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The common coral trout Plectropomus leopardus (Lacepède, 1802) is an important commercially and recreationally fished species in Australia. This thesis examined a number of factors that may affect the suitability of this species for aquaculture including ectoparasite fauna and stress physiology. Examination of the ectoparasite fauna of P. leopardus revealed one new species, Hatschekia (Species A), and new host records for the copepods Peniculisa bellwoodi and Sagum epinepheli. Other ectoparasites recorded included Dissonus manteri, Hatschekia plectropomi, Lepeophtheirus plectropomi, Dentigryps litus, Trochopus plectropomi, Diplectanum plectropomi, Aega lethrina, Argathona macronema and turbellarians. There were significant differences between the ectoparasite communities of P. leopardus from Heron Island Reef and a population from the Cairns region of the Great Barrier Reef with absence of some parasites from certain sites and variations in the intensity of infection of other parasites. Dissonus manteri, H. plectropomi and P. bellwoodi were associated with the most severe pathology as determined by gross examination, histology, scanning electron microscopy and vascular casting techniques. This pathology was considered unlikely to cause a significant threat to host fish survival at the infection intensities observed on wild fish. This hypothesis was supported by the observation that levels of ectoparasite infection of wild fish were insufficient to initiate a stress response, as determined by blood parameter analysis. Detailed examination of the distribution of H. plectropomi on the gills of P. leopardus revealed a site preference for holobranchs 2 and 3. Filaments near the ends of gill arches and those around the bend in the gill arch were rarely occupied. The chance of coming into contact with a suitable attachment site along with the ability to withstand ventilation forces at that site may be the major factors affecting the distribution of this species. Two H. plectropomi morphotypes were identified based on the direction of body curvature. Regardless of morphotype, 98% of individuals were attached such that the convex side of the body was oriented towards the oncoming ventilating water currents. Further, 94% of individuals attached to the posterior faces of filaments leading to a predictable pattern of attachment for this species. Baseline concentrations of 12 blood parameters were determined for 30 fish species representing 12 families from Heron Island Reef, Great Barrier Reef, including P. leopardus (Serranidae), in addition to one estuarine species, the fork-tailed catfish Arius graeffei Kner & Steindachner, 1867 (Ariidae), from the Brisbane River. Baseline physiological status was determined via rapid blood sampling via caudal puncture of wild caught fish and analysis of a range of blood parameters. The validity of this method was evaluated via analysis of blood samples taken from hussar Lutjanus adetii (Castelnau, 1873). There were no significant changes in any of the measured blood parameters during the first two minutes following a stressful stimulus supporting the use of rapid blood sampling for baseline blood parameter determination. The measured baseline values provided a basis for comparison of post-stress values with differences between species in baseline and post-stress physiological condition discussed in relation to differences in lifestyle such as activity level and diet. Examination of the response of coral trout to hook-and-line capture over a 3-day period revealed an initial elevation of cortisol from a median baseline concentration of 0.40 nM (n = 17) to a peak of 19.01 nM (n = 7) 2 h post-hooking. Plasma osmolality, glucose and lactate also rose significantly and all parameters had returned to near baseline levels within 24 h of capture. Cortisol, glucose and lactate were identified as the best indicators of acute physiological disturbance in P. leopardus. Cortisol, glucose, lactate, osmolality and haematocrit were the best indicators of acute physiological stress in A. graeffei. An acute temperature increase of approximately 5oC over a period of 15 minutes was stressful to P. leopardus as indicated by an elevation of plasma cortisol. In contrast, an acute temperature drop did not have any significant effect on the physiological parameters measured. In an acute handling and air exposure experiment fish were rapidly hand-netted from holding tanks, subjected to a period of air exposure (0, 2, 4 or 6 minutes) and returned to the water for two hours prior to being blood sampled. Air exposure time did not significantly affect the resultant physiological parameters, suggesting that netting and transfer was the most stressful part of the experimental protocol. The potential commercial applications of these findings are discussed.

