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No description available.

Lernaeocera branchialis (Linnaeus, 1767) is a parasitic copepod possessing a complex dual-host lifecycle. The “definitive” gadoid hosts, including Gadus morhua (Atlantic cod), Melanogrammus aeglefinus (haddock) and Merlangius merlangus (whiting), are infected by the fertilised female, which penetrates the host’s ventral aorta or bulbus arteriosus whilst undertaking extensive metamorphosis and a haematophagous lifestyle. The pathogenic effects of this activity upon the host have been well documented and mortality may occur, especially when multiple parasites are present. These negative impacts on cod, particularly juveniles, by L. branchialis have the potential to adversely affect cod aquaculture in the future, and already vulnerable wild cod stocks. This PhD project therefore, investigated the immune response of wild haddock and cultured-cod post-infection by L. branchialis, and the possible mechanisms by which the parasite modulates/evades the host’s immune response. The systemic immune response of both wild haddock and cultured-cod post-infection by L. branchialis depended on the maturation stage of the parasite, and in the former host species, upon the infection intensity. Wild haddock harbouring fully metamorphosed females showed an increase in circulating thrombocytes and a decrease in serum protein levels however; if multiple mature L. branchialis were present the haddock possessed reduced circulating monocytes, and increased circulating thrombocytes and serum anti-trypsin activity. Infection by L. branchialis was also associated with a suppressive effect on haddock serum spontaneous haemolytic activity. These responses were thought to be due to the host trying to counteract the increased damage caused by the massive increase in size and the feeding of the mature parasite, which is more pronounced when multiple parasites are present, resulting in the increase in some parameters and the ‘consumption’ of others. However, the effect of parasite-derived secretions and other pathogens due to observations on wild fish could not be discounted. The laboratory-infection of cultured-cod from two different sources was also performed in order to study the immune response over time. The two groups of cod showed differences in their immune response to L. branchialis. The first group showed suppressed respiratory burst activity of phagocytes, as the parasite reached the early penella sub-stage, whilst no suppression in phagocyte respiratory burst activity was found in the second group. The parasite was found to migrate along the afferent branchial artery of the cod where a thrombus formed and was present throughout its migration into the ventral aorta. At 14 d post-infection, leukocytes expressing Interleukin 8 mRNA were observed within the free-flowing blood at the periphery of the organising thrombus within the lumen of the ventral aorta. This was speculated to aid the recruitment and activation of leukocytes to the site, and the maturation and neovascularisation of granulation tissue. The infection of the second group subsided with the death of the parasite, and none of the parasites metamorphosed past the early penella sub-stage. The live parasites infecting the first group of cod did not possess IgM or complement component C3 binding on their cuticle, however, both IgM and C3 binding occurred on the dead parasites in the second infection trial. This may highlight the importance of these opsonins and the cytotoxic effect of phagocytes in the elimination of L. branchialis by some cod. However, the first infection was terminated as the parasite reached the early penella sub-stage due to a loss of stock cod prior to the study, so the long-term success of the infection can not be concluded. Therefore, the immune response to infection needs to be determined over the entire metamorphosis of L. branchialis to determine whether the infection was successful or not, and preferably in populations with varying susceptibility to L. branchialis. This will not be possible without further studies into the resistance of different stocks of cultured-cod. Many arthropod parasites, such as ticks and salmon lice, have been previously documented to produce pharmacologically active secretions, aiding host invasion and parasite feeding, preventing the host immune response from working effectively against the parasite, all aimed at improving survival of the parasite. Therefore, the effects of the secretory/excretory products (SEPs) produced during the initial infective stage and by the mature, fully metamorphosed female on the immune response of cultured-cod in vitro, and the location of exocrine glands associated with the oral region of the parasite were investigated. The SEPs from the infective stage of the parasite were found not to affect the intracellular hydrogen peroxide (H2O2) production of phagocytes. The practical difficulties in collecting large quantities of the SEPs from the infective stage meant that their effects could not be tested on the other host immune parameters studied. The SEPs from fully metamorphosed female L. branchialis, however, had a number of suppressive effects on the host immune response in vitro including: 1) suppression of the intracellular production of cytotoxic H2O2 during the respiratory burst of phagocytic leukocytes post-PMA stimulation, 2) suppression of the production of macrophage activating factor by leukocytes with a priming effect on naïve phagocyte function, and 3) suppression of the chemo-attraction ‘power’ of zymosan activated cod serum, i.e. anaphylatoxin activity, on head kidney-derived leukocytes. These effects were dose-dependent, and highlight the capacity of L. branchialis to suppress its host’s innate immune response at the local feeding area. Further work is required to establish the mechanisms by which the parasite-derived SEPs suppress these host immune parameters, and to identify which molecules produced by the parasite are responsible. The correlation between these in vitro results, and systemic immune parameters measured from laboratory-infected Atlantic cod and wild infected haddock are discussed. Host immuno-modulation by other arthropod parasites is mediated by pharmacologically active secretions produced by exocrine glands. Therefore, the exocrine glands of the infective and fully metamorphosed female L. branchialis were also investigated in order to identify those that might be responsible for the secretion of host-modifying products. Adult female exocrine glands were mapped using diaminobenzidine (DAB), most commonly known to stain peroxidases and catalases. These compounds are known to be involved in the neutralisation of harmful free radicals which are released during the respiratory burst and tissue damage. Such products may therefore be important protective secretory components at the site of feeding / infection. Exocrine glands were located in the infective stage associated with the oral region, one pair termed the anterior gland complex (AGC), and the other pair extending either side of the oral cone termed the circum-oral glands (CG). These were further investigated using light microscopy and transmission electron microcopy. The AGC and CGs possessed multi-component secretions and they possessed secretory vesicles, abundant and highly active rough endoplasmic reticulum and Golgi apparatus suggesting that protein is an important component of the secretory products. These glands were also observed in the fully metamorphosed females where they had increased in size within the cephalothorax post-metamorphosis. It is hoped that the identification of these glandular structures, which are thought to secrete within the local vicinity of the oral cone, will aid future studies regarding the identification and secretion kinetics of parasite-derived molecules during the infection and feeding process.

