Dissertations / Theses on the topic 'Animal Parasitic Diseases'

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Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq
The herbal drugs have been widely studied in order to obtain an effective and ecologically viable alternative for the control of parasitic diseases of domestic animals. It was evaluated the spectrum of antiparasitic action of the seed oil of Carapa guianensis against muscidae, ticks, lice and gastrointestinal nematodes of domestic animals. The action against muscidae was assessed against the first larval (L1) and third stage (L3) from Musca domestica and Stomoxys calcitrans adults. For ticks it was determined the IC50 (Inhibitory Concentration Medium) and LC50 (median lethal concentration) on Rhipicephalus (Boophilus) microplus, and Rhipicephalus sanguineus Anocentor nitens. Against lice it was determined the efficacy against Damalinia caprae. The in vivo efficacy in mice naturally infected with Syphacia obvelata and in goats and sheep naturally infected with gastrointestinal helminths was determined and also the effect of oral administration of seed oil of C. guianensis on hematological and biochemical parameters in goats and sheep. The results revealed that the seed oil of Carapa guianensis exerted negative effect on M. domestica bioactivity causing severe mortality and reduced larval and pupal weight of larvae, show repellency on Stomoxys calcitrans and have significant potential in the control of ticks of domestic animals, interfering with their reproduction revealing an IC50 of 4.332, 4.850, 4.903, and an LC50 of 5.228, 5.362 and 5.698, respectively, for females and larvae of R. (B.) microplus, A. nitens and Rhipicephalus sanguineus. The seed oil of Carapa guianensis has in vitro activity against adult Damalinia caprae and the anthelmintic tests using mice, goats and sheep revealed an ineffective anthelmintic activity in all treatments. The evaluation of hematological and biochemical parameters of goats and sheep in general did not determine difference in the values of the erythrocyte, total leukocyte and biochemical parameters.
Os fitoterápicos têm sido amplamente estudados visando-se obter uma alternativa eficaz e ecologicamente viável para o controle das doenças parasitárias dos animais domésticos. Avaliou-se o espectro de ação antiparasitária do óleo da semente de Carapa guianensis, Aubl. contra muscídeos, ixodídeos, fitirápteros e nematóides gastrintestinais de animais domésticos. A ação contra muscídeos foi avaliada contra larvas de primeiro (L1) e terceiro estádio (L3) de Musca domestica e adultos de Stomoxys calcitrans. Para os carrapatos determinou-se a CI50 (Concentração Inibitória Média) e CL50 (Concentração Letal Média) sobre Rhipicephalus (Boophilus) microplus, Anocentor nitens e Rhipicephalus sanguineus. Contra os fitirápteros determinou-se a eficácia contra Damalinia caprae. Com relação aos helmintos gastrintestinais determinou-se in vivo a eficácia em infectados com o oxiurídeo Syphacia obvelata e contra nematóides gastrintestinais de caprinos e ovinos naturalmente infectados. Foi avaliado ainda o efeito da administração oral do óleo da semente da C. guianensis sobre os parâmetros hematológicos e bioquímicos em caprinos e ovinos. Os resultados revelaram que o óleo da semente da Carapa guianensis exerce bioatividade negativa sobre M. domestica, provocando acentuada mortalidade larval e pupal e redução no peso das pupas; apresenta atividade de repelência sobre Stomoxys calcitrans; possui significativo potencial no controle dos ixodídeos dos animais domésticos, interferindo na sua reprodução revelando uma CI50 de 4,332; 4,850; 4,903, e uma CL50 de 5,228; 5,362 e 5,698, respectivamente, para fêmeas e larvas de R. (B.) microplus, A. nitens e Rhipicephalus sanguineus; apresenta atividade piolhicida in vitro contra Damalinia caprae adultos. Os testes antihelmínticos utilizando como modelo experimental camundongos revelaram atividade antihelmíntica não efetiva em todos os tratamentos utilizados com eficácia variando de 30,36% a 54,72%; quando administrado por via oral em caprinos e ovinos naturalmente infectados por nematóides gastrintestinais não se apresentou efetivo na redução da eliminação de ovos e na inibição do desenvolvimento de larvas de nematóides gastrintestinais. A avaliação dos parâmetros hematológicos e bioquímicos dos caprinos e ovinos de um modo geral não determinou diferença significativa nos valores do eritrograma, leucócitos totais e parâmetros bioquímicos.

Vaccines and anthelmintics induce an inflammatory response in equids. Since they are commonly given concurrently, it is practical to study any interaction between them. This study evaluated whether IVM and PYR would modulate the acute phase inflammatory response, the systemic gene expression of pro-inflammatory cytokines, and vaccine-specific titers induced by WNV, EHV, and KLH vaccines. Naturally-infected, yearling ponies were sorted by gender, then fecal epgs. They were randomly assigned to three treatment groups: IVM, PYR, and control. All ponies received vaccinations intramuscularly on days 0 and 29. Whole blood, serum, and plasma samples were collected 1, 3, and 14 days post-vaccination. Samples were analyzed for inflammatory markers, cytokine, mRNA expression, and vaccine-specific IgG titers by ELISA. The acute-phase inflammatory marker data showed no statistical significance; they did show an increase in SAA, haptoglobin, and fibrinogen, and a decrease in iron after vaccination. The mRNA data showed that anthelmintics had a significant effect on interleukin mRNA levels, but not on TNF-α or IFN-γ levels. The ELISA assays showed no biologically significant reduction in IgG as compared to the control group. We conclude that deworming does not affect vaccine IgG titers; therefore, ceasing vaccinating and deworming concurrently is not necessary.