The poultry red mite Dermanyssus gallinae (De Geer) is a blood feeding ectoparasite that infests many bird species. Economically it is the most important parasite affecting egg-laying hens. The aim of this study was to identify vaccine candidate proteins from D. gallinae using a number of approaches. An immunisation trial was conducted using four protein fractions derived from D. gallinae. The fractions were injected into hens and immunoglobulin Y was purified from the yolk of eggs laid by the hens. An in vitro feeding assay in which mites were fed these antibodies showed a significant increase (p = 0.013) in mortality of mites fed with antibodies against a PBS-soluble fraction of D. gallinae. The PBS-soluble proteins were then fractionated by anion exchange chromatography into three fractions, which were then used to immunise hens. One fraction produced antibodies that caused the greatest percentage mortality of mites, based on the results from a single feeding assay. To identify further potential vaccine antigens, three expressed sequence tag (EST) databases were produced. A complementary DNA (cDNA) library was prepared from a mixed gender and life stage population and 46 contigs were identified with significant homology to proteins from other organisms. A second database was generated using a suppression subtractive hybridisation approach and 133 “fed mite enriched” contigs identified with significant homology to proteins in either the NCBI non-redundant (nr) protein, KEGG databases or had a domain predicted by InterProScan. The most abundant proteins in this database were vitellogenin and GP80, a precursor molecule of vitellogenin. Roche 454 sequencing technology was used to generate an EST database of 13,363 contigs, of which 26 % had significant homology to a protein in the NCBI nr protein database. The majority of contigs (45 %) were classified as Cellular Processes and Signalling proteins. Illumina Solexa sequencing technology was also used to analyse the expression levels of genes in fed and starved mites. A total of 66 contigs were obtained with a significant and greater than three-fold change in expression level between the two groups. The contig with the largest fold change was homologous to vitellogenin (fold change 110). Paramyosin and tropomyosin have been used successfully to immunise hosts against other parasitic species. Tropomyosin had previously been characterised in D. gallinae. In this study paramyosin was characterised and recombinant versions of both proteins were used in an immunisation trial. Anti-tropomyosin (p < 0.001) and anti-paramyosin (p = 0.004) antibodies fed to mites in the in vitro feeding assay caused a significant increase in the mortality of the mites when compared to controls. An in vivo challenge was performed injecting three groups of hens with the PBS-soluble native protein fraction, a recombinant protein cocktail, consisting of paramyosin, tropomyosin, cathepsin L, cathepsin D and histamine release factor, and an adjuvant only control, which were subjected to a challenge of D. gallinae. Each group consisted of three replicates based on weight. Western blot analysis of hen serum showed a significant increase (p < 0.05) in the titre of antibody from the hens immunised with the recombinant cocktail compared with controls. One of the replicates immunised with the PBS-soluble protein fraction showed a strong response but this was lacking in the other two replicates and no significant difference in IgY titre was found. No significant differences were found between the number of mites collected from the test or control groups following a large mite challenge. In conclusion the PBS-soluble protein fraction, paramyosin and tropomyosin have been shown to have potential as vaccine candidates based on in vitro studies. The bioinformatic analysis of D. gallinae has provided a large EST database and a list of 66 proteins that had a significant difference in expression levels in fed and starved mites, which can be further mined for potential vaccine candidates.

No description available.

Made available in DSpace on 2014-06-11T19:27:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-07-22Bitstream added on 2014-06-13T19:36:20Z : No. of bitstreams: 1 correa_rr_me_jabo.pdf: 736045 bytes, checksum: 5d27ca5fc145e51fb8260e5449848591 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Uma nova associação ectoparasiticida contendo cipermetrina 15% + clorpirifós 30% + fenthion 15%, administrada via tópica “pulverização”, foi avaliada em cinco experimentos para determinar a eficácia terapêutica e segurança clínica. A atividade carrapaticida foi avaliada em dois ensaios, utilizando bovinos natural e artificialmente infestados pelo Rhipicephalus (Boophilus) microplus. Outros dois ensaios foram realizados para avaliar a eficácia da formulação contra Haematobia irritans e larvas de Dermatobia hominis em bovinos naturalmente infestados. A segurança clínica da associação foi aferida por exames clínicos, hematológicos e bioquímicos. A associação alcançou eficácia carrapaticida (infestação natural) superior a 95% no 7º, 14º e 21º DPT. A eficácia máxima alcançada foi de 97,38% no 14º DPT, declinando no 28º DPT para 74,97%. As quantificações de partenóginas de R. (B.) microplus no grupo medicado foram significativamente (P< 0,05) inferiores às do grupo controle, do 1º ao 28o DPT. Nos bovinos artificialmente infestados, verificou-se que a formulação avaliada atingiu eficácia superior a 90% do 3º ao 36º dia pós-tratamento (DPT). Do 5º ao 33º DPT, os índices de eficácia anti-ixodídica foram superiores a 95%. O número de teleóginas desprendidas do grupo tratado foi estatisticamente (P<0,05) inferior ao registrado no grupo controle ao longo de todo experimento. Contra H. irritans a associação