The intracellular parasite Trichinella is a genus in phylum Nematoda that contains six named species including Trichinella spiralis and Trichinella pseudospiralis. These parasites infect a large variety of wild and domestic animals, human beings and a few species of birds. The parasitic strategies and the pathological effects on the host between trichinella spiralis and Trichinella pseudospiralis are quite different. The purpose of this study is to gain an understanding of the physiological, immunological and pathological differences between these two species of Trichinella using infections in the mouse as a model. In the course of this research I have attempted to answer the following questions: A) Is cortisol a factor in the differences of the host response to Trichinella spiralis or Trichinella pseudospiralis? B) Are there differences in leukocyte response in the peripheral blood of Trichinella infected mice? C) Are there differences in the up-regulation or down-regulation of cell surface molecules on leukocytes in the spleens of Trichinella infected mice? D) Is there a difference in the degree of muscle damage (as measured by creatine phosphokinase) when infections by the two species of Trichinella are compared? E) Are there differences in angiogenesis and collagen deposition in Trichinella infected mice and are these differences related to cortisol? F) Is nitric oxide a component in parasite killing and are there differences in nitric oxide production in host mice when the two species of Trichinella are compared? My research has shown that there are significant differences in the parasitic strategies and pathological consequences in mice infected with one or the other of the two species of Trichinella. The two species appear to generate different immune and inflammatory responses from the host. Trichinella pseudospiralis is much less damaging to the host, generates a very different peripheral blood response, stimulates the production of substantially greater levels of serum cortisol, generates a significantly different profile in cytokine production presents a very different cell surface antigen profile and does not produce a collagen nurse cell or generate an angiogenic response when compared to T spiralis. In addition, I have shown a role for nitric oxide in parasite killing and a role for serum cortisol in larval survival. I have also shown that cortisol has no role in either collagen deposition or the angiogenic response in Balb/c mice under the experimental conditions detailed here.