Streptococcus agalactiae is a pathogen of multiple hosts. The bacterium, an aetiological agent of septicaemia and meningo-encephalitis in freshwater and saltwater fish species, is considered a major threat to the aquaculture industry, particularly for tilapia. Cattle and humans are however the main known reservoirs for S. agalactiae. In humans, the bacterium (commonly referred to as Group B Streptococcus or GBS) is a member of the commensal microflora of the intestinal and genito-urinary tracts, but it is also a major cause of neonatal invasive disease and an emerging pathogen in adults. In cattle, S. agalactiae is a well-recognized causative agent of mastitis. Numerous studies focusing on S. agalactiae from human and bovine origins have provided insight into the population structure of the bacterium, as well as the genome content and pathogenic mechanisms through identification of virulence determinants. Concerning S. agalactiae from aquatic origins, scientific information mainly focused on case reporting and/or experimental challenges, with a limited or absence of information in terms of pathogenesis, virulence determinants and genotypes of the strains involved. The objective of this study was to enhance our understanding of the molecular epidemiology, host-adaptation and pathogenicity of S. agalactiae in aquatic species, with particular emphasis on tilapia. Firstly, a collection of 33 piscine, amphibian and sea mammal isolates originating from several countries and continents was assembled, with the aim of exploring the population structure and potential host specificity of aquatic S. agalactiae. Isolates were characterised using pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), and a standardised 3-set genotyping system comprising molecular serotypes, surface protein gene profiles and mobile genetic element profiles. Two major subpopulations were identified in fish. The first subpopulation consisted of non-haemolytic isolates that belonged to sequence type (ST) 260 or 261, which are STs that have been reported only from teleosts. These isolates exhibited a low level of genetic diversity by PFGE and clustered with other STs that have been reported only in fish. Another common feature was the absence of all surface protein genes or mobile genetic elements targeted as part of the 3-set genotyping and that are usually found in human or bovine isolates. The second subpopulation consisted of β-haemolytic isolates recovered from fish, frogs and sea mammals, and that exhibited medium to high genetic diversity by PFGE. STs identified among these isolates have previously been identified from strains associated with asymptomatic carriage and invasive disease in humans. The human pathogenic strain ST7 serotype Ia was detected in fish from Asia. Moreover, ST283 serotype III-4 and its novel single locus variant ST491 detected in fish from Southeast Asia shared a 3-set genotype identical to that of an emerging ST283 clone associated with invasive disease of adult humans in Asia. These observations suggested that some strains of aquatic S. agalactiae may present a zoonotic or anthroponotic hazard. STs found among the seal isolates (ST23) have also been reported from humans and numerous other host species, but never from teleosts. This work provided an excellent basis for exploration of the virulence of selected strains in experimental challenges. The virulence of two strains of S. agalactiae was experimentally investigated by intra-peritoneal infection of Nile tilapia (Oreochromis niloticus), using an isolate originally recovered from fish and belonging to ST260, and an isolate originating from a grey seal and belonging to ST23. The clinical signs, the in vivo distribution of viable bacteria and bacterial antigens, and the gross and histopathological lesions that developed during the time course of the infection were investigated. The ST260 strain was highly virulent, whereas no major clinical sign or mortalities occurred in the fish challenged with the ST23 strain. After injection, both strains however gained access to the bloodstream and viable bacteria were recovered from all organs under investigation. During the early stages of infection, bacteria were mostly found within the reticulo-endothelial system of the spleen and kidney. Thereafter, the ST260 demonstrated a particular tropism for the brain and the heart, but granulomatous inflammation and associated necrotic lesions were observed in all organs. ST23 was responsible for a mixed inflammatory response associated with the presence of bacteria in the choroid rete and in the pancreatic tissue only. After 7 days post-challenge and for both strain, the formation or containment of bacteria within granulomata or other encapsulated structures appeared to be a major component of the fish response. However, the load of viable bacteria remained high within organs of fish infected with ST260, suggesting that, unlike ST23, this strain is able to survive within macrophages and/or to evade the immune system of the fish. This work demonstrates that the lack of report of ST23 strains in fish is possibly not due to a lack of exposure but to a lack of virulence in this host. The two strains, which differ in prevalence and virulence in fish, provide an excellent basis to investigate genomic differences underlying the host-association of distinct S. agalactiae subpopulations. The genome of the ST260 strain used in challenge studies was sequenced. We therefore provided the first description for the genome sequence of a non-haemolytic S. agalactiae isolated from tilapia (strain STIR-CD-17) and that belongs by multi-locus sequence typing (MLST) to clonal complex (CC) 552, which corresponds to a presumptive fish-adapted subgroup of S. agalactiae. The genome was compared to 13 S. agalactiae genomes of human (n=7), bovine (n=2), fish (n=3) and unknown (n=1) origins. Phylogenetic analysis based on the core genome identified isolates of CC552 as the most diverged of all S. agalactiae studied. Conversely, genomes from β-haemolytic isolates of CC7 recovered from fish were found to cluster with human isolates of CC7, further supporting the possibility that some strains may represent a zoonotic or anthroponotic hazard. Comparative analysis of the accessory genome enabled the identification of a cluster of genes uniquely shared between CC7 and CC552, which encode proteins that may provide enhanced fitness in specific niches. Other genes identified were specific to STIR-CD-17 or to CC552 based on genomic comparisons; however the extension of this analysis through the PCR screening of a larger population of S. agalactiae suggested that some of these genes may occasionally be present in isolates belonging to CC7. Some of these genes, occurring in clusters, exhibited typical signatures of mobile genetic elements, suggesting their acquisition through horizontal gene transfer. It is not possible to date to determine whether these genes were acquired through intraspecies transfer or through interspecies transfer from the aquatic environment. Finally, general features of STIR-CD-17 highlighted a distinctive genome characterised by an absence of well conserved insertion sequences, an abundance of pseudogenes, a smaller genomic size than normally observed among human or bovine S. agalactiae, and an apparent loss of metabolic functions considered conserved within the bacterial species, indicating that the fish-adapted subgroup of isolates (CC552) has undergone niche restriction. Finally, genes encoding recognised virulence factors in human S. agalactiae were selected and their presence and structural conservation was evaluated within the genome of STIR-CD-17.

