Dissertations / Theses on the topic 'Worms – Intestinal and parasitic'

Parasitic platyhelminths are a major burden on humanity through their direct impacts on human health and the huge economic losses they cause, predominantly by undermining agricultural productivity and associated anti-parasite treatments. Previous studies on helminth parasites have highlighted the potential of the neuropeptide signalling system as a source of novel parasite control targets. The use of model organisms is preferable when the target organism is prohibitively difficult or unethical to study directly. The free living microturbellarian Macrostomum lignano is an ideal candidate for the study of parasitic flatworms due to its ease of culture, small size, availability of genomic and transcriptomic datasets and its basal phylogenetic position within the phylum Platyhelminthes. Firstly, this project mapped the distribution of four novel neuropeptides identified from the M. Iignano genome and uses RNA interference (RNAi) on one of those associated neuropeptide genes in an attempt to knockdown the associated transcript and delineate function. Subsequently, the expression of these neuropeptides was examined in selected parasitic flatworms. Despite being successful in the treatment of various parasitic flatworms of animals and humans, the mechanism of action of the anthelmintic praziquantel (PZQ) remains open to debate, hampering efforts to identify mechanisms of resistance and the design of novel drugs which could emulate its effects in parasitic flatworms. The mechanism of action of PZQ was investigated in M. lignano by first determining if the worm was PZQ sensitive. Next the genome was screened for the presence of voltage operated calcium channel (VOCC) β subunit genes (one of the proposed targets of PZQ) which was subsequently silenced using RNA interference to determine if it had a role in PZQ sensitivity in M. lignano. A role for these VOCCs in PZQ action in M. lignano was confirmed using RNAi.

The parasitic nematodes Ancylostoma caninum and Oesophagostomum radiatum were collected and analyzed for intestinal inorganic granules. Three means of identification were utilized to determine the composition granules, including birefringence, x-ray diffraction and energy dispersive spectrometric (EDS) analysis. Initial x-ray diffraction results of the two worms showed a calcium sulfide presence within the worms. Closer examination of the granules within Ancylostoma caninum however, utilizing EDS analysis revealed their composition to be zinc sulfide. These results concur with those of Rogers (1940) and Clark (1956) who found zinc sulfide granules in several species of Strongylus. The ZnS granules seem to be a result of a detoxification function that binds excess zinc and sulfhydryl groups present from the ingestion and breakdown of dietary blood meals.

Thesis (Ph.D.) - University of Glasgow, 2008.
Ph.D. thesis submitted to the Faculty of Biomedical and Life Sciences, Division of Immunology, Infection & Inflammation, University of Glasgow, 2007. Includes bibliographical references. Print version also available.

Intestinal parasitic infections (IPI's) are considered to be a public health problem of global importance by the World Health Organization. The present epidemiologic survey study investigated the prevalence, risk factors, and consequences of pathogenic IPI's on the growth, nutrition and psychomotor development of 244 Ecuadorian children aged 0.2-14 years. The study was conducted in five rural hamlets located in a tropical rainforest area in northwest Ecuador. The study data were obtained by means of a structured questionnaire, a developmental screening examination, anthropometry, and lab analysis of blood and fecal samples. Data analysis was conducted using appropriate bivariate and multivariate statistical techniques. The study results revealed that 90% of the child subjects were infected with at least one pathogenic IPI species. Fifty-one percent were identified with helminthic infections, 37.6% with protozoal infections, and 21.4% were infected with both. The most common intestinal parasites detected were Ascaris lumbricoides (39.7%), Giardia intestinalis (25.2%), Trichuris trichiura (19.7%), Entamoeba histolytica/dispar (18.5%), Blastocystis hominis (13.3%), and Ancylostoma duodenale (1.7%). The prevalence of growth stunting (40%) and iron-deficiency anemia (26%) also was high. Children infected with Giardia exhibited a risk for stunted growth that was twice that of their non-infected counterparts (51.7% vs. 33.1%; OR=2.16, 95% C.I.= 1.13-4.15; p= 0.01). They also had significantly reduced mean blood hemoglobin levels compared to non-infected children 11.8 + 1.5.g/dL vs. 12.2 + 1.4g/dL; p= 0.023) but the proportion with iron-deficiency anemia was slightly but not significantly increased (29.4% vs. 24.3%). The characteristic most consistently associated with risk for pathogenic protozoal IPI's was a high density of domestic animals living in and around the home. Children who lived in such households had a risk for infection that was 2-5 times greater than others. This suggests that domestic animals were important reservoirs for IPI infection in the child group studied. Contrary to the a priori hypothesis, no gender, ethnic, nor age differences in infection risk were identified except for Trichuris infection, which was reduced in younger children contrary to expectations. Mass or targeted chemotherapy combined with health education and promotion are needed to reduce the cycle of infection and re-infection and the negative impact of these on child growth and iron status. Health education and promotion messages can be incorporated into other types of programs already in place in local schools and by the Ecuadorian Ministries of Public Health, Education, and Social Welfare and other agencies.
Master of Science

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Dissertacao (Mestrado)
IPEN/D
Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

Orientador: Regina Maura Bueno Franco
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: As parasitoses intestinais representam um grave problema de Saúde Pública em todo o mundo, com prevalências variáveis em cada região, sendo que fatores socioeconômicos e ambientais corroboram para manutenção ou aumento do número de casos. Os trabalhadores rurais canavieiros, devido as condições as quais são submetidos durante o trabalho sofrem exposições diárias aos agentes parasitários. Eles formam um grupo populacional cuja saúde é negligenciada devido a escassez de informações na literatura. Os objetivos deste trabalho foram: avaliar a prevalência das infecções parasitárias intestinais entre trabalhadores rurais de duas principais regiões canavieiras do Estado de São Paulo (Ribeirão Preto e Campinas) destino de milhares de migrantes oriundos de diferentes regiões do Brasil no período de safra da cana de açúcar; avaliar comparativamente os métodos de coloração de Ziehl Neelsen modificada e Auramina O, para detecção dos oocistos de coccidioses intestinais e, as técnicas de Hoffman, Pons e Janer (H.P.J.) e Coprokit ® para detecção de ovos de helmintos. Foram analisadas 3 amostras fecais de 617 indivíduos, as quais foram processadas pelos métodos de sedimentação espontânea, Faust et al., Rugai et al., Willis et al., Coprokit ®, e pelos métodos de coloração Auramina- O e Ziehl Neelsen modificada, mediante emprego de técnica de centrifugo - concentração a 500X g e, aplicação de um questionário semi- estruturado para avaliar condições sócioeconômicas e epidemiológicas. A prevalência geral de parasitos intestinais e comensais foi de 30,47 %, sendo que o protozoário mais encontrado foi Endolimax nana (12,16 %) e entre os helmintos detectou-se com maior frequência os ancilostomídeos (2,43 %). Dentre os exames positivos, foi verificada maior ocorrência de monoparasitismo (23,82 %) e de protozoários (26,58 %). Conclui-se que a baixa prevalência encontrada para parasitoses intestinais neste estudo se deve as melhorias no perfil socioeconômico e epidemiológico dos trabalhadores canavieiros e entre os exames positivos não houve diferença significativa na detecção de oocistos de Cryptosporidium spp. pelas técnicas de Ziehl Neelsen modificada e Auramina O e, a técnica de sedimentação espontânea se mostrou mais eficiente para detecção de ovos de helmintos em comparação ao método de Coprokit ®
Abstract: Intestinal parasitic diseases represent a serious public health problem worldwide, with prevalence rates varying in each region, and socioeconomic and environmental factors to corroborate maintaining or increasing the number of cases. Rural workers in sugar cane plantations, due to weather conditions and which are submitted during the rural work, and suffer daily exposures to parasitic agents. They form a population group whose health is neglected due to lack of information in the literature. Our objectives were: assess the prevalence of intestinal parasitic infections among rural workers in two major sugar cane growing regions of São Paulo (Ribeirão Preto and Campinas) fate of thousands of migrants from different regions of Brazil from sugar cane crop; benchmarking the methods of Ziehl Neelsen modified and Auramina O for detection of oocysts of intestinal coccidia and the techniques of Hoffman, Pons and Janer (H.P.J.) and Coprokit ® for detection of helminth eggs. Three fecal samples were analyzed from 617 individuals, which were processed by the methods of spontaneous sedimentation, Faust et al., Rugai et al., Willis et al. Coprokit ®, and the staining methods Auramina O and Ziehl Neelsen modified, by use of centrifugal technique - the 500Xg concentration and, application of a semi-structured survey to assess the socioeconomic and epidemiological conditions. The overall prevalence of parasitism was 30.47%, and the most frequent protozoan was Endolimax nana (12,16%) and among the helminthes were detected more frequently the hookworm (2,43%). Among the positive tests, there was verified a higher incidence of monoparasitism (23,82%) and protozoa (26,58%). It is concluded that the low prevalence for intestinal parasites in this study is due to improvements in socio-economic and epidemiological profile of the sugarcane workers and among the positive tests there was no significant difference in detection of oocysts Cryptosporidium spp. by modified Ziehl Neelsen technique and Auramina O, and the spontaneous sedimentation technique was more efficient for detection of helminth eggs in comparison to the method of Coprokit ®
Mestrado
Parasitologia
Mestre em Parasitologia

Foi estudado o perfil nosológico da população ribeirinha do Baixo Rio Machado em Rondônia, área de Amazônia Ocidental no Brasil. Neste estudo, escolhemos abordar o perfil das seguintes doenças: malária, hepatite viral B e C, parasitose intestinal e papilomavírus humano. Estas doenças foram priorizadas devido à importância clínica e epidemiológica e também escassez de estudos referentes nas populações ribeirinhas amazônicas. Para isto, foi realizado um estudo descritivo no Baixo Rio Machado que se localiza a 250 km de Porto Velho (capital do Estado de Rondônia) pela margem direita do Rio Madeira, que é um afluente volumoso do Rio Amazonas, no Brasil. Nesta área, vivem aproximadamente 806 pessoas distribuídas em 55 comunidades ribeirinhas isoladas. Em Rondônia, encontramos locais, como o Rio Machado, que abrigam a presença de portadores assintomáticos do Plasmodium, fato que pode contribuir para a persistência desta doença na região. Para caracterizar o perfil da malária ribeirinha, focalizando aspectos da infecção assintomática e sintomática, foram realizados hemoscopia e Reação em Cadeia da Polimerase (PCR) com amplificação do DNA ribossomal do parasita para Plasmodium vivax e Plasmodium falciparum para o diagnóstico da malária. Foi considerado como assintomático o paciente com hemoscopia positiva ou PCR positivo que permaneceu sem sintomas durante pelo menos 60 dias. Foi realizado acompanhamento longitudinal de 70 dias após o primeiro corte transversal (nC1=585 pessoas) para observação dos pacientes assintomáticos diagnosticados por hemoscopia, onde 25 pacientes (4,25%) permaneceram assintomáticos durante todo esse seguimento. Após, foi realizado mais dois cortes transversais a cada 6 meses (nC2=583 pessoas, nC3=607 pessoas), com tratamento dos indivíduos assintomáticos diagnosticados por PCR no corte transversal anterior ou hemoscopia atual e também dos casos sintomáticos. Foi estudado, na população que permaneceu na área durante toda a pesquisa (n=379), o perfil e o impacto do tratamento da infecção assintomática. Houve diminuição da infecção por P. falciparum de seis vezes e aumento da prevalência de malária por P. vivax de aproximadamente três vezes. O diagnóstico por PCR foi de 2-5 vezes mais eficiente do que por hemoscopia e a prevalência de infecção assintomática foi de 16 a 22% nos cortes transversais. Assim, foi observada a importância do tratamento dos indivíduos assintomáticos para o controle da malária no local, sendo que o incremento de malária por Plasmodium vivax deve ter ocorrido devido a recaídas da doença. Com o estudo entomológico da área, foi verificado que o vetor Anopheles possui atividade hematofágica moderada e principalmente em região peridomiciliar, sendo o A. darlingi a principal espécie da região. Existem poucos dados sobre a prevalência de hepatites na população ribeirinha de Rondônia, o que dificulta a vigilância epidemiológica nestas regiões amazônicas. Foi realizado um estudo de prevalência de hepatite B e C na área, com 123 pacientes pertencentes a 5 comunidades ribeirinhas, incluindo todas as pessoas que estavam presentes na área no momento do corte transversal, de todas as faixas etárias. Foi realizada sorologia para os marcadores de hepatite B: HbsAg, Anti-Hbc (total) e Anti-Hbs e sorologia para hepatite C com o marcador anti-HCV. Foram encontrados 12 (14,7%) pacientes com hepatite B aguda, 29 (38,7%) pacientes apresentaram imunidade vacinal para hepatite B e 7 (9,3%) pacientes apresentaram hepatite C, sendo que um (1,3%) paciente tinha co-infecção para hepatite B e C. Notou-se que esta população ribeirinha está exposta às hepatites virais, sendo necessário intensificar a vigilância epidemiológica na área, assim como a cobertura vacinal e também fornecer cuidados preventivos, curativos e paliativos em relação a estas doenças. A infecção pelo papilomavírus humano (HPV) é altamente prevalente, sendo detectada em aproximadamente 10% a 20% da população sexualmente ativa entre 15 e 49 anos de idade. A introdução de testes mais acurados para a detecção do DNA do HPV em investigações epidemiológicas confirmou a importância do HPV, principalmente dos tipos de alto risco, como o principal fator de risco para o desenvolvimento de neoplasia intra-epitelial cervical e câncer do colo uterino. Não foram encontrados estudos sobre a existência do HPV em populações ribeirinhas amazônicas na literatura médica. Foi realizado, então, um estudo transversal para levantar a prevalência de HPV nesta população, com 84 participantes em idade sexualmente ativa presentes na área. Após coleta de consentimento informado e questionário clínico-epidemiológico, foi realizado exame para isolar o HPV. A tipagem do HPV foi realizada com amplificação dos DNAs por PCR empregando iniciadores genéricos seguida de hibridização em pontos, capaz de identificar mais de 40 tipos diferentes de HPVs. Foram encontradas 18 pacientes contaminadas pelo HPV, perfazendo 21,4% da amostra. Os tipos de HPV encontrados foram: 53, 58, 31, 56, 16, 83, 55, 66, 45, 51, 40, 42, 6, 68. Os tipos de HPV mais freqüentes foram 51 (23%), 58 (19%), 53 (7,7%), 83 (7,7%), sendo HPV 16 encontrado em 3,8% das pacientes HPV positivas e a prevalência do HPV de alto risco oncogênco foi de 13,1%. Desta forma, foi encontrado alta prevalência de HPV na população ribeirinha amazônica estudada, evidenciando a necessidade de vigilância epidemiológica para câncer de colo uterino na região. Por fim, foi estabelecido a prevalência de parasitoses intestinais na população, abordando a correlação das enteroparasitoses com malária assintomática e anemia, realizando um inquérito coproparasitológico na área incluindo pacientes de todas as faixas etárias, que estivessem dispostos a participar do estudo. A análise das amostras foi realizada pelo método da Sedimentação Espontânea (método de Lutz ou Hoffmann, Pons & Janer). Entre os 268 exames de fezes realizados, a prevalência de parasitose intestinal encontrada na região em estudo foi de 86,6%. Entre os helmintos, Ascaris lumbricoides (47%), Ancilostomideos (37,3%), Trichuris trichiura (3,4%), Capillaria hepatica (2,3%), foram os parasitas mais encontrados. Entre os protozoários foram encontrados: Entamoeba coli (21,4%), Entamoeba histolytica (12%), Giardia intestinalis (15,4%), Endolimax nana (10,1%) e Iodamoeba butschlii (7,5%). A alta prevalência de parasitose intestinal encontrada na pesquisa foi concordante com outros dados desenvolvidos na Região Amazônica. Com este estudo, a região da Amazônia Ocidental insere-se no cenário geográfico mundial de distribuição da Capillaria hepatica, particularmente, a região do Baixo Rio Machado.
Was studied the nosological profile of the riverside population of the Baixo Rio Machado in Rondônia, Western Amazon area of Brazil. In this study, was chosed the profile of following diseases: malaria, hepatitis B and C, intestinal parasite and human papillomavirus. These diseases have been prioritized due to clinical and epidemiological importance and scarcity of studies in the riverside Amazon population. For this, was conducted a descriptive study in the Baixo Rio Machado which is located 250 km from Porto Velho (capital of Rondônia State) in the right bank of the Rio Madeira which is a large tributary of the Amazon River in Brazil. In this area, live about 806 people distributed in 55 riversides communities isolated. In Rondônia, we find places such as the Rio Machado, which shelter to the presence of asymptomatic carriers of Plasmodium, which may contribute to the persistence of this disease in the region. To characterize the profile of malaria riverside, focusing on aspects of symptomatic and asymptomatic infections, were performed haemoscopia and Polymerase Chain Reaction (PCR) with amplification of ribosomal DNA for the parasite Plasmodium vivax and Plasmodium falciparum to the diagnosis of malaria. Was considered as asymptomatic patient with positive haemoscopia or positive or PCR that remained without symptoms for at least 60 days. Longitudinal follow-up was conducted for 70 days after the first cross section (nC1 = 585 people) for observation of asymptomatic patients diagnosed by haemoscopia where 25 patients (4.25%) remained asymptomatic throughout the follow-up. After was performed two cross cuts every 6 months (nC2 = 583 people, nC3 = 607 people), with treatment of asymptomatic individuals diagnosed by PCR in previous transverse cross-sections or haemoscopia a current and also symptomatic cases. We studied in the population that remained in the area throughout the study (no= 379), the profile and impact of treatment of asymptomatic infection. There was a decrease in infection by P. falciparum of six times and increased prevalence of malaria by P. vivax approximately three times. The diagnosis by PCR was 2-5 times more efficient than haemoscopia and the prevalence of asymptomatic infection was 16 to 22% in transverse cross-sections. Thus, was saw the importance of treatment of asymptomatic individuals for the control of malaria in the place, and the increase in malaria by Plasmodium vivax should have occurred due to relapse of the disease. With entomological study of the area, was saw that the vector Anopheles has moderate blood activity and especially in peridomiciliary region and the A. darlingi the main species in the region. There are few data on the prevalence of hepatitis in riverside population of Rondônia, which makes surveillance in Amazonian regions. Was conducted a study of prevalence of hepatitis B and C in the area, with 123 patients belonging to 5 communities, including all the people who were present in the area at the cross section of all ages. Serology was performed for markers of hepatitis B: HBsAg, anti-HBc (total) and anti-HBs and serology for hepatitis C with anti-HCV marker. Was found 12 (14.7%) patients with acute hepatitis B, 29(38.7%) patients demonstrated immunity to hepatitis B vaccine and 7(9.3%) patients had hepatitis C, with one (1.3%) patient had co-infection for hepatitis B and C. Realize that this riverside population is exposed to viral hepatitis, being necessary to intensify epidemiological surveillance in the area, as well as immunization coverage and provide preventive care, curative and palliative for these diseases. The infection by human papillomavirus (HPV) is highly prevalent, being detected in approximately 10% to 20% of the sexually active population between 15 and 49 years of age. The introduction of more accurate tests for the detection of HPV DNA in epidemiological investigations confirmed the importance of HPV, particularly types of high risk, as the main risk factor for the development of cervical intraepithelial neoplasia and cervical cancer. Was found no studies on the existence of HPV in riverside Amazonian populations in the medical literature. The transversal cross-section study was take up to raise the prevalence of HPV in this population, with 84 participants in sexually active age in the area. After collection of informed consent and clinical-epidemiological questionnaire, was conducted to isolate the HPV test. The typing of HPV was taking with the DNAs for PCR using generic primers followed by hybridization in points, able to identify over 40 different types of HPV. Was found 18 patients infected by HPV, comprising 21.4% of the sample. The HPV types found were: 53, 58, 31, 56, 16, 83, 55, 66, 45, 51, 40, 42, 6, 68. The most common HPV types were 51 (23%), 58 (19%), 53 (7.7%), 83 (7.7%) with HPV 16 found in 3.8% of HPV positive patients and prevalence the high risk HPV oncogenic was 13.1%. Thus, was find high prevalence of HPV in riverside Amazonian population studied, highlighting the need for surveillance for cancer of the cervix in the region. Finally, was established the prevalence of intestinal parasites in the population, deal the correlation of intestinal with asymptomatic malaria and anemia, taking the fecal examination conducting in the area including patients of all ages, who were willing to participate in the study. The analysis of samples was performed by the spontaneous sedimentation method (method of Lutz or Hoffmann, Pons & Janer). Among the 268 fecal examinations, the prevalence of intestinal parasite found in the region under study was 86.6%. Among helminths, Ascaris lumbricoides (47%), Ancilostomideos (37.3%), Trichuris trichiura (3.4%), Capillaria hepatica (2.3%) were found more parasites. Among the protozoa were: Entamoeba coli (21.4%), Entamoeba histolytica (12%), Giardia intestinalis (15.4%), Endolimax nana (10.1%) and Iodamoeba butschlii (7.5%).The high prevalence of intestinal parasite found in the survey was consistent with other data developed in the Amazon region. In this study, the region of the Western Amazon is part of the scenario of global geographic distribution of Capillaria hepatica, particularly the region of the Baixo Rio Machado.

One of the major constraints limiting the efficiency of sheep production is the control of gastrointestinal nematode parasites. Parasite control by strategic use of anthelmintics is threatened by the emergence of nematode populations that are resistant to the drugs available. It is therefore increasingly apparent that steps toward maintaining sustainable productivity in the growing face of anthelmintic resistance (AR) is required by farmers. To facilitate the uptake of sustainable approaches to parasite management, a comprehensive understanding of the various factors that may influence farmers’ decision-making processes is required. In order to establish which factors are influential, and determine their impact on farmers’ roundworm control behaviours, both qualitative and quantitative research methods were used including focus groups as well as attitudinal and behavioural questionnaires. A retrospective analysis was initially conducted of historical surveys designed to identify farm specific characteristics and parasite management practices including anthelmintic usage. The objectives of the analysis were to identify factors associated with uptake of best practice advice including farm characteristics and information sourcing. In addition, the implementation of sustainable roundworm control practices was investigated using two surveys conducted in 2000 and 2010. Logistic regression models were applied for univariable and multivariable analysis of dependent and independent variables. The next step was to conduct a series of focus groups in different geographic regions of Scotland. The main purposes were to explore sheep farmers’ attitudes towards different aspects of roundworm management, as well as to identify potential motivators and barriers to uptake of sustainable parasite control practices. The findings aided in the development of an attitudinal questionnaire used to canvass opinions representative on a national level. The concluding study involved a telephone survey of 400 Scottish sheep farmers, designed to elicit attitudes regarding roundworm control, AR and sustainable roundworm control practices. A quantitative statistical analysis technique (Structural Equation Modelling) was then used to test the relationships between socio-psychological factors and the uptake of sustainable roundworm control practices. The analysis of historical questionnaire data demonstrated evidence of a shift towards the use of practices to reduce the rate of AR development, most notably a decline in the practice of ‘dose and move’ as well as an overall reduction in treatment frequency. Statistical analysis identified significant associations between farm characteristics and specific treatment strategies. For example, larger farms were more likely to adopt a set treatment regimen (P=0.036), compared with smaller farms, which were more likely to treat based on clinical signs of infection (P=0.021). Sourcing of roundworm control information primarily from veterinarians was most associated with treatment timings with no parallels between time points. From the qualitative focus group studies conducted we identified four overarching themes impacting on sheep producers’ attitudes to roundworm control and best practice advice. These themes comprised: a lack of perceived need to change, the complexity of advice, the ease of implementation of recommended practices and the effectiveness of extension approaches. Additionally, the most important and implementable guidelines identified by sheep farmers were: ‘working out a roundworm control strategy with an advisor’ and ‘administering anthelmintics effectively’ with regard to correct drenching practice i.e. appropriate dosing, administration and drug storage procedures. These findings exhibited similarities with veterinarians’ rankings with also, ‘testing for AR’, ‘preserving susceptible parasites’ and ‘reducing dependence on anthelmintics’ receiving the lowest rankings for importance and implementability. The quantitative analysis from the attitudinal/behavioural questionnaire identified eleven factors with significant influences on the adoption of sustainable roundworm control practices. The key influences on overall adoption were farmers’ baseline understanding about roundworm control and self-reported confirmation of anthelmintic resistance in their flock. Additional positive influences included, positive attitudes to veterinary services, enterprise type and perceived risk of AR. Factors that were shown to have the greatest relative effects on individual parasite control practices included; the perceived resource requirements for implementing a quarantine strategy, farmers’ AR suspicions for instigating AR testing and the confirmation of AR for adopting faecal egg count monitoring. The findings have highlighted several factors which can influence sheep farmers’ decisions to reject or adopt recommended roundworm control practices. It is evident that the perceived complexity, lack of need and practicality of the current recommendations necessitates changes to how extension is designed and disseminated to farmers. The findings also suggest that improving farmers’ acceptance and uptake of diagnostic testing and improving underlying knowledge and awareness about nematode control is a significant target to influencing adoption of best practice behaviours. The importance of veterinarians as highly-trusted information resources validates the need for further engagement with veterinarians concerning sustainable parasite control approaches, to facilitate collaboration with farmers. The need for interaction between farmers and their advisors is key to resolving the issues raised to enable the necessary explanation, justification and execution of recommended practices to suit farmers’ needs and farming conditions.

[ES] La producción en acuicultura se ha visto menguada por aparición de enfermedades en los sistemas de cría de peces. En concreto, en la dorada (Sparus aurata), hay dos parásitos destacados: Enteromyxum leei (Myxozoa) y Enterospora nucleophila (Microsporidia). Hasta la fecha, para ninguno de los dos se ha establecido un cultivo in vitro, y solo para E. leei se ha conseguido establecer un modelo de mantenimiento de la infección in vivo. La presente tesis pretende incrementar el conocimiento sobre estos parásitos y sus relaciones con el hospedador, sentando las bases para generar soluciones que puedan ser aplicadas en la acuicultura. El objetivo con E. leei fue estudiar la inmunidad adquirida inducida en la dorada y la posibilidad de generar herramientas de diagnóstico y vacunas frente a esta enfermedad. Para ello, primero se demostró la resistencia del pez al parásito tras una segunda exposición, la cual duró hasta 16 meses. Además, la resistencia parece estar correlacionada con altos niveles de inmunoglobulina (Ig) M específica en sangre, y una alta expresión de Igs, incluso antes de la re-exposición al parásito. El siguiente paso fue afinar el protocolo de infección con E. leei. Los resultados mostraron que una semana es suficiente para transmitir la infección de E. leei por efluente, independientemente de la temperatura. Tras la demostración de la respuesta adaptativa eficaz frente a E. leei, y al disponer de un modelo de infección refinado, se realizó un ensayo de inmunización pasiva. Aquí, los resultados mostraron que los anticuerpos especi'ficos efectivamente consigue ralentizar la invasión del intestino por el parásito y disminuir los síntomas de la enfermedad. Paralelamente, el resultado del análisis del repertorio de las regiones variables de la IgM e IgT del intestino peces resistentes mostró la inducción de una respuesta policlonal en las ce'lulas B. En base a estos resultados, se realizó una búsqueda de antígenos de E. leei que pudieran ser utilizados como candidatos para la producción de vacunas (análisis proteómico) o herramientas de diagnóstico (análisis in silico). Para ello, se ensambló un transcriptoma de novo utilizando una muestra mixta de intestino de dorada y parásito. Los resultados dieron lugar a 7 y 12 candidatos en la búsqueda in silico y proteómica, respectivamente. En los estudios de E. nucleophila, debido a que fue descrita muy recientemente, el punto de partida fue más básico. Las muestras de este parásito solo se pueden obtener de brotes naturales en piscifactorias. Por ello, primero se realizó un estudio de caracterización de la patología de la infección a partir de peces infectados naturalmente. En etapas tempranas de la infección, el parásito se localiza principalmente en el intestino, pero meses después, la prevalencia en intestino baja e incrementa en los órganos hematopoyéticos y el esto'mago. Los signos clínicos de la infección consistieron en una reducción significativa del crecimiento, emaciación, y palidez de las paredes intestinales. A nivel celular, en los casos ma's graves se observó hipercelularidad en el epitelio intestinal y proliferación de ce'lulas rodlet, un elevado número de linfocitos en la base del epitelio e infiltración de granulocitos acidófilos en el epitelio intestinal. Finalmente se probaron varias formas de transmisión horizontal de E. nucleophila (cohabitación, efluente, intubación oral y anal) con para desarrollar un modelo de mantenimiento in vivo. Se consiguió la transmisión el parásito por todas las vías, pero con una disminución de prevalencia a lo largo del tiempo. Variables como la temperatura, la dosis, y el estado de los peces donantes parecen ser más determinantes que la ruta seleccionada para la transmisión. Entre las rutas probadas, la intubación anal parece ser la más prometedora, pero ninguna de ellas fue capaz de reproducir los signos clínicos observados en las infecciones naturales.
[CA] La producció en aqüicultura s'ha vist minvada per aparició de malalties en els sistemes de cria de peixos. En concret, en l'orada (Sparus aurata), hi ha dos paràsits destacats: Enteromyxum leei (Myxozoa) i Enterospora nucleophila (Microsporidia). Fins avui, per a cap dels dos s'ha establert un cultiu in vitro, i només per a E. leei s'ha aconseguit establir un model de manteniment de la infecció in vivo. La present tesi pretén incrementar el coneixement sobre aquests paràsits i les seves relacions amb l'hoste, establint les bases per a generar solucions que puguin ser aplicades en l'aqüicultura. L'objectiu amb E. leei va ser estudiar la immunitat adquirida induïda en l'orada i la possibilitat de generar eines de diagnòstic i vacunes enfront d'aquesta malaltia. Per a això, primer es va demostrar la resistència del peix al paràsit després d'una segona exposició, la qual va durar fins a 16 mesos. A més, la resistència sembla estar correlacionada amb alts nivells d'immunoglobulina (Ig) M específica en sang, i una alta expressió de Igs, fins i tot abans de la re-exposició al paràsit. El següent pas va ser afinar el protocol d'infecció amb E. leei. Els resultats van mostrar que una setmana és suficient per a transmetre la infecció de E. leei per efluent, independentment de la temperatura. Després de la demostració de la resposta adaptativa eficaç enfront de E. leei, i en disposar d'un model d'infecció refinat, es va realitzar un assaig d'immunització passiva. Aquí, els resultats van mostrar que els anticossos específics efectivament aconsegueix alentir la invasió de l'intestí pel paràsit i disminuir els símptomes de la malaltia. Paral·lelament, el resultat de l'anàlisi del repertori de les regions variables de la IgM i IgT de l'intestí peixos resistents va mostrar la inducció d'una resposta policlonal en les cèl·lules B. Sobre la base d'aquests resultats, es va realitzar una cerca d'antígens de E. leei que poguessin ser utilitzats com a candidats per a la producció de vacunes (anàlisis proteómico) o eines de diagnòstic (anàlisi in silico). Per a això, es va assemblar un transcriptoma de novo utilitzant una mostra mixta d'intestí d'orada i paràsit. Els resultats van donar lloc a 7 i 12 candidats en la cerca in silico i proteòmica, respectivament. En els estudis de E. nucleophila, pel fet que va ser descrita molt recentment, el punt de partida va ser més bàsic. Les mostres d'aquest paràsit només es poden obtenir de brots naturals en piscifactorias. Per això, primer es va realitzar un estudi de caracterització de la patologia de la infecció a partir de peixos infectats naturalment. En etapes primerenques de la infecció, el paràsit es localitza principalment en l'intestí, però mesos després, la prevalença en intestí baixa i incrementa en els òrgans hematopoètics i l'estómac. Els signes clínics de la infecció van consistir en una reducció significativa del creixement, emaciació, i pal·lidesa de les parets intestinals. A nivell cel·lular, en els casos més greus es va observar hipercelularidad en l'epiteli intestinal i proliferació de cèl·lules rodlet, un elevat nombre de limfòcits en la base de l'epiteli i infiltració de granulòcits acidòfils en l'epiteli intestinal. Finalment es van provar diverses formes de transmissió horitzontal de E. nucleophila (cohabitació, efluent, intubació oral i anal) amb per a desenvolupar un model de manteniment in vivo. Es va aconseguir la transmissió el paràsit per totes les vies, però amb una disminució de prevalença al llarg del temps. Variables com la temperatura, la dosi, i l'estat dels peixos donants semblen ser més determinants que la ruta seleccionada per a la transmissió. Entre les rutes provades, la intubació anal sembla ser la més prometedora, però cap d'elles va ser capaç de reproduir els signes clínics observats en les infeccions naturals.
[EN] Aquaculture production is hampered by the emergence of parasite diseases in fