these specific choices are currently unclear but D. Iumholtzi are likely to choose trees for foliage characters, such as the levels of nutrients or plant defences, rather than for the species at a taxonomic level. This is also consistent with other arboreal folivores such as koalas and leaf-eating monkeys. The determination of which foliar characters are driving tree species or individual tree choice will require further research. This study tested and rejected a number of previous hypotheses regarding the characteristics determining D. Iumholtzi habitat use. They are not edge specialists, do not prefer regrowth or areas with a large variation in canopy height, or areas with high species diversity or density. The gastrointestinal morphology of D. Iumholtzi shares a number of features with other foregut fermenting folivores. Compared to other macropodids, D. Iumholtzi has a large sacciform forestomach and a large overall stomach capacity, and more similar in size and morphology to that of other arboreal foregut fermenting folivores, such as colobine monkeys. It is likely that these characteristics are adaptive for its diet of rainforest leaves. Lumholtz’s tree-kangaroos can be simply aged using a tooth wear index developed during this study. Aging is essential for establishing demographics, such as age specific mortality and fecundity of populations, currently unknown in D. Iumholtzi. Without the ability to age populations we cannot reliably undertake valuable estimations such as population viability analysis, which require these parameters. Additionally, this study has highlighted that not only one rainforest type is important to D. Iumholtzi and that more emphasis should be made on the preservation and restoration of all rainforest types. Furthermore, it is vital that all rainforest fragments including riparian zones, regrowth and corridors and stepping stones, should be conserved, rehabilitated and areas replanted as D. Iumholtzi habitat, as they are crucial to the species long term survival.

Lumholtz’s tree-kangaroo (Dendrolagus Iumholtzi), one of Australia’s largest folivores and one of only two tree-kangaroo species endemic to Australia and far north Queensland’s Wet Tropics. D. Iumholtzi are most commonly found in the fragmented rainforests that remain within an agricultural matrix in a relatively small area on the Atherton Tablelands. Unfortunately the majority of these fragments are on privately owned land and are not totally protected from clearing, therefore their long-term persistence is threatened by land clearing, further habitat fragmentation and mortality from dogs and cars. Although there have been a few studies on the ecology and habitat use of D. Iumholtzi, our current knowledge is limited. A more comprehensive understanding of spatial and floristic habitat use is essential for the conservation and management of D. Iumholtzi. This study examined the spatial organisation and habitat utilisation of Lumholtz’s tree-kangaroos in a Type lb rainforest fragment on the Atherton Tablelands and compared this to earlier studies (Procter-Gray 1985, Newell 1999). The two previous studies were both undertaken on the same spatially restricted rainforest type (Type Sb) only a couple of hundred metres apart, so this study has provided an important expansion of our understanding of D. Iumholtzi ecology across space and rainforest types. There were no significant effects of rainforest type on the home range sizes of D. Iumholtzi (Procter-Gray 1985, Newell 1999, This study). Male D. Iumholtziin this study held home ranges of 2.1 ± 0.7 ha (90% HM) overlapping that of several females but not other males, and females had exclusive home ranges of 2.1 ± 0.8 ha (90% HM) of a similar size to males. However, there was a large amount of variation in female home range sizes (0.1 —4.9 ha). Body weight did not explain this variation in home range sizes. This study also examined structural and floristic characteristics of the habitat and investigated if these could be used to model D. Iumholtzi habitat usage. The structural and floristic characters measured in this study could not be used to determine the focus of habitat usage. This study has shown that there is a more complex association between D. Iumholtzi and its use of habitat other than the structural characters of the habitat. D. Iumholtzi do select specific tree species, but there are strongly expressed individual preferences, similar to other arboreal folivores. The reasons for these specific choices are currently unclear but D. Iumholtzi are likely to choose trees for foliage characters, such as the levels of nutrients or plant defences, rather than for the species at a taxonomic level. This is also consistent with other arboreal folivores such as koalas and leaf-eating monkeys. The determination of which foliar characters are driving tree species or individual tree choice will require further research. This study tested and rejected a number of previous hypotheses regarding the characteristics determining D. Iumholtzi habitat use. They are not edge specialists, do not prefer regrowth or areas with a large variation in canopy height, or areas with high species diversity or density. The gastrointestinal morphology of D. Iumholtzi shares a number of features with other foregut fermenting folivores. Compared to other macropodids, D. Iumholtzi has a large sacciform forestomach and a large overall stomach capacity, and more similar in size and morphology to that of other arboreal foregut fermenting folivores, such as colobine monkeys. It is likely that these characteristics are adaptive for its diet of rainforest leaves. Lumholtz’s tree-kangaroos can be simply aged using a tooth wear index developed during this study. Aging is essential for establishing demographics, such as age specific mortality and fecundity of populations, currently unknown in D. Iumholtzi. Without the ability to age populations we cannot reliably undertake valuable estimations such as population viability analysis, which require these parameters. Additionally, this study has highlighted that not only one rainforest type is important to D. Iumholtzi and that more emphasis should be made on the preservation and restoration of all rainforest types. Furthermore, it is vital that all rainforest fragments including riparian zones, regrowth and corridors and stepping stones, should be conserved, rehabilitated and areas replanted as D. Iumholtzi habitat, as they are crucial to the species long term survival.

Morphological data are crucial in evolutionary analyses for merging fossils into the tree of life, calibrating dating analyses and for enhancing inference of biological patterns and processes. Morphological phylogenetics is dominated by homoplastic characters, functional and developmental correlations, and also by highly subjective definitions of characters and their states, which in turn can mislead phylogeny reconstruction. A first study assessed the implications of biases among characters in Mesozoic mammals. Then, geometric morphometrics and molecular data were combined to study the systematics of kangaroos and wallabies. Finally, new methodologies using 3D morphometrics and multivariate statistical analyses were developed for phylogenetic inference.