The dusky grouper Epinephelus marginatus (Lowe 1834), is a protogynous sequential hermaphrodite and is considered to be one of the most important fish species in the Mediterranean Sea. It is a K-strategist, being slow growing and late maturing, and this, coupled with its reproductive biology and relatively sedentary behaviour, has made it extremely sensitive to overexploitation, leading it to be classified by the IUCN as an endangered fish species. Wild dusky grouper have suffered from disease outbreaks in the past decade, leading to mass mortalities across the Mediterranean Sea, including Libyan coastal waters. These mortalities have mostly been attributed to Nodavirus infections. In Europe and Brazil, efforts are in place to culture this fish for commercial grow-out and stock enhancement programmes. In Libya, the dusky grouper is consumed regularly and is considered a prime-eating fish. Its importance for the Libyan internal market, as well as its potential for export, makes it an ideal candidate for future Libyan aquaculture activities. Given the scarce literature regarding the dusky grouper in Libya, this study aimed first to assess dusky grouper fisheries, spawning seasons and to identify the main threats that the fishing sector poses for wild stocks. Second this study aimed to determine the health status of wild dusky grouper offered at a local fish market in the capital Tripoli, in order to identify pathogens, pathologies or other health issues that might pose a hazard to cultured populations but also to remaining wild dusky grouper stocks. To achieve these aims, twelve field surveys spanning the period of 2013-2015 were conducted. From these surveys, it was established that the dusky grouper is captured throughout the year, including the spawning season. Fish sizes offered for sale ranged between 20-92 cm total length (TL), with the fish being sold from local fishing grounds around Tripoli, but also from as far as Benghazi, 1300 km to the east of Tripoli. The dusky grouper is principally caught in artisanal fisheries and by spearfishing, with approximately 300 spear-fishermen serving one particular fish market in Tripoli that was a focus in this study, and with dusky grouper being one of their main targets. Over the period of the survey, 267 landed dusky grouper were inspected for visible lesions prior to sampling. A total of 50 dusky grouper with sizes ranging from 27- 66 cm TL including the gonads from a further five fish measuring 66-92 cm TL that were sampled separately and examined to assess the stage of sexual maturity and to look for the presence of parasitic infections mainly affecting the gills, skin and gonads. The spawning season was found to extend from May to early September, with females ranging between 39-68 cm TL, males measuring 57-92 cm TL, and transient fish measuring 58-68 cm TL. From otolith readings of 8 fish, the youngest fish was a 3 year old juvenile of 28 cm TL and the oldest was an 8-9 year old 56 cm TL female. Whilst the highest prevalence of parasitic infection was found to be monogenean infection of the gills, with 100% prevalence, followed by gnathiid isopods infecting the oral cavity with 92% prevalence, it was the nematode Philometra sp. infecting post-spawning ovaries at 52% prevalence, that gave the highest apparent pathological impact. Necrosis potentially attributed to Philometra sp. in one particular ovary, was at a level likely to have caused complete parasitic castration, while others showed varying levels of probable functional reduction. The pathologies described need further investigation, especially in relation to possible synergies between Philometra sp. and bacteria in causing the necrosis. From the 267 inspected dusky grouper, 55 fish ranging in size from 42-92 cm TL were observed to be affected by external skin lesions of unknown aetiology. Twenty-six of these fish were sampled, having lesions at various stages of severity, and 5 further unaffected fish were used for histological assessment of the skin as negative controls. Histopathologically, the lesions comprised a multifocal, unilateral or bilateral dermatitis, involving the epidermis, superficial dermis and scale pockets, and sometimes, in severe cases, the hypodermis. Severe lesions had marked epidermal spongiosis progressing to ulceration. Healing was observed in some fish. Bacteria and fungi could be isolated from severe lesions, although they were not seen histopathologically in early-stage lesions. By contrast, metazoan parasite eggs were observed in the dermis and epidermis of some fish with mild and moderate dermatitis. Unidentified gravid digenean trematodes, carrying similar eggs, were also seen within the blood vessels of the deep and superficial dermis. The newly described condition was termed dusky grouper dermatitis (DGD). DGD’s geographical distribution along the Libyan coastline was investigated using a novel application of the social media network Facebook. Using Facebook, it was possible to document skin lesions of dusky grouper in Libyan waters from images attached to the entries of spear-fishermen. Thirty two Facebook accounts and 8 Facebook groups posting from 23 Libyan coastal cities provided a retrospective observational dataset comprising a total of 382 images of dusky grouper caught by spearfishing from December 2011-December 2015. Skin lesions were observable on 57 / 362 fish, for which images were of sufficient quality for analysis, giving a minimal prevalence for lesions of 15.75%. Only dusky grouper exceeding an estimated 40 cm total length exhibited lesions. The ability to collect useful data about the occurrence and geographical distribution of pathological conditions affecting wild fish using social media networks, demonstrates their potential utility as a tool to support epidemiological studies and monitor the health of populations of aquatic animals. The gravid digenean trematode described from mild lesions of five fish was identified using reconstruction through histological sectioning as belonging to the Family Aporocotylidae Odhner, 1912. This is the first description of a blood fluke from the dusky grouper, as well as from dermal blood vessels. The parasite was relatively long; the longest