Small broomrape, a holoparasitic weed, is a relatively new weed introduction in the Pacific Northwest that has contaminated a limited number of red clover fields in Oregon. Greenhouse and field studies were conducted to evaluate small broomrape response to common crop and weed species in the Pacific Northwest. Host species in the greenhouse or field study included alfalfa, arrowleaf clover, carrot, celery, common vetch, crimson clover, lettuce, prickly lettuce, red clover, spotted catsear, subterranean clover, white clover, and wild carrot. False-host species included barley, birdsfoot trefoil, creeping bentgrass, cucumber, field corn, fine fescue, flax, Italian ryegrass, nasturtium, oat, orchardgrass, perennial ryegrass, snap bean, sugar pea, sunflower, sweet corn, tall fescue, tomato, and wheat. Non-host species included sugar beet and curly dock. The greenhouse polyethylene bag system provided a rapid and inexpensive screening for plant species host status to small broomrape. Germination and attachment to host roots are initiated by chemical exudates, that may change concentration in response to nutrient availability and microorganisms. Red clover was grown in varying concentrations of ammonium sulfate fertilizer with and without Rhizobium inoculation, and with small broomrape seeds. Neither Rhizobium inoculation nor ammonium concentration influenced the number of small broomrape attachments to red clover roots. A survey was conducted of red clover seed growers with small broomrape-contaminated fields in the Pacific Northwest. Red clover seed from six respondents were cleaned at the same cleaning facility, and the same respondents purchased their seed stock from this cleaning facility. Small broomrape was not identified in red clover fields prior to or during the first clover seed harvest of fall planted red clover in small broomrape-contaminated sites. / Graduation date: 2003

Bibliography: leaves 196-210 / xi, 256, [85] leaves : (ill. (some col.) ; 31 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Entomology, Waite Agricultural Research Institute, 1985

Orientador: Wilson Castro Ferreira Junior / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Matemática, Estatística e Computação Científica / Made available in DSpace on 2018-08-20T05:05:37Z (GMT). No. of bitstreams: 1 Assis_RaulAbreude_D.pdf: 6617583 bytes, checksum: 80b80410952f2ac47edc7cf71466fb07 (MD5) Previous issue date: 2012 / Resumo: Neste trabalho apresentamos modelos matemáticos de processos evolutivos, utilizando como abordagem principal a descrição da frequência de fenótipos em populações. São propostos diversos modelos baseados em equações diferenciais parciais, ordinárias e equações de recorrência. São apresentados resultados de suas análises, bem como comparações com modelos genéticos aditivos. Como uma ilustração do tipo de abordagem proposto, criamos um modelo de um sistema do tipo parasita-hospedeiro (Maculinea-Myrmica) utilizando-o com o objetivo de analisar comportamento de especificidade de hospedeiro. Finalmente, apresentamos generalizações dos modelos em espaços de fenótipos e indicamos direções para aprofundamento de pesquisa / Abstract: In this thesis we present models of evolutionary dynamics that describe the changes in frequencies of phenotypes in populations. The models proposed are based on ordinary and partial differential equations and difference equations. Results from the analysis of the models and comparisons with the behavior of additive genetic models are presented. We develop a model for a host-parasite system (Maculinea-Myrmica) using the phenotypic space aprroach with the objective of analysing the hostspecificity behavior of the species. Finally, we present generalizations of the models of phenotypic spaces and indicate directions for further research in the area / Doutorado / Matematica Aplicada / Doutor em Matemática Aplicada

Human malaria is an economically important disease caused by single-celled parasites of the Plasmodium genus whose biology displays great evolutionary adaptation to both its mammalian host and transmitting vectors. This thesis details the 70 kDa heat shock protein (Hsp70) and J protein chaperone complements in malaria parasites affecting humans, primates and rodents. Heat shock proteins comprise a family of evolutionary conserved and structurally related proteins that play a crucial role in maintaining the structural integrity of proteins during normal and stress conditions. They are considered future therapeutic targets in various cellular systems including Plasmodium falciparum. J proteins (Hsp40) canonically partner with Hsp70s during protein synthesis and folding, trafficking or targeting of proteins for degradation. However, in P. falciparum, these classes of proteins have also been implicated in aiding the active transport of parasite proteins to the erythrocyte cytosol following erythrocyte entry by the parasite. This host-parasite “cross-talk” results in tremendous modifications of the infected erythrocyte, imparting properties that allow it to adhere to the endothelium, preventing splenic clearance. The genome of P. falciparum encodes six Hsp70 homologues and a large number of J proteins that localize to the various intracellular compartments or are exported to the infected erythrocyte cytosol. Understanding the Hsp70-J protein interactions and/or partnerships is an essential step for drug target validation and illumination of parasite biology. A review of these chaperone complements across the Plasmodium species shows that P. falciparum possesses an expanded Hsp70-J protein complement compared to the rodent and primate infecting species. It further highlights how unique the P. falciparum chaperone complement is compared to the other Plasmodium species included in the analysis. In silico analysis showed that the genome of P. falciparum encodes approximately 49 J proteins, 19 of which contain a PEXEL motif that has been implicated in routing proteins to the infected erythrocyte. Most of these PEXEL containing J proteins are unique with no homologues in the human system and are considered as attractive drug targets. Very few of the predicted J proteins in P. falciparum have been experimentally characterized. To this end, cell biological and biochemical approaches were employed to characterize PFB0595w and PFD0462w (Pfj1) J proteins. The uniqueness of Pfj1 and the controversy in literature regarding its localization formed the basis for the experimental work. This is the first study showing that Pfj1 localizes to the mitochondrion in the intraerythrocytic stage of development of P. falciparum and has further proposed PfHsp70-3 as a potential Hsp70 partner. Indeed, attempts to heterologously express and purify Pfj1 for its characterization are described. It is also the first study that details the successful expression and purification of PfHsp70-3. Further, research findings have described for the first time the expression and localization of PFB0595w in the intraerythrocytic stages of P. falciparum development. Based on the cytosolic localization of both PFB0595w and PfHsp70-1, a chaperone – cochaperone partnership was proposed that formed the basis for the in vitro experiments. PFB0595w was shown for the first time to stimulate the ATPase activity of PfHsp70-1 pointing to a functional interaction. Preliminary surface plasmon spectroscopy analysis has revealed a potential interaction between PFB0595w and PfHsp70-1 but highlights the need for further related experiments to support the findings. Gel filtration analysis showed that PFB0595w exists as a dimer thereby confirming in silico predictions. Based on these observations, we conclude that PFB0595w may regulate the chaperone activity of PfHsp70-1 in the cytosol while Pfj1 may play a co-chaperoning role for PfHsp70-3 in the mitochondrion. Overall, this data is expected to increase the knowledge of the Hsp70-J protein partnerships in the erythrocytic stage of P. falciparum development, thereby enhancing the understanding of parasite biology.

Este estudo objetivou estudar os carrapatos e a infecção por patógenos, e sua correlação com os vertebrados desta região entre 10/2007 e 09/2008 foram realizadas mensalmente doze visitas a uma área de floresta ombrófila densa da Amazônica na Estação Ecológica de Samuel. Foram coletados, 60 aves, 36 mamíferos, 14 répteis e 11 anfíbios. Em vida livre foram realizadas 80 coletas de carrapatos em um total de 39.600m em cerca de 2.640min (44h) e coletados 265 carrapatos adultos de quatro espécies, sendo elas: A. scalpturatum (81), A. latepunctatum (84), A. oblongogutattum (18), A. naponense (69) e 597 ninfas de Amblyomma sp. Dos 104 adultos (11,8% do total de carrapatos) foram testados para Anaplasmataceae, obtive-se 27 (25,9%) positivos e mais 36 adultos (total de 140) e 11 carrapatos foram positivos (1) A. scalpturatum, (4) A. latepunctatum, (3) A. naponense. É notável o impacto da hidrelétrica sobre a fauna de pequenos mamíferos. / This study was aimed to analyze the ticks and the pathogenic infections, and their correlation with the vertebrates of such a region from October 2007 to September 2008, 12 surveys were monthly carried out at the Samuel Ecological Station in an Amazonian dense ombrophilous forest. They were collected from 60 birds, 36 mammals, 14 reptiles, and 11 amphibians. Eighty(80) collections of free-living ticks were carried out within an area of about 39.000 meters, during 2.640 minutes (44 hours), and 265 adult ticks or four species were picked up as following: 81 A.scalpturatum, 84 A. latepunctatum, 18 A. oblongogutattum, 69 A. naponense , and 597 Amblyomma sp. nymphs. 104 the adults, which accounted for 11.8% of total of ticks collected, were tested for Anaplasmataceae, and 27 (25.9%) were positive. In the Rickettsia spp. sample, additionally to the above mentioned 104 ticks tested, 36 adult ticks (out of a total of 140) were positive. It is noteworthy the impact caused by the hydropower plant on the small mammal fauna.