Marsupials and eutherians are the two principal groups of modern mammals. Mammalian immunological studies, to date, have focused on eutherian systems with little or no comprehensive work having been carried out on marsupials. This project investigates the functional and developmental aspects of T-cell responses in the marsupial, Macropus eugenii (Tammar wallaby) in both adults and pouch young at various stages of development. Determination of the age at which the Tammar wallaby immune system becomes competent has been examined through the use of cellular and molecular studies carried out on developing pouch young tissue. The capacity for generating an immunological response in adult and pouch young marsupials has been studied by following cellular proliferation in response to mitogens or mixed lymphocyte culture (MLC). After examining adult responses to mitogens and allogenic lymphocytes, optimised conditions were then used to examine the development of responsiveness in pouch young. Several further tests were conducted and findings shown. The study has shown that the earliest age at which Macropus eugenii is capable of mounting a T-cell mediated immune response is between 5 to 13 days post-partum

Dissertação de Mestrado Integrado em Medicina Veterinária
The genus Globocephaloides (Nematoda: Trichostrongyloidea) is a pathogenic group of parasitic nematodes present in the duodenum of kangaroos and wallabies (Marsupialia: Macropodidae) in Australia. Globocephaloides species (G. trifidospicularis, G. macropodis and G. affinis) have been poorly studied and thus, there are significant controversies regarding their systematics and population structures. In the present study, single-strand conformation polymorphism (SSCP) and targeted sequencing of the internal transcribed spacers (ITS) of nuclear ribosomal DNA, were used to assess the genetic variation within and among Globocephaloides populations and individuals, from different host species and geographical origins. No or minor (0.2%) variation was detected among individuals of G. trifidospicularis and G. affinis. However, within G. macropodis populations there was a consistent heterogeneity in the ITS sequences (5.2 - 7.1%) between worms derived from two different host species (Macropus agilis and M. dorsalis). Under light microscopy, these two G. macropodis genotypes differed by the length, arrangement and tip of the spicules, and by pattern of the bursal rays. Thus, they were considered to represent sibling species. The molecular and morphological evidence culminated with the erection of a new species, namely G. wallabiae, and provided further insights into the host affiliation and geographical ranges of Globocephaloides spp.: G. wallabiae occurs mainly in M. dorsalis (north/east), G. macropodis in M. agilis and Petrogale persephone (north), G. affinis in M. dorsalis (north-east), and G. trifidospicularis in various Macropus species (south). Moreover, in the present study, the phylogenetic analyses between the genus Globocephaloides and other trichostrongyloid genera, using divergent domains of the 28S rRNA genes, gave evidence for the exclusion of the genus Globocephaloides from the Herpetostrongylidae, with the suggestion of a new family, namely „Globocephaloididae‟, within the Heligmosomoidea.
RESUMO - CARACTERIZAÇÃO MOLECULAR E MORFOLÓGICA DO GÉNERO GLOBOCEPHALOIDES EM MARSUPIAIS MACROPODÍDEOS NA AUSTRÁLIA - O género Globocephaloides (Nematoda: Trichostrongyloidea) é um importante grupo de nemátodes patogénicos presente no duodeno de cangurus e wallabies (Marsupialia: Macropodidae) na Austrália. O estudo das espécies de Globocephaloides (G. trifidospicularis, G. macropodis e G. affinis) tem sido limitado, e por isso, existem numerosas controvérsias em relação à sua sistemática e às suas estruturas populacionais. No presente estudo, as técnicas de análise de ácidos nucleicos, „single-strand conformation polymorphism‟ (SSCP) e sequenciação-alvo da região ITS („internal transcribed spacers‟) do DNA ribossomal foram usadas com o objectivo de analisar a variação genética entre indivíduos e populações de Globocephaloides provenientes de diferentes hospedeiros e áreas geográficas. Em ambas as espécies G. trifidospicularis e G. affinis pouca ou nenhuma variação foi encontrada (0.2%). Contudo, nas populações de G. macropodis foi detectada uma consistente heterogeneidade nas sequências ITS (5.2 - 7.1%) entre espécimes provenientes de dois hospedeiros distintos (Macropus agilis e M. dorsalis). Recorrendo à microscopia óptica verificou-se que os dois genótipos de G. macropodis diferiam no comprimento, conformação e ponta das espículas, assim como, no padrão dos raios da bolsa copuladora. Como tal, foi considerado que os dois genótipos representavam duas espécies congéneres. Os resultados da biologia molecular e do estudo morfológico determinaram o reconhecimento de uma nova espécie denominada G. wallabiae e, além disso, forneceram dados sobre a especificidade de hospedeiros e distribuição geográfica do género Globocephaloides. G. wallabiae ocorre principalmente no hospedeiro M. dorsalis (norte/este), G. macropodis no M. agilis e Petrogale persephone (norte), G. affinis no M. dorsalis (nordeste) e G. trifidospicularis em várias espécies do género Macropus (sul). Adicionalmente, a análise das relações filogenéticas entre o género Globocephaloides e outros tricostrongilídeos, empregando os domínios do gene 28S rRNA, deu provas para a exclusão do género Globocephaloides da família Herpetostrongylidae, com a sugestão de uma nova família, denominada „Globocephalidae‟, inserida na superfamília Heligmosomoidea.
Financial support provided to the supervisors for the present study was from bodies including the Australian Research Council (ARC) [to Robin B. Gasser] and Australian Biological Resources Study (ABRS) [to Ian Beveridge]

In der vorliegenden Arbeit sollten die in der veterinärmedizinischen Literatur bisher diskutierten Ursachen für LJD bei Makropoden hinsichtlich ihrer tatsächlichen Bedeutung abgeklärt und die Eignung einer formalininaktivierten, bestandsspezifischen Adsorbatvakzine zur Prophylaxe von LJD getestet werden. Da LJD eine parodontale Erkrankung darstellt, wurden auch die für Entstehung einer humanen Parodontitis prädisponierenden Faktoren mit in die Untersuchung einbezogen. Es wurden Tupferproben zur bakteriologischen Untersuchung von insgesamt 15 gesunden und 11 an LJD erkrankten Kängurus entnommen. Dabei konnten gramnegative Anaerobier bei allen Tieren isoliert werden. Fusobacterium nucleatum wurde in 82% der von an LJD erkrankten und nur in 33% der von gesunden Tieren entnommenen Tupferproben nachgewiesen, womit sich ein signifikanter Zusammenhang (P < 0,05) zwischen diesem Erreger und LJD ergab. Weitere überwiegend bei erkrankten Makropoden nachgewiesene Anaerobier stellten Prevotella oris/oralis (bei 73% der LJD-Fälle und bei 40% der gesunden Tiere) sowie Capnocytophaga spp. (45% vs. 13%) dar. Bacteroides spp. und Porphyromonas gingivalis wurden – wenn auch nur mit 3 bzw. 2 Nachweisen – ausschließlich bei kranken Tieren isoliert. Fusobacterium necrophorum wurde jeweils in 27% der Kängurus gefunden und spielte damit in dieser Studie keine Rolle für die Entstehung von LJD. In Übereinstimmung mit der Literatur konnten Moraxella spp. ausschließlich bei gesunden Makropoden isoliert werden. Vertreter dieser Gattung gehören damit offensichtlich zur normalen Maulflora der Kängurus. Für die Zoos in Halle und Leipzig wurde eine formalininaktivierte, bestandsspezifische Adsorbatvakzine gegen die bei einem an LJD erkrankten Känguru des jeweiligen Bestandes isolierten gramnegativen Anaerobier hergestellt. 7 Tiere (2 Rote Riesenkängurus, 5 Bennettwallabies) des Leipziger Zoos und 6 Bennettkängurus des Zoos in Halle wurden geimpft, wobei Auffrischungsimpfungen nach 4 bzw. 8 Wochen und nach 6 bzw. 12 Monaten erfolgten. Die spezifischen AK gegen das Prüfantigen Fusobacterium necrophorum wurden im SLA bestimmt. Es konnte keine Erhöhung der AK-Titer induziert werden und auch die Todesrate infolge von LJD senkte sich während des Untersuchungszeitraumes von 42 Monaten in den beiden Zoos nicht. Die höchsten AK-Level (1:512 bis 1:2048) ließen sich im Serum von natürlich infizierten und letztendlich tödlich erkrankten Bennettwallabies des Zoos in Hoyerswerda feststellen. Der Nachweis von AK-Titern im Serum von nicht geimpften Jungtieren lässt vermuten, dass AK via Kolostrum oder Dottersackplazenta auf die Jungtiere übertragen werden. Die Untersuchungen hinsichtlich der Fütterung zeigten, dass im Zoo Leipzig eine azidotische Stoffwechsellage induziert wurde, was sich bei den Leipziger Bennettkängurus in einem mit 7,53 signifikant niedrigeren Vormagen-pH-Wert im Vergleich zu den Hallenser und Auer Tieren (8,25 und 8,38) offenbarte. Dies schlug sich auch in erhöhten K-, Cholesterol- und -Amylasewerten im Serum der Leipziger Wallabies nieder, womit gezeigt werden konnte, dass sich diese Parameter offenbar auch bei Makropoden zur Diagnostik einer chronischen Azidose eignen. Die Versorgung der Bennettkängurus in Magdeburg und Halle mit Ca und P war zwar nicht ausreichend, spiegelte sich aber nicht in veränderten Blutwerten dieser Mengenelemente wider. Die Aktivität der AP nimmt mit zunehmenden Alter ähnlich wie bei anderen Tierarten ab. Ihre negative Korrelation mit dem Alter der Tiere war dabei hochsignifikant (P < 0,001, r = 0,77 bzw. 0,62). Beim direkten Vergleich gesunder mit an LJD erkrankten Tieren konnte weder eine Störung im Ca/P-Stoffwechsel noch eine Azidose in Verbindung zu LJD gebracht werden. In allen Zoos erfolgte eine Überversorgung mit Vitamin A, wobei die Bedarfswerte für Schaflämmer um das 3,5fache bis 41fache übertroffen wurden. Den Bedarfswerten am nächsten lagen die Versorgungswerte der Bennettkängurus vom TP Aue und der Östlichen Grauen Riesenkängurus vom Zoo Magdeburg, beides Bestände ohne LJD. Die ermittelten Retinolplasmakonzentrationen standen in keiner Beziehung zu den Vitamin-A-Gehalten im Futter, was darauf hindeutet, dass sich Retinolbestimmungen im Blutplasma ebenso wie bei anderen Tierarten nur in extremsten defizitären Situationen zur Einschätzung des Vitamin-A-Status eignen. Ob eine Hypervitaminose A für die Entstehung von LJD tatsächlich eine Rolle spielt, muss in zukünftigen Arbeiten unter Einbeziehung von Retinolesterbestimmungen in der Leber abgeklärt werden. Die Glukosewerte lagen mit 8,57 mmol/l (M. rufus) bzw. 6,51 mmol/l (M. rufogriseus) über den bisher bekannten Werten aus der Literatur. Da die Werte bei an LJD erkrankten Kängurus niedriger waren als bei gesunden Tieren, kann ein Diabetes mellitus als Ursache für LJD ausgeschlossen werden. Weder die Durchsicht von 144 Sektionsprotokollen noch die Bestimmung der Kreatinin- und Harnstoffkonzentration im Serum von an LJD erkrankten Tieren ließen einen Zusammenhang zwischen Erkrankungen der Nieren und LJD erkennen. 30 Tiere verendeten an LJD, wovon 20% auch an den Nieren erkrankt waren. Allerdings wiesen auch 16,7% der anderweitig gestorbenen Kängurus eine Nierenerkrankung auf. Die Serumkonzentrationen von Harnstoff bzw. Kreatinin der an LJD erkrankten Makropoden unterschieden sich nicht von den für die gesunden Roten Riesenkängurus (7,40 mmol bzw. 114 mmol/l) und Bennettwallabies (7,81 mmol/l bzw. 86 mmol/l) ermittelten Werten. Insgesamt 184 Sera von 107 Kängurus wurden auf AK gegen MaHV-1 und MaHV-2 mittels Neutralisationtest geprüft. Während 94,4% bzw. 97,2% der Roten Riesenkängurus serologisch positiv für MaHV-1 bzw. MaHV-2 waren, reagierten von den 71 überprüften Bennettkängurus nur 4 bzw. 3 Tiere positiv. Unter den Wallabies befanden sich auch 21 an LJD erkrankte Tiere, wovon lediglich 2 Tiere gegen MaHV-1 und 1 Tier gegen MaHV-2 eine Serokonversion zeigten. Die AK-Titer der Roten Riesenkängurus ließen keine Unterschiede zwischen gesunden und an LJD leidenden Tieren zu und die entnommenen Serumpaarproben von 5 zum Zeitpunkt der Blutentnahme an LJD leidenden Riesenkängurus zeigten kein einheitliches Verhalten im Sinne einer Serokonversion. Somit ließ sich der Verdacht, dass die Reaktivierung latenter Herpesinfektionen die Ursache für LJD sein könnte, nicht bestätigen. Im Ergebnis der vorliegenden Studie und im Zusammenhang mit den Angaben aus der Literatur stellt sich LJD primär als eine Infektion mit gramnegativen Anaerobiern dar, wovon Fusobacterium nucleatum, Bacteroides spp., Prophyromonas gingivalis und Fusobacterium necrophorum, Biovar A die größte Bedeutung haben dürften. Den Abschluss der Arbeit bilden Empfehlungen für die Haltung von Kängurus in zoologischen Einrichtungen und für die Therapie von LJD. Im Anhang finden sich Röntgenaufnahmen und Photographien von erkrankten und gesunden Makropoden
The aim of this thesis was the investigation of the aetiology of Lumpy Jaw Disease (LJD) in macropods concentrating specifically on the causes of the diseases in current veterinary medicine literature and to evaluate the use of a group-specific Al(OH)3-adjuvanted, formalin-inactivated whole-cell vaccine for the control of LJD in kangaroos kept in zoos. LJD is regarded as periodontal disease, therefore the risk factors for the development of human periodontitis were also included in this study. The oral flora from 15 healthy macropods and 11 animals suffering from LJD was isolated. At least one anaerobic gram-negative bacterial species was found in swabs of each macropod. The occurrence of Fusobacterium nucleatum was associated with LJD (P < 0.05) by detecting this bacterium in 82% of the kangaroos suffering from LJD compared to only in 33% of the healthy animals. Prevotella oris/oralis and Capnocytophaga spp. were also predominantly found in diseased animals in comparison with healthy macropods (73% vs. 40% and 45% vs. 13% respectively). Bacteroides spp. and Porphyromonas gingivalis were isolated in only 3 and 2 kangaroos suffering from LJD, respectively. Contrary to previously published studies about LJD Fusobacterium necrophorum was not associated with LJD, as this anaerobe was detected in only 27% of the diseased as well as healthy macropods. Moraxella spp. seem to be a part of the normal oral flora of macropods and was found exclusively in healthy animals. 11 Red-necked Wallabies (Macropus rufogriseus) and 2 Red Kangaroos (Macropus rufus) were immunized with a group-specific Al(OH)3-adjuvanted, formalin-inactivated whole-cell vaccine containing previously in a kangaroo suffering from LJD isolated gramnegative anaerobs. The kangaroos were re-vaccinated after 1, 2, 6 and 12 months. Blood was collected from each animal at the same time. Antibodies were titrated against Fusobacterium necrophorum in an agglutination assay. The vaccine failed to induce increased levels of antibodies as well as to protect wallabies and kangaroos against LJD. As the highest antibody titres were detected in most severely diseased wallabies kept in the Hoyerswerda zoo, the protective role of the humoral immune response in LJD seems to be doubtful. The finding of detectable levels of antibodies in unvaccinated joeys supports the theory, that there is a transmission of antibodies from the mother to the offspring via colostrum or yolk-sac placenta. The diet of the Red-necked Wallabies in one zoo has induced an acidosis: The pH of the forestomach fluid collected by probang was lower in the animals of this zoo (pH = 7.53) than in the wallabies of two other zoos (pH = 8.25 and 8.38, respectively). Potassium, cholesterol and -amylase were also higher in the blood of the animals of this zoo in comparison to the wallabies of the two other ones, hence these blood values seem to be helpful for the diagnosis of chronic acidosis in macropods. There was a calcium and phosphor deficiency in the nutrition of the wallabies in two zoos, but the blood concentration of both of these minerals was not changed. The activity of the ALP correlated negative with the age of the Bennett`s Wallabies (P < 0.001, r = -.77 and r = -.62 respectively, depending on the instruments). All of the above mentioned blood values showed no differences between healthy and diseased animals and could so far not support the assumption, that an imbalance in Ca and P metabolism or an acidosis are important factors for LJD. The macropods of all investigated zoos were fed on a diet rich in vitamin A ranging from the 3.5 to the 41fold requirement for lambs. The vitamin A content of the diets for the 2 collections without a history of LJD was the lowest in this study. These results raised the point, that a hypervitaminosis A could be a more predisposing factor for LJD than a vitamin A deficiency. Due to the fact the plasma retinol concentration was independent from the vitamin A content of the diet and so not helpful in diagnosis of a vitamin A deficiency or toxicity, further investigations regarding the role of vitamin A in the aetiopathogenesis of LJD should include measurements of the liver tissue content of retinol esters. The glucose plasma concentration of the healthy Red Kangaroos (8.57 mmol/l) as well as the Red-necked Wallabies (6.51 mmol/l) was higher than previously published values for macropods, but also higher than the results of the diseased animals in this study. Therefore diabetes mellitus can be ruled out as an underlying factor for LJD. The analysis of 144 pathological records showed, that 30 animals died because of LJD, 20% of them and 16.7% of the other 114 macropods had a concurrent kidney disease. The urea and creatinin concentration in serum samples of healthy animals was not higher than the values of diseased animals. In conclusion, these results suggest kidney diseases are not important for the development of LJD. Altogether 184 sera collected from 107 kangaroos were tested for antibodies against MaHV-1 and MaHV-2 using a neutralisation assay. The prevalence of the MaHV-1- as well as MaHV-2-antibodies was high among the Red Kangaroos (94.4% and 97.2% respectively), but low among the Red-necked Wallabies (5.6% and 4.2% respectively). Seroconversion for MaHV-1 was seen in 2 out of 21 wallabies suffering from LJD, only 1 of these animals also had antibodies against MaHV-2. The antibody-titres against both of the macropodid herpes viruses also did not differ between Red Kangaroos with and without LJD, therefore a reactivation of a latent herpesvirus infection does not appear to be causative for LJD. In summary, considering the results of this study and previously published literature LJD is an infectious disease caused by gramnegative anaerobic bacteria with Fusobacterium nucleatum, Bacteroides spp., Porphyromonas gingivalis and Fusobacterium necrophorum subsp. necrophorum being of most significance. Recommendations concerning the keeping of kangaroos in captivity and the management of LJD are listed in the conclusion of this thesis. Some radiographs and photos of diseased and healthy kangaroos are attached

This thesis describes a comprehensive study of the cellular responses of a number of endangered marsupial species with a principal focus on the tammar wallaby (Macropus eugenii) as a model macropod species. The development of in vitro experimental assays for the assessment of immune responses in this model species are described, which provided a set of benchmarks for comparisons with other members of the Macropodidae and with eutherian mammals. Once this data was collected and protocols were established, the study was extended to include investigations of the immune responses in opportunistic samples obtained from the Rufous Hare-wallaby (Lagorchestes hirsutus), the Long-footed potoroo ( Potorous longipes) and the more common, but nonetheless still vulnerable, Long-nosed potoroo (Potorous tridactylus) with a view to investigating their apparent susceptibility to infection with intracellular pathogens, particularly mycobacterial species. The findings from the application of these assays suggest that the cellular immune responses of these species are relatively complex and involve a level of sophistication that rivals their eutherian counterparts. Specifically peripheral blood and tissue leukocytes were morphologically similar to those of other mammals, with the exception of tammar wallaby monocytes that appeared to contain few lysosomal granules, and the basophils of the Rufous Hare-wallaby that contained very large atypical granules. The overall findings of this study suggest that the immune systems of macropod species possess most of the sophistication associated with that of eutherian mammals. Whilst some differences were apparent in cells and their products in the test species, no single factor common to all macropods was identified as a cause for immune dysfunction. It appears likely that as yet undefined factors related to their confinement rather than an inherent defect in their immunocapacity is responsible for the apparent disease susceptibility of these animals.
Doctor of Philosophy (PhD)

This thesis describes a comprehensive study of the cellular responses of a number of endangered marsupial species with a principal focus on the tammar wallaby (Macropus eugenii) as a model macropod species. The development of in vitro experimental assays for the assessment of immune responses in this model species are described, which provided a set of benchmarks for comparisons with other members of the Macropodidae and with eutherian mammals. Once this data was collected and protocols were established, the study was extended to include investigations of the immune responses in opportunistic samples obtained from the Rufous Hare-wallaby (Lagorchestes hirsutus), the Long-footed potoroo ( Potorous longipes) and the more common, but nonetheless still vulnerable, Long-nosed potoroo (Potorous tridactylus) with a view to investigating their apparent susceptibility to infection with intracellular pathogens, particularly mycobacterial species. The findings from the application of these assays suggest that the cellular immune responses of these species are relatively complex and involve a level of sophistication that rivals their eutherian counterparts. Specifically peripheral blood and tissue leukocytes were morphologically similar to those of other mammals, with the exception of tammar wallaby monocytes that appeared to contain few lysosomal granules, and the basophils of the Rufous Hare-wallaby that contained very large atypical granules. The overall findings of this study suggest that the immune systems of macropod species possess most of the sophistication associated with that of eutherian mammals. Whilst some differences were apparent in cells and their products in the test species, no single factor common to all macropods was identified as a cause for immune dysfunction. It appears likely that as yet undefined factors related to their confinement rather than an inherent defect in their immunocapacity is responsible for the apparent disease susceptibility of these animals.

GnRH-targeted immunocontraception has potential as a non-lethal means of managing overabundant native species in Australia, and also represents a useful tool for studying the key processes within the reproductive endocrine axis. In this thesis, the effects of the GnRH vaccine 'GonaCon(TM)' on peri-pubertal development, growth, adult fertility and social behaviour are examined in two model macropodid species, the tammar wallaby, Macropus eugenii and the eastern grey kangaroo, M. giganteus. Vaccination against GnRH in adult male tammars resulted in rapid testicular atrophy and a loss of fertility for a minimum of two years in association with the production of significant GnRH antibody titres. Puberty was also delayed for a minimum of two years following vaccination of immature males of this species. Male-type growth rates were suppressed in association with depressed testosterone production in males of both age groups, and significant interruptions to both sexual and agonistic behaviours were observed in both adult- and juvenile-treated males. In females, GnRH vaccination effectively suppressed reproduction in 100% of adult female tammars over 5 breeding seasons. Treatment had no apparent effect on lactation but inhibited the production of young from diapausing blastocysts in this group, possibly by via some effect on the corpus luteum of pregnancy. Vaccinated sub-adult female eastern grey kangaroos failed to undergo normal peripubertal development (based on observations of pouch condition) and remained reproductively inactive for the duration of the study (2 1/2 years) - unlike Controls which all produced pouch young. No significant impacts on animal health or welfare were observed during this study however the effects of altered behavioural interactions will need to be re-evaluated on a larger scale to ascertain the potential for any negative impacts on animal welfare or the attainment of management objectives. Based on the long-lasting and highly efficacious results observed, GnRH-targeted immunocontraception would likely be a useful means of managing urban macropodid populations.

Interactions between multiple disturbances have been shown to have unexpected, and often undesirable effects on ecosystems and biodiversity. Improving our ability to predict and manage the outcomes of multiple disturbances is therefore an important research priority. In this thesis, I focused on the interaction between fire and grazing (or browsing) by large herbivores. Evidence of the individual effects of fire and large herbivores is substantial, but there has been little quantitative synthesis of the effects of native herbivores on biodiversity. Using a systematic review and meta-analysis, I found that high densities of large herbivores usually have negative effects on other animals. However, I found that interactions between large herbivores and episodic disturbances, such as fire, remain poorly understood. I therefore designed a field experiment to test the interactive effects of prescribed fire and large herbivores on forest flora and fauna. I used full and partial exclosure fences to create a gradient of herbivore pressure across both burnt and unburnt sites, and measured the responses of vegetation, spiders and small vertebrates. I found that fire and herbivory interacted strongly to affect vegetation, with herbivory limiting the recovery of vegetation from fire. In contrast to the vegetation response, small vertebrates responded to the individual, but not interactive effects of disturbance. I then focused on the mechanisms driving interactive effects on vegetation, and found that the interaction occurred through both numerically mediated (concentration of herbivores in burnt sites) and functionally moderated (stronger effect of herbivores post-fire) pathways. The differing responses of plants and animals to fire and herbivory was at odds with existing literature, where the effects of large herbivores on fauna are usually attributed to vegetation changes. I therefore tested to what extent vegetation mediated the effects of fire and herbivory on web-building spiders – a group sensitive to changes in habitat structure. Vegetation structure partially mediated the negative effect of fire on spider density, while negative effects of large herbivores on spiders were mostly independent of vegetation. Different x types of web builders differed in their responses, resulting in important changes to the spider community following disturbance. The results of my experiments highlight the importance of focusing on mechanistic pathways for understanding and managing disturbance interactions. However, in reviewing recently published fire-grazing studies, I found that most reported only net effects of interactions. I demonstrate how by failing to identify mechanistic pathways, or non-linear effects, such studies are limited in their management applications. I describe adjustments to disturbance interaction studies that would improve their ability to inform effective management and advance theory. Collectively, my research shows that fire and large herbivores can have strong interactive effects on forested ecosystems and their associated biota, and highlights the importance of considering large herbivores in forest fire planning. It also demonstrates the value of a mechanistic understanding of interactions for the management of disturbance regimes. Such considerations are of broad relevance to the management of multiple disturbances, particularly in the context of increasing global change.

In der vorliegenden Arbeit sollten die in der veterinärmedizinischen Literatur bisher diskutierten Ursachen für LJD bei Makropoden hinsichtlich ihrer tatsächlichen Bedeutung abgeklärt und die Eignung einer formalininaktivierten, bestandsspezifischen Adsorbatvakzine zur Prophylaxe von LJD getestet werden. Da LJD eine parodontale Erkrankung darstellt, wurden auch die für Entstehung einer humanen Parodontitis prädisponierenden Faktoren mit in die Untersuchung einbezogen. Es wurden Tupferproben zur bakteriologischen Untersuchung von insgesamt 15 gesunden und 11 an LJD erkrankten Kängurus entnommen. Dabei konnten gramnegative Anaerobier bei allen Tieren isoliert werden. Fusobacterium nucleatum wurde in 82% der von an LJD erkrankten und nur in 33% der von gesunden Tieren entnommenen Tupferproben nachgewiesen, womit sich ein signifikanter Zusammenhang (P < 0,05) zwischen diesem Erreger und LJD ergab. Weitere überwiegend bei erkrankten Makropoden nachgewiesene Anaerobier stellten Prevotella oris/oralis (bei 73% der LJD-Fälle und bei 40% der gesunden Tiere) sowie Capnocytophaga spp. (45% vs. 13%) dar. Bacteroides spp. und Porphyromonas gingivalis wurden – wenn auch nur mit 3 bzw. 2 Nachweisen – ausschließlich bei kranken Tieren isoliert. Fusobacterium necrophorum wurde jeweils in 27% der Kängurus gefunden und spielte damit in dieser Studie keine Rolle für die Entstehung von LJD. In Übereinstimmung mit der Literatur konnten Moraxella spp. ausschließlich bei gesunden Makropoden isoliert werden. Vertreter dieser Gattung gehören damit offensichtlich zur normalen Maulflora der Kängurus. Für die Zoos in Halle und Leipzig wurde eine formalininaktivierte, bestandsspezifische Adsorbatvakzine gegen die bei einem an LJD erkrankten Känguru des jeweiligen Bestandes isolierten gramnegativen Anaerobier hergestellt. 7 Tiere (2 Rote Riesenkängurus, 5 Bennettwallabies) des Leipziger Zoos und 6 Bennettkängurus des Zoos in Halle wurden geimpft, wobei Auffrischungsimpfungen nach 4 bzw. 8 Wochen und nach 6 bzw. 12 Monaten erfolgten. Die spezifischen AK gegen das Prüfantigen Fusobacterium necrophorum wurden im SLA bestimmt. Es konnte keine Erhöhung der AK-Titer induziert werden und auch die Todesrate infolge von LJD senkte sich während des Untersuchungszeitraumes von 42 Monaten in den beiden Zoos nicht. Die höchsten AK-Level (1:512 bis 1:2048) ließen sich im Serum von natürlich infizierten und letztendlich tödlich erkrankten Bennettwallabies des Zoos in Hoyerswerda feststellen. Der Nachweis von AK-Titern im Serum von nicht geimpften Jungtieren lässt vermuten, dass AK via Kolostrum oder Dottersackplazenta auf die Jungtiere übertragen werden. Die Untersuchungen hinsichtlich der Fütterung zeigten, dass im Zoo Leipzig eine azidotische Stoffwechsellage induziert wurde, was sich bei den Leipziger Bennettkängurus in einem mit 7,53 signifikant niedrigeren Vormagen-pH-Wert im Vergleich zu den Hallenser und Auer Tieren (8,25 und 8,38) offenbarte. Dies schlug sich auch in erhöhten K-, Cholesterol- und -Amylasewerten im Serum der Leipziger Wallabies nieder, womit gezeigt werden konnte, dass sich diese Parameter offenbar auch bei Makropoden zur Diagnostik einer chronischen Azidose eignen. Die Versorgung der Bennettkängurus in Magdeburg und Halle mit Ca und P war zwar nicht ausreichend, spiegelte sich aber nicht in veränderten Blutwerten dieser Mengenelemente wider. Die Aktivität der AP nimmt mit zunehmenden Alter ähnlich wie bei anderen Tierarten ab. Ihre negative Korrelation mit dem Alter der Tiere war dabei hochsignifikant (P < 0,001, r = 0,77 bzw. 0,62). Beim direkten Vergleich gesunder mit an LJD erkrankten Tieren konnte weder eine Störung im Ca/P-Stoffwechsel noch eine Azidose in Verbindung zu LJD gebracht werden. In allen Zoos erfolgte eine Überversorgung mit Vitamin A, wobei die Bedarfswerte für Schaflämmer um das 3,5fache bis 41fache übertroffen wurden. Den Bedarfswerten am nächsten lagen die Versorgungswerte der Bennettkängurus vom TP Aue und der Östlichen Grauen Riesenkängurus vom Zoo Magdeburg, beides Bestände ohne LJD. Die ermittelten Retinolplasmakonzentrationen standen in keiner Beziehung zu den Vitamin-A-Gehalten im Futter, was darauf hindeutet, dass sich Retinolbestimmungen im Blutplasma ebenso wie bei anderen Tierarten nur in extremsten defizitären Situationen zur Einschätzung des Vitamin-A-Status eignen. Ob eine Hypervitaminose A für die Entstehung von LJD tatsächlich eine Rolle spielt, muss in zukünftigen Arbeiten unter Einbeziehung von Retinolesterbestimmungen in der Leber abgeklärt werden. Die Glukosewerte lagen mit 8,57 mmol/l (M. rufus) bzw. 6,51 mmol/l (M. rufogriseus) über den bisher bekannten Werten aus der Literatur. Da die Werte bei an LJD erkrankten Kängurus niedriger waren als bei gesunden Tieren, kann ein Diabetes mellitus als Ursache für LJD ausgeschlossen werden. Weder die Durchsicht von 144 Sektionsprotokollen noch die Bestimmung der Kreatinin- und Harnstoffkonzentration im Serum von an LJD erkrankten Tieren ließen einen Zusammenhang zwischen Erkrankungen der Nieren und LJD erkennen. 30 Tiere verendeten an LJD, wovon 20% auch an den Nieren erkrankt waren. Allerdings wiesen auch 16,7% der anderweitig gestorbenen Kängurus eine Nierenerkrankung auf. Die Serumkonzentrationen von Harnstoff bzw. Kreatinin der an LJD erkrankten Makropoden unterschieden sich nicht von den für die gesunden Roten Riesenkängurus (7,40 mmol bzw. 114 mmol/l) und Bennettwallabies (7,81 mmol/l bzw. 86 mmol/l) ermittelten Werten. Insgesamt 184 Sera von 107 Kängurus wurden auf AK gegen MaHV-1 und MaHV-2 mittels Neutralisationtest geprüft. Während 94,4% bzw. 97,2% der Roten Riesenkängurus serologisch positiv für MaHV-1 bzw. MaHV-2 waren, reagierten von den 71 überprüften Bennettkängurus nur 4 bzw. 3 Tiere positiv. Unter den Wallabies befanden sich auch 21 an LJD erkrankte Tiere, wovon lediglich 2 Tiere gegen MaHV-1 und 1 Tier gegen MaHV-2 eine Serokonversion zeigten. Die AK-Titer der Roten Riesenkängurus ließen keine Unterschiede zwischen gesunden und an LJD leidenden Tieren zu und die entnommenen Serumpaarproben von 5 zum Zeitpunkt der Blutentnahme an LJD leidenden Riesenkängurus zeigten kein einheitliches Verhalten im Sinne einer Serokonversion. Somit ließ sich der Verdacht, dass die Reaktivierung latenter Herpesinfektionen die Ursache für LJD sein könnte, nicht bestätigen. Im Ergebnis der vorliegenden Studie und im Zusammenhang mit den Angaben aus der Literatur stellt sich LJD primär als eine Infektion mit gramnegativen Anaerobiern dar, wovon Fusobacterium nucleatum, Bacteroides spp., Prophyromonas gingivalis und Fusobacterium necrophorum, Biovar A die größte Bedeutung haben dürften. Den Abschluss der Arbeit bilden Empfehlungen für die Haltung von Kängurus in zoologischen Einrichtungen und für die Therapie von LJD. Im Anhang finden sich Röntgenaufnahmen und Photographien von erkrankten und gesunden Makropoden.
The aim of this thesis was the investigation of the aetiology of Lumpy Jaw Disease (LJD) in macropods concentrating specifically on the causes of the diseases in current veterinary medicine literature and to evaluate the use of a group-specific Al(OH)3-adjuvanted, formalin-inactivated whole-cell vaccine for the control of LJD in kangaroos kept in zoos. LJD is regarded as periodontal disease, therefore the risk factors for the development of human periodontitis were also included in this study. The oral flora from 15 healthy macropods and 11 animals suffering from LJD was isolated. At least one anaerobic gram-negative bacterial species was found in swabs of each macropod. The occurrence of Fusobacterium nucleatum was associated with LJD (P < 0.05) by detecting this bacterium in 82% of the kangaroos suffering from LJD compared to only in 33% of the healthy animals. Prevotella oris/oralis and Capnocytophaga spp. were also predominantly found in diseased animals in comparison with healthy macropods (73% vs. 40% and 45% vs. 13% respectively). Bacteroides spp. and Porphyromonas gingivalis were isolated in only 3 and 2 kangaroos suffering from LJD, respectively. Contrary to previously published studies about LJD Fusobacterium necrophorum was not associated with LJD, as this anaerobe was detected in only 27% of the diseased as well as healthy macropods. Moraxella spp. seem to be a part of the normal oral flora of macropods and was found exclusively in healthy animals. 11 Red-necked Wallabies (Macropus rufogriseus) and 2 Red Kangaroos (Macropus rufus) were immunized with a group-specific Al(OH)3-adjuvanted, formalin-inactivated whole-cell vaccine containing previously in a kangaroo suffering from LJD isolated gramnegative anaerobs. The kangaroos were re-vaccinated after 1, 2, 6 and 12 months. Blood was collected from each animal at the same time. Antibodies were titrated against Fusobacterium necrophorum in an agglutination assay. The vaccine failed to induce increased levels of antibodies as well as to protect wallabies and kangaroos against LJD. As the highest antibody titres were detected in most severely diseased wallabies kept in the Hoyerswerda zoo, the protective role of the humoral immune response in LJD seems to be doubtful. The finding of detectable levels of antibodies in unvaccinated joeys supports the theory, that there is a transmission of antibodies from the mother to the offspring via colostrum or yolk-sac placenta. The diet of the Red-necked Wallabies in one zoo has induced an acidosis: The pH of the forestomach fluid collected by probang was lower in the animals of this zoo (pH = 7.53) than in the wallabies of two other zoos (pH = 8.25 and 8.38, respectively). Potassium, cholesterol and -amylase were also higher in the blood of the animals of this zoo in comparison to the wallabies of the two other ones, hence these blood values seem to be helpful for the diagnosis of chronic acidosis in macropods. There was a calcium and phosphor deficiency in the nutrition of the wallabies in two zoos, but the blood concentration of both of these minerals was not changed. The activity of the ALP correlated negative with the age of the Bennett`s Wallabies (P < 0.001, r = -.77 and r = -.62 respectively, depending on the instruments). All of the above mentioned blood values showed no differences between healthy and diseased animals and could so far not support the assumption, that an imbalance in Ca and P metabolism or an acidosis are important factors for LJD. The macropods of all investigated zoos were fed on a diet rich in vitamin A ranging from the 3.5 to the 41fold requirement for lambs. The vitamin A content of the diets for the 2 collections without a history of LJD was the lowest in this study. These results raised the point, that a hypervitaminosis A could be a more predisposing factor for LJD than a vitamin A deficiency. Due to the fact the plasma retinol concentration was independent from the vitamin A content of the diet and so not helpful in diagnosis of a vitamin A deficiency or toxicity, further investigations regarding the role of vitamin A in the aetiopathogenesis of LJD should include measurements of the liver tissue content of retinol esters. The glucose plasma concentration of the healthy Red Kangaroos (8.57 mmol/l) as well as the Red-necked Wallabies (6.51 mmol/l) was higher than previously published values for macropods, but also higher than the results of the diseased animals in this study. Therefore diabetes mellitus can be ruled out as an underlying factor for LJD. The analysis of 144 pathological records showed, that 30 animals died because of LJD, 20% of them and 16.7% of the other 114 macropods had a concurrent kidney disease. The urea and creatinin concentration in serum samples of healthy animals was not higher than the values of diseased animals. In conclusion, these results suggest kidney diseases are not important for the development of LJD. Altogether 184 sera collected from 107 kangaroos were tested for antibodies against MaHV-1 and MaHV-2 using a neutralisation assay. The prevalence of the MaHV-1- as well as MaHV-2-antibodies was high among the Red Kangaroos (94.4% and 97.2% respectively), but low among the Red-necked Wallabies (5.6% and 4.2% respectively). Seroconversion for MaHV-1 was seen in 2 out of 21 wallabies suffering from LJD, only 1 of these animals also had antibodies against MaHV-2. The antibody-titres against both of the macropodid herpes viruses also did not differ between Red Kangaroos with and without LJD, therefore a reactivation of a latent herpesvirus infection does not appear to be causative for LJD. In summary, considering the results of this study and previously published literature LJD is an infectious disease caused by gramnegative anaerobic bacteria with Fusobacterium nucleatum, Bacteroides spp., Porphyromonas gingivalis and Fusobacterium necrophorum subsp. necrophorum being of most significance. Recommendations concerning the keeping of kangaroos in captivity and the management of LJD are listed in the conclusion of this thesis. Some radiographs and photos of diseased and healthy kangaroos are attached.