section of the parasite that could be measured was 1500 µm and 20-80 µm in width, while the total length of the parasite was estimated at 1500-2000 µm. Minute tegumental spines, possibly covering only a few parts of the parasite, were seen from some cross-sections. The parasite had one post-testicular ovary, which might overlap the testis, a pre-ovarian ascending uterus, and a post-ovarian descending uterus. It also possessed an oesophagus surrounded by oesophageal glandular cells and a pre-ovarian and pre-testicular extension of the vitelline cells, mostly at the level of the ascending uterus. The parasite was observed to be intra-vascular, the uterine lumen varies in size to accommodate between 1-7 eggs. The uterine eggs were embryonated and observed to span several stages of maturation. Eggs were also found in the dermal blood vessels, in the dermis, and in the epidermis, with the latter appearing to provide a potential route of egress of eggs into the environment. The extra-uterine eggs were 23.5 to 37.52 µm long and contained a ciliated miracidium. The eggs seemed to elicit a mixed inflammatory reaction, with degranulation of eosinophilic granular cells attached to the external surface of some of the eggs within the blood vessels but also the dermis. From observations made in the current study, this parasite appears to be a new species, most closely allied to none of the currently described Aporocotylidae genera.
In summary, the present study has demonstrated that the dusky grouper is extensively fished in Libya without discrimination to sizes and season, by both artisanal and spearfishing, with the latter as one of the main fishing methods, posing treats to the spawning potential and conservation of dusky grouper in Libya. The philometrid infecting the ovaries has a potential to reduce fecundity or to result in parasitic castration of wild broodstock. Gill-infecting monogeneans might represent a hazard for all stages of dusky grouper production. Dusky grouper dermatitis is a skin lesion, although there are no indications that infections may result in mortalities. Under culture conditions, however, this might change due to increase bacterial loads, which might lead to secondary bacterial infection. The presence of skin lesions would undoubtedly reduce the market value of whole fish. These findings are important for existing wild stocks, and for future plans regarding the aquaculture of dusky grouper. Future studies need to focus on the pathology of DGD, describing the disease process and aetiology using laboratory techniques such as TEM and virology as well as using morphology and molecular-based tools to describe the blood fluke and to determine their potential role in the initiation the disease. The novel approach to disease surveillance using social media Facebook posts could be further expanded by attracting citizen scientists, for future research assessing disease in wild fish, for sightings of mortality events and/or the appearance of disease outbreaks, or, for mapping marine mammal stranding’s and/or turtle nesting activity.

Les changements globaux, qu’ils soient d’origine climatique ou anthropique ont diverses conséquences en santé humaine et animale, mais aussi sur les écosystèmes mondiaux. L’une des plus importantes est la modification des aires de répartitions géographiques des espèces et de celle des pathogènes qui leurs sont associés. C’est dans ce contexte que nous assistons ces dernières années à une recrudescence des cas d’émergences et de réémergences de maladies infectieuses dans le monde. Alors que les efforts de recherche menés dans ce domaine se focalisent principalement sur les maladies virales, les maladies transmises par les mollusques d’eau douce, qui affectent plus d’un milliard d’individus dans le monde, sont également sujettes à ces évènements d’émergences devenus fréquents. Cependant, l’étude de la dynamique des parasites associés à ces maladies se focalisent essentiellement sur le diagnostic et le traitement des hôtes définitifs, en particulier l’Homme. Toutefois, une telle approche ne permet pas de prévenir de la transmission de ces parasites à l’Homme et encore moins de prévenir d’un évènement d’émergence, et les outils actuels utilisés pour le suivi de ces parasites dans l’environnement sont difficilement applicables à large échelle. Ce travail de thèse se propose donc d’apporter une vision plus environnementale de la dynamique de ces maladies. Avec l’exemple de l’émergence de bilharziose urogénitale en Corse, nous avons analysé cette émergence en intégrant l’étude des traits d’histoire de vie du parasite tropical en cause, notamment sa thermo tolérance, ainsi que le rôle des hôtes intermédiaires mollusques et des hôtes définitifs sauvages et domestiques dans le maintien local du cycle parasitaire. Dans un second temps nous avons développé des outils de diagnostic par ADN environnemental pour la détection de mollusques hôtes dans l’environnement afin d’identifier les zones à risque d’émergence, ainsi que des outils de détection intramolluscal de schistosomes pour identifier les sites de transmission actif, et donc permettre un suivi environnemental des acteurs de ces maladies. Pour compléter ces approches, nous avons développé un outil plus généraliste de metabarcoding environnemental pour caractériser les communautés de mollusques d’eau douce, et initié le développement d’un outil similaire pour la caractérisation des communautés de trématodes, ceci afin d’étudier les interactions entre ces organismes. Enfin nous discutons de l’intégrations de tous ces éléments dans de nouvelles stratégies de contrôle à l’encontre de maladies transmises par les mollusques d’eau douce
Global changes, whether climatic or anthropogenic, have various consequences in human and animal health, as well as for worldwide ecosystems. One of the most important is the modification of geographical ranges of species and those of their associated pathogens. It is in this context that in recent years we have witnessed a resurgence in the emergence and re-emergence of infectious diseases around the world. While research efforts in this field are mainly focused on viral diseases, freshwater snail-borne diseases, that affect more than 1 billion peoples around the world, are also subject to these outbreaks, which have become frequent. However, the study of the dynamics of parasites associated with these diseases focuses primarily on the diagnosis and treatment of the definitive hosts, particularly humans. Such an approach does not prevent the transmission of these parasites to humans and even less prevent an emergence event, and the existing tools used to monitor these parasites in the environment are difficult to apply at large scale. This thesis work, therefore aims to provide a more environmental vision of the dynamics of these diseases. With the example of the emergence of urogenital bilharziasis in Corsica, we analysed this emergence by integrating the study of the life history traits of the tropical parasite in question, particularly its thermo tolerance, as well as the role of mollusc intermediate hosts and wild and domestic definitive hosts in the local maintenance of the parasite lifecycle. In a second step, we have developed environmental DNA diagnostic tools for the detection of molluscs hosts in the environment in order to identify areas at risk of emergence, as well as tools for intramolluscal detection of schistosomes to identify active sites of transmission, and thus allow the environmental monitoring of the actors of these diseases. To complete these approaches, we have developed a more generalised environmental metabarcoding tool to characterise freshwater mollusc communities and initiated the development of a similar tool for the characterisation of trematode communities, in order to study the interactions between these organisms. Lastly, we discuss the integration of all these elements into new control strategies against snail-borne diseases

En permettant aux animaux de faire face à des environnements variables, l'apprentissage et la mémoire contribuent à l'optimisation de leur fitness, en leur permettant d'extraire et d'utiliser des informations, de façon à réduire l'incertitude associée à des environnements imprévisibles. Parmi les insectes, la drosophile et l'abeille domestique sont considérés comme des modèles classiques pour l'étude de l'apprentissage et de la mémoire. Les travaux réalisés sur ces derniers ont apporté une quantité considérable d'informations concernant les bases génétiques, neurobiologiques et moléculaires de ces processus, et ont permis de rendre compte du niveau de complexité des capacités cognitives des insectes. Cette somme de connaissances fondamentales acquises chez ces insectes contraste étonnamment avec le faible niveau de connaissance concernant la cognition des espèces impliquées dans des problématiques qui touchent la santé humaine et animale. Pourtant, il est largement admis que l'étude détaillée des capacités cognitives des insectes vecteurs de maladies constitue un aspect prioritaire pour la compréhension de leurs adaptations à la vie hématophage, de leur importance vectorielle, ainsi que pour le développement de nouveaux outils pour leur contrôle. Les travaux réalisés à ce jour chez les vecteurs, principalement chez les moustiques, ont été menés dans des contextes naturels ou peu contrôlés et ne proposent donc pas de démonstration formelle d'apprentissage. Le principal objectif de ce travail de thèse est de proposer un cadre expérimental contrôlé permettant de mettre en évidence et caractériser les capacités d'apprentissage chez la punaise hématophage \textit{Rhodnius prolixus}. \`A la différence des moustiques, les caractéristiques biologiques de cette punaise hématophage, responsable de la transmission de la Maladie de Chagas en Amérique Latine, permettent l'adaptation de protocoles expérimentaux largement validés chez les drosophiles et l'abeille domestique. Nos résultats montrent dans un premier temps que ces insectes sont capables d'apprendre et d'associer la présentation d'une même odeur dite neutre (l'acide lactique), c'est-à-dire qui ne provoque ni attraction ni répulsion lorsqu'elle est présentée seule, avec soit la possibilité d'obtenir une récompense (un repas sanguin, conditionnement appétitif), soit avec la possibilité de recevoir une punition (un choc mécanique, conditionnement aversif). Nous avons également montré que l'apprentissage et la mémoire sont également impliqués dans le choix des hôtes. Les insectes ont en effet associé la présentation d'un choc mécanique avec le complexe d'odeur d'hôtes naturels, biaisant leur préférence lors d'un test de choix réalisé après l'entraînement. Dans un second temps, nous avons adapté à notre modèle d'étude le paradigme de conditionnement de la réponse d'extension du proboscis, développé chez les modèles classiques, ce qui a permis la caractérisation des capacités d'apprentissage, de la durée de rétention à la régulation par des horloges circadiennes. Ces travaux proposent également un paradigme expérimental, reproductible et efficace permettant d'analyser les mécanismes fins qui sous-tendent les processus d'apprentissage et de mémorisation. Dans son ensemble, cette étude apporte la première preuve expérimentale de la capacité d'apprentissage d'insectes vecteurs de la maladie de Chagas et propose des outils expérimentaux et méthodologiques permettant d'améliorer la compréhension des processus associés chez les insectes hématophages en général. Les résultats sont également discutés dans le contexte de la sélection d'hôte et de la transmission des parasites
Learning and memory contribute to animals' fitness by allowing them adapting to variable environments. Thses two processes make them able to extract and use information from their environment in order to reduce the uncertainty associated with unpredictible environments. Among insects, fruit flies and honeybees are considered as classical models for the study of learning and memory. The amount of work that has been done on these models provide a considerable amount of information regarding the genetic, neurobiological and molecular basis of these processes and revealed the complexity of insects' cognitive abilities. All this knowledge acquired in model species, contrasts surprisingly with the lack of knowledge available regarding insect species that are involved in animal and human diseases transmission. Yet, it has been aknowledge that the detailed study of vectors cognitive abilities would allow the understanding of their adaptation to haematophagy, of their vectorial importance and provide new tools for diseases control. Up to date, studies focusing on disaese vectors, mainly in mosquitoes, were conducted in natural or not completely controled contexts and thus no clear demonstration of learning and memory is availaible.The main goal of this work was to provide a controled experimental context allowing the strudy of learning abilities in the haematophagous bug \textit{Rhodnius prolixus}. Our results show that these insectes are able to learn to associate the delivery of a same neutral odour either with the possibility to obtain a reward (blood-meal, appetitive conditioning) or with the possibility to receive a punishment (mechanical shock, aversive conditioning). We also showed that learning ans memory are involved in host selection processes. In a second part, we adapted to our biological model the paradigm of proboscis extension response conditioning, which allowed us to analyse and characterize its learning abilities. The maximal retention duration as well as the modulation of learning abilities by circadian clocks were evinced. Taken as a whole, this work provides the first experimental demonstration of learning abilities in Chagas disease vectors and provides experimental and methodological tools; These latters should allow improving the understanding of the mechanisms that are underlying cearning abilities of haematophagous insects in general. Results are also discussed in the context of host selection and parasite transmission

The cattle tick, Rhipicephalus (Boophilus) microplus is a tick of veterinary and health importance globally, transmitting Babesia bovis and B. bigemina. Tick control is important and needed to prevent livestock diseases caused by tick-transmitted pathogens. Traditionally, tick control methods have resulted in development of acaricide-resistant ticks, environmental pollution and meat and milk contamination. Therefore, there is a need for alternative method and vaccines directed against tick feeding. The aim of this study was to identify proteins involved in tick feeding, tickhost-pathogen interactions and tick reproduction. Consequently, these will help in identification of antigens with the ultimate goal of developing anti-tick vaccines. R. (B.) microplus female ticks were collected at five different feeding stages. RNA was isolated from the salivary gland extracts (SGEs). The cDNA libraries were synthesized and sequenced with the Illumina MiSeq technology. Transcriptome data was analyzed with CLC Genomics Workbench, Trinity and Minia. The SGEs were also used to isolate the fractions: membrane, soluble and pellet protein for proteomic analysis. The proteomics data was analysed with Mascot, X!Tandem and Scaffold. Both the transcriptome and proteome analysis revealed the presence of major secretory protein families such as Kunitz, lipocalins, serpins, cement proteins and metalloproteases, while the majority of transcripts coded for housekeeping genes.
National Research Foundation (South Africa)
Life and Consumer Science
M. Sc. (Life Science)

碩士
台北醫學院
醫學研究所
88
From 1997 to 2000, 160 blood samples(dog 114；cat 2；mouse 12；pigeon 17；goat 3；bat 1；wild boars 3 and flying squirrel 8 ) and 79 stool samples (dog 20；mouse 12；pigeon 30；guinea pig 3；goat 2；wild boars 4 and flying squirrel 8 ) were periodically collected from the animals lived with and hunted by the aboriginal tribes people of 5 villages in Taiwan (Nan Au village in Yi Lan county：Dung Au, Nan Au,Bi Hou,Jin Yue,Wu Ta, Jin Yang and Au Hau；Jin Feng village in Tai Dung county：Jeng Shing,Jia Lan,Shin Shing,Bin Mau and Li Chiou；Hai Duan village in Tai Tung：Hai Duan,Kan Ding,Jia Na Wu Lu,Li Dau,Guang Yuan,Jin Ping andChu Lai；Da Jen village in Tai Tung：Tai Ban,Tu Ban,Shin Hua,An Shuo,Nan Tian, and Sen Yung；Da Wu village in Tai Tung：Shang Wu,Da Wu,Da Ju,Nan Shing, and Da Niau； coast mountain of Hua Lian county). The dog lived with people showed that the infection rates of Toxoplasma gondii, Dirofilaria immitis ,Hepatazoon canis, Ancylostoma spp. ,Toxocara canis and Trichuris vulpis are 21.9%(25/114), 6.1%(7/114), 1.8%(2/114), 35.0% (7/20), 20.0%(4/20), and 5.0%(1/20), respectively. Meanwhile, the Haemoproteus spp. of pigeon is 41.2%(7/17) and the Babesia spp. of mouse is 41.7%(5/12). About the mouse stood, it showed that the infection rates of Ancylostoma spp., Strongyloides ratti and Diphyllobothrium spp. are 16.7%(2/12), 25.0%(3/12) , and25.0%(3/12) , respectively. Meanwhile the multi-infection rates of Strongyloides ratti and Diphyllobothrium spp. are 8.3% (1/12), and 25.0% (3/12). The Strongyloides papillosus of goat is 50.0%(1/2). The Strongyloides ratti of guinea pig is 33.3% (1/3). There were only 2 blood samples of wild boars and flying squirrel. The infection rates of both Toxoplasma gondii and Suifilaris suis of wild boars are 33.3%(1/3). The total parasitic infection rate is 75.0% and 100% in stool samples. The wild boars results showed that all the infection rates are 50.0%(2/4) to Ascaris suum, Trichuris suis, Strongyloides ransomi and Hyostrongylus rubidus, except the Ascarops strongylina is 25.0 %(1/4). The investigation of flying squirrel showed the infection rates of Trichuris spp., Syphacia spp., Coccidia spp., Strongyloides ratti and Nippostrongylus muris are 25.0%(2/8), 87.5%(7/8), 75.0%(6/8), 50.0%(4/8), and 37.8%(3/8), respectively. The 2, 3 and 4 multi- infection of wild boars are 25.0%(1/4). The 2, 3 and 4 multi- infection of flying squirrel are 50.0%(4/8), 25.0%(2/8), and 25.0%(2/8), receptively. In this investigation, the data is accumulated progressively and matched with the current study location and the main purpose is for establishing the animal''s parasitic data of aboriginal tribes'' area in Taiwan and link to Geography Information System. Then the infectious disease, public health, epidemiology and epidemic situation of human and animal in aboriginal tribes could be monitored and controlled instantly and the hygiene education of the tribes could be improved and the parasite disease induced by other disease could also be reduced.