supracitada alcançou eficácia mosquicida superior a 99% no 1o e 3o dias pós-tratamento (DPT). No 7o e 14º DPT, a associação alcançou eficácia mosquicida de 82,57% e 60,10%, respectivamente. As contagens de H. irritans realizadas nos bovinos pertencentes ao grupo tratado foram estatisticamente inferiores (P<0,05) às do grupo controle, do 1o ao 14o DPT. Contra larvas de Dermatobia hominis, a formulação apresentou eficácia... / A new association ectoparasiticide containing cypermethrin 15% + chlorpyrifos 30% + fenthion 15%, administered by topical spray, was evaluated in five experiments to determine the therapeutic efficacy and clinical safety. The insecticide activity was evaluated in two essays, using natural and artificially infested bovine by Rhipicephalus (Boophilus) microplus. Two other essays were conducted to evaluate the effectiveness of Haematobia irritans and the formulation against larvae of Dermatobia hominis in bovine naturally infested. The clinical safety of the association was compared by clinical, haematological and biochemistry tests. The association achieved effectiveness insecticide (natural infestation) exceeding 95% in the 7th, 14th and 21st DPT. Maximum effectiveness achieved was 97.38% in 14th DPT, declining in 28th DPT to 74.97%. The quantifications of R. (B.) microplus medicated group were significantly (P 0.05 <) lower than the control group, from 1st to 28th DPT. In bovine artificially infested, it was found that the formulation evaluated reached effectiveness exceeding 90% of 3rd to 36th day post-treatment (DPT). From 5th to 33th DPT, the indexes of efficiency were anti-ixodídica exceeding 95%. The number of R. (B.) microplus loosed of the treated group was statistically (P<0.05) less than that recorded in the tracking group throughout the experiment. Against H. irritans the abovementioned association achieved effectiveness exceeding 99% in the 1st and 3rd days post-treatment (DPT). In the 7th and 14th DPT, membership reached 82.57% effectiveness and insecticide of 60.10% respectively. The counts of H. irritans performed in bovine belonging to the treated group were statistically lower (P<0.05) to the control group, from the 1st to the 14th DPT. Against larvae of Dermatobia hominis, the formulation presented effectiveness of 100%, in the 7th DPT. A statistically significant ... (Complete abstract click electronic access below)

No description available.

Ce travail s’est intéressé aux liens existant entre la stratégie d’exploitation développée par un crustacé ectoparasite et son comportement reproductif. Le crabe Pinnotheridae Dissodactylus primitivus exploite deux espèces Spatangidae vivant dans la Mer des Caraïbes, Meoma ventricosa et Plagiobrissus grandis. Des approches comportementales, démographiques et génétiques ont été adoptées afin de mettre en lumière le fonctionnement et la biologie de cette symbiose. Par son comportement alimentaire, le crabe occasionne des lésions tégumentaires sur ses hôtes. Celles-ci affectent la fitness de M. ventricosa, au travers de son développement gonadique. Dissodactylus primitivus exploite ses deux espèces hôtes de façon asymétrique. La reproduction des parasites se déroule sur les deux hôtes, alors que le recrutement ne s’effectue que sur M. ventricosa. Ce cycle vital asymétrique du crabe serait stabilisé par la qualité et la rareté de P. grandis. En outre, Le comportement sexuel du crabe sur M. ventricosa répondrait aux critères de la polygynandrie à femelles mobiles. Selon ce modèle, les mâles et les femelles se déplacent entre les hôtes à la recherche de partenaires multiples. Lors de ces déplacements, le crabe s’aiderait de son aptitude à localiser chimiquement ses hôtes. Néanmoins, ce mécanisme s’avère plastique et pourrait refléter l’asymétrie du cycle vital. En effet, cette différence n’a pas d’origine génétique, car les crabes vivant au sein du site d’étude constituent la même population quelle que soit l’espèce hôte considérée. Les marqueurs moléculaires microsatellites mis au point dans ce travail permettront lors de futurs travaux d’affiner les observations sur les modalités d’accouplement du crabe et d’estimer sa capacité de dispersion. / This work aimed to highlight the relationships between the host exploitation strategy of an ectoparasite crustacean and its mating system. The pea crab Dissodactylus primitivus exploits two Spatangidae species living in the Caribbean Sea, Meoma ventricosa and Plagiobrissus grandis. Behavioural, demographic and genetic approaches have been conducted to examine the functioning and biology of this symbiosis. Owing to its feeding behaviour, the crab wounds the host tegument. The wounds negatively affect M. ventricosa's fitness through its gonadic development. Dissodactylus primitivus asymmetrically exploits its two host species. The reproduction of the parasites happens on each host, but the recruitment only takes place on M. ventricosa. The asymmetrical life cycle would be stabilised par the quality and the scarcity of P. grandis. The mating system of crabs living on M. ventricosa would correspond to the Pure-search polygynandry of mobile females criteria. According to this model, the males and the females practice the host switching behaviour to find several sexual partners. During these movements, the crab could use its chemodetection ability to locate its hosts. However, this mechanism is plastic and presumably reflects the asymmetrical life cycle of the crab. This difference has indeed not a genetic cause because the crabs living inside the investigated region belong to the same population, whatever the regarded host species. In future studies, the microsatellites markers developed for this work could be used to test the mating system of D. primitivus and to estimate its dispersion ability.

Orientador: Alvimar José da Costa / Banca: Luciano Melo de Souza / Banca: Gilson Pereira de Oliveira / Resumo: Uma nova associação ectoparasiticida contendo cipermetrina 15% + clorpirifós 30% + fenthion 15%, administrada via tópica "pulverização", foi avaliada em cinco experimentos para determinar a eficácia terapêutica e segurança clínica. A atividade carrapaticida foi avaliada em dois ensaios, utilizando bovinos natural e artificialmente infestados pelo Rhipicephalus (Boophilus) microplus. Outros dois ensaios foram realizados para avaliar a eficácia da formulação contra Haematobia irritans e larvas de Dermatobia hominis em bovinos naturalmente infestados. A segurança clínica da associação foi aferida por exames clínicos, hematológicos e bioquímicos. A associação alcançou eficácia carrapaticida (infestação natural) superior a 95% no 7º, 14º e 21º DPT. A eficácia máxima alcançada foi de 97,38% no 14º DPT, declinando no 28º DPT para 74,97%. As quantificações de partenóginas de R. (B.) microplus no grupo medicado foram significativamente (P< 0,05) inferiores às do grupo controle, do 1º ao 28o DPT. Nos bovinos artificialmente infestados, verificou-se que a formulação avaliada atingiu eficácia superior a 90% do 3º ao 36º dia pós-tratamento (DPT). Do 5º ao 33º DPT, os índices de eficácia anti-ixodídica foram superiores a 95%. O número de teleóginas desprendidas do grupo tratado foi estatisticamente (P<0,05) inferior ao registrado no grupo controle ao longo de todo experimento. Contra H. irritans a associação supracitada alcançou eficácia mosquicida superior a 99% no 1o e 3o dias pós-tratamento (DPT). No 7o e 14º DPT, a associação alcançou eficácia mosquicida de 82,57% e 60,10%, respectivamente. As contagens de H. irritans realizadas nos bovinos pertencentes ao grupo tratado foram estatisticamente inferiores (P<0,05) às do grupo controle, do 1o ao 14o DPT. Contra larvas de Dermatobia hominis, a formulação apresentou eficácia ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: A new association ectoparasiticide containing cypermethrin 15% + chlorpyrifos 30% + fenthion 15%, administered by topical "spray", was evaluated in five experiments to determine the therapeutic efficacy and clinical safety. The insecticide activity was evaluated in two essays, using natural and artificially infested bovine by Rhipicephalus (Boophilus) microplus. Two other essays were conducted to evaluate the effectiveness of Haematobia irritans and the formulation against larvae of Dermatobia hominis in bovine naturally infested. The clinical safety of the association was compared by clinical, haematological and biochemistry tests. The association achieved effectiveness insecticide (natural infestation) exceeding 95% in the 7th, 14th and 21st DPT. Maximum effectiveness achieved was 97.38% in 14th DPT, declining in 28th DPT to 74.97%. The quantifications of R. (B.) microplus medicated group were significantly (P 0.05 <) lower than the control group, from 1st to 28th DPT. In bovine artificially infested, it was found that the formulation evaluated reached effectiveness exceeding 90% of 3rd to 36th day post-treatment (DPT). From 5th to 33th DPT, the indexes of efficiency were anti-ixodídica exceeding 95%. The number of R. (B.) microplus loosed of the treated group was statistically (P<0.05) less than that recorded in the tracking group throughout the experiment. Against H. irritans the abovementioned association achieved effectiveness exceeding 99% in the 1st and 3rd days post-treatment (DPT). In the 7th and 14th DPT, membership reached 82.57% effectiveness and insecticide of 60.10% respectively. The counts of H. irritans performed in bovine belonging to the treated group were statistically lower (P<0.05) to the control group, from the 1st to the 14th DPT. Against larvae of Dermatobia hominis, the formulation presented effectiveness of 100%, in the 7th DPT. A statistically significant ... (Complete abstract click electronic access below) / Mestre

Ce travail s’est intéressé aux liens existant entre la stratégie d’exploitation développée par un crustacé ectoparasite et son comportement reproductif. Le crabe Pinnotheridae Dissodactylus primitivus exploite deux espèces Spatangidae vivant dans la Mer des Caraïbes, Meoma ventricosa et Plagiobrissus grandis. Des approches comportementales, démographiques et génétiques ont été adoptées afin de mettre en lumière le fonctionnement et la biologie de cette symbiose. Par son comportement alimentaire, le crabe occasionne des lésions tégumentaires sur ses hôtes. Celles-ci affectent la fitness de M. ventricosa, au travers de son développement gonadique. Dissodactylus primitivus exploite ses deux espèces hôtes de façon asymétrique. La reproduction des parasites se déroule sur les deux hôtes, alors que le recrutement ne s’effectue que sur M. ventricosa. Ce cycle vital asymétrique du crabe serait stabilisé par la qualité et la rareté de P. grandis. En outre, Le comportement sexuel du crabe sur M. ventricosa répondrait aux critères de la polygynandrie à femelles mobiles. Selon ce modèle, les mâles et les femelles se déplacent entre les hôtes à la recherche de partenaires multiples. Lors de ces déplacements, le crabe s’aiderait de son aptitude à localiser chimiquement ses hôtes. Néanmoins, ce mécanisme s’avère plastique et pourrait refléter l’asymétrie du cycle vital. En effet, cette différence n’a pas d’origine génétique, car les crabes vivant au sein du site d’étude constituent la même population quelle que soit l’espèce hôte considérée. Les marqueurs moléculaires microsatellites mis au point dans ce travail permettront lors de futurs travaux d’affiner les observations sur les modalités d’accouplement du crabe et d’estimer sa capacité de dispersion. This work aimed to highlight the relationships between the host exploitation strategy of an ectoparasite crustacean and its mating system. The pea crab Dissodactylus primitivus exploits two Spatangidae species living in the Caribbean Sea, Meoma ventricosa and Plagiobrissus grandis. Behavioural, demographic and genetic approaches have been conducted to examine the functioning and biology of this symbiosis. Owing to its feeding behaviour, the crab wounds the host tegument. The wounds negatively affect M. ventricosa's fitness through its gonadic development. Dissodactylus primitivus asymmetrically exploits its two host species. The reproduction of the parasites happens on each host, but the recruitment only takes place on M. ventricosa. The asymmetrical life cycle would be stabilised par the quality and the scarcity of P. grandis. The mating system of crabs living on M. ventricosa would correspond to the Pure-search polygynandry of mobile females criteria. According to this model, the males and the females practice the host switching behaviour to find several sexual partners. During these movements, the crab could use its chemodetection ability to locate its hosts. However, this mechanism is plastic and presumably reflects the asymmetrical life cycle of the crab. This difference has indeed not a genetic cause because the crabs living inside the investigated region belong to the same population, whatever the regarded host species. In future studies, the microsatellites markers developed for this work could be used to test the mating system of D. primitivus and to estimate its dispersion ability.

The aim of the thesis was to verify use of peracetic acid (PAA) in production conditions for ectoparasites control in carp pond farming. First test was about evaluation of organic load in pond water on speed PAA decay in two temperature regime (11 15 and 21 °C). Results showed that between 3 6 hours from PAA application the concentration of PAA was under a half of the initial concentration (6 mg.l-1). Also statically important differences (p= < 0.05) were found in speed of decay between samples with organic load (3 samples from ponds) and samples without organic load (distilled and tap water) in both temperature regimes. Application of PAA during the transport of fish into the transport boxes was performed in three age categories (1, 2 and 3 years old common carps), where the fish were exposed for 60 90 minutes in 1.5 mg.l-1 PAA as Persteril 36. Parasitic examinations of skin and gills showed reduction of number protozoal parasites (Trichodina spp. and Apiosoma spp.) in individuals comparing with control. Against other detected parasites was PAA ineffective. Statistical evaluation showed significant (p= <0.05) decrease in prevalence only in 3 cases out of 19 in total. Long-term four days long fish bath of carps which were heavily infested by Saprolegnia parasitica in 1.5 mg.l-1 PAA an positive effect on the overall survival was found. In the control group mortality reached 100 % the third day of trial, in contrast with treated group where was Saprolegnia parasitica succesfuly eliminated and overall moratily was 40 %. Continual fourteen days long application of PAA by peristaltic pump did not proved.