Thesis (M. Agricultural Management (Animal Production)) -- University of Limpopo, 2018
Ethno veterinary medicine practices are popular among the resource constrained small-holder farmers. Nonetheless, the effectiveness of traditional remedies particularly the indigenous plants has not been extensively documented. Hence, this study was conducted to determine efficacy of indigenous plant extracts used by small-holder farmers in Chief Albert Luthuli Local Municipality, Mpumalanga Province, South Africa for internal parasites control. Information on indigenous plants used for the control of internal parasites of livestock by local farmers in the study area was gathered through a questionnaire survey. A total of 188 livestock farmers (both males and females) of mixed ages were interviewed. Thirteen different plants were frequently mentioned by the respondents as remedies for livestock internal parasites. Of these, seven plants could be identified up to their families and species. Plant species collected were Dicerocaryum sp (50%), Pappeacapensis (61%), Aloe ferox (90%), Helichrysum sp (56%), Senecio congestus (83%), Senecio barbertonicus (67%) and Gardenia sp (73%). These plants were extracted using distilled water and analysed to determine their efficacy through in vitro assays; Egg hatch, larval development and larval mortality assays. All the assays were performed at different concentrations of 2.5mg/ml, 5.0mg/ml and 7.5mg/ml. The nematode third stage larvae were incubated for 24hr, 48hr and 72hr during the larval mortality assay. The present study showed that all the seven-plant species under investigation possessed some anthelmintic activities of varying strength. The highest egg hatch inhibition was observed from the extracts of Senecio barbertonicus with 100 % and the lowest from Dicerocaryum eriocarpum with 2.25 %, for larval development the highest was Gardenia sp 100 % and the lowest Helichrysum sp 26 % at concentration 7.5 mg/mL respectively. Whereas, the highest in larval mortality assay was Senecio barbertonicus and Gardenia sp achieved 100 % after 48hrs and the lowest was Dicerocaryum eriocarpum with49.89 % after 72hrs at concentration 7.5 mg/mL respectively. The use of other different forms of extraction media is recommended because different results can observe and be compared with the results of the present study. Toxicity studies on the indigenous plants observed to have stronger anthelmintic activities would assist in the future recommendation of these remedies for large scale or commercial use as anthelmintic drugs. Keywords: ethno veterinary medicine, gastrointestinal parasites, anthelmintic

Pathogens and parasites are increasingly recognized as important components within host populations, communities, and ecosystems. Parasite contributions to ecosystem function most likely manifest as density-mediated impacts of parasites on their hosts, the direct contributions of parasite biomass to a system, and via parasite-induced changes in host behavior and physiology (trait-mediated impacts). Here, a framework was constructed that can be used to conceptualize parasite contributions to ecosystem function (Chapter 1). Then the influence of parasite attack on host movement was explored to further evince the mechanistic underpinnings of trait-mediated parasite impacts (Chapter 2). Additionally, mesocosms were created across a gradient of parasitism to examine how these mechanisms are likely to unfold at larger biological scales (Chapter 3). Lastly, a series of differential equations was created to model host-parasite-ecosystem interactions and generate theoretical predictions about how and when parasites are likely to influence ecosystem processes (Chapter 4). Parasites have many characteristics of ecosystem engineers, but their role has historically been ignored. These studies begin to explore the role that parasitism may have as one of the drivers of ecosystem processes.

Infection of small ruminants by gastrointestinal nematodes (GIN) is a major health concern because they cause substantial economic losses, especially in the tropics and sub-tropics. For many years, control of GIN has been based upon use of anthelmintics. However, there is now a global challange because mutant GIN individuals can tolerate most of the widely used anthelmintics. Therefore, alternative control measures are needed. The objective of the study was to screen a number of plant species for their anthelmintic effects, and to evaluate selected strains of Bacillus thuringiensis (Berliner) and Clonostachys rosea (Schroers) for activity against sheep GIN. Subsequently, the combined treatments would test a dual control strategy for nematodes by using a combination of plant extracts with biocontrol agents. Ethanol extracts of 25 plant species were screened for their anthelmintic effects against Haemonchus contortus (Rudolphi 1803). Extracts of each plant were used in vitro at various concentrations (10, 20 and 30%) to treat 10 day faecal cultures. Five plants with high efficacies (Ananas comosus L. Merr., Aloe ferox Mill., Allium sativum Linn., Lespedeza cuneata Dum. Cours. and Warburgia salutaris Bertol.f. Chiov) were selected for further investigation, using ethanol, dichloromethane and water extracts at four concentrations (2.5, 5, 10 and 20%). Ethanol was the most effective solvent. Larval counts decreased as a result of increasing extract concentrations. An ethanolic extract of Lespedeza cuneata caused more than 70% mortality at all concentrations. In an in vivo study, the five plants A. comosus, A. ferox, A. sativum, L. cuneata and W. salutaris extracts were compared to a positive Control (Equimax®, a modern anthelmintic based on abamectin and praziquantel). Gender, eggs count (EPG₀) and initial body weights were used in assigning sheep (24 females and 24 males) to six groups. Each group was randomly assigned a treatment. Plant extracts were applied as an oral dose (100 mg kg⁻¹ BW), one dose per week per animal for 42 days (Phase 1). Subsequently, the same sheep were dosed for three consecutive days with the same treatments, keeping them in the same groups (Phase 2). Rectal faecal samples were taken for counting of eggs per gram of faeces (EPG) and L₃ larvae per gram (LPG) in faecal cultures. With application of plant extracts, the EPG count decreased with time (P<0.001), and the impact of the plant extracts increased (P<0.001) with time. Two extracts, from A. comosus and L. cuneata, were the most effective in Phase 1 (58% and 61% reduction of EPG, respectively,), and in Phase 2 (77% and 81% reduction of EPG, respectively). In a study on potential biocontrol agents, two strains of Bacillus thuringiensis (Bt) and one of Clonostachys rosea f. rosea (C. rosea), and compared with a diatomaceous earth (DE) product for their anthelmintic activity in sheep. Bacillus thuringiensis and C. rosea were fed to sheep at a rate of 1g kg⁻¹ BW, and DE was fed at 2% of sheep diet. The biocontrol treatments had no effect on EPG (P>0.05), but reduced GIN larvae per gram (LPG) (P<0.001) in faecal culture. Efficacy varied with time (P<0.001). By Day 7 Bt, C. rosea and DE had caused mortalities of GIN of 75.7, 86.9 and 60.6%, respectively. In addition, the efficacy of feeding 1g kg⁻¹ BW of C. rosea chlamydospores to sheep every day, every second day and every third day was tested. Daily feeding of fungal chlamydospores reduced LPG (a count of 12±1.67 GIN larvae) (P<0.001) more than feeding them the biocontrol agent every second day (39±0.77) or third day (58±1.77). By Day 12, feeding the biocontrol agent to sheep every day, every second day, or every third day caused mortality of GIN larvae of 90, 63 and 49%, respectively. Four dietary levels (treatments) of C. rosea (0.25g (F1), 0.5g (F2), 1g (F3) and control (C) of C. rosea product kg-1 BW) were tested. Treatments were each mixed with a complete diet and fed to sheep once daily for 10 weeks, according to body weights. Increased doses of the biocontrol agent reduced LPG (P<0.001), larval development (LD) (P<0.001), and increased efficacy (P<0.001). On Day 70, F1, F2, F3 and the Control controlled LD by 33.3, 72.3, 89.4 and 2.6%, respectively. Clonostachys rosea was effective in reducing third stage larvae (L₃) on pastures significantly (P<0.001) by Day 63 and Day 70. Ethanolic extracts of A. comosus, A. ferox, A. sativum, L. cuneata and W. salutaris all reduced egg production by GIN parasites of sheep. Feeding sheep cultured chlamydospores of a biocontrol fungus, Clonostachys rosea, reduced counts of nematode larvae in sheep; and 1g C. rosea chlamydospores kg⁻¹ BW daily was enough to reduce nematode infective larvae, therefore reducing the degree of pasture contamination. An initial trial showed that the combination of the two treatments of an A. comosus extract and C. rosea chlamydospores was more effective than either treatment on its own in controlling gastrointestinal nematodes in sheep. A long-term trial is being undertaken currently to confirm this finding.
Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.

Text in English
Corridor disease is an economically important tick-borne disease of cattle in southern Africa. The disease is caused by Theileria parva and transmitted by the vectors, Rhipicephalus appendiculatus and R. zambeziensis. There is currently no vaccine to protect cattle against T. parva that is permitted in South Africa. To develop recombinant anti-tick vaccines against Corridor disease, comprehensive databases of genes expressed in the tick’s salivary glands are required. Therefore, in Chapters 2 and 3, mRNA from the salivary glands of R. appendiculatus and R. zambeziensis was sequenced and assembled using next generation sequencing technologies. Respectively, 12 761 and 13 584 non-redundant protein sequences were predicted from the sialotranscriptomes of R. appendiculatus and R. zambeziensis and uploaded to public sequence domains. This greatly expanded the number of sequences available for the two vectors, which will be invaluable resources for the selection of vaccine candidates in future. Further, in Chapter 3, differential gene expression analysis in R. zambeziensis revealed dynamic expression of secretory protein transcripts during feeding, suggestive of stringent transcriptional regulation of these proteins. Knowledge of these intricate expression profiles will further assist vaccine development in future. In Chapter 4, comparative sialotranscriptomic analyses were performed between R. appendiculatus and R. zambeziensis. The ticks have previously shown varying vector competence for T. parva and this chapter presents the search for correlates of this variance. Phylogenetic analyses were performed using these and other publically available tick transcriptomes, which indicated that R. appendiculatus and R. zambeziensis are closely related but distinct species. However, significant expression differences were observed between the two ticks, specifically of genes involved in tick immunity or pathogen transmission, signifying potential bioinformatic signatures of vector competence. Furthermore, nearly four thousand putative long non-coding RNAs (lncRNAs) were predicted in each of the two ticks. A large number of these showed differential expression and suggested a potential transcriptional regulatory function of lncRNA in tick blood feeding. LncRNAs are completely unexplored in ticks. Finally, in Chapter 5, concluding remarks are given on the potential impact the R. appendiculatus and R. zambeziensis sialotranscriptomes may have on future vaccine developments and some future research endeavours are discussed.
Life and Consumer Sciences
Ph. D. (Life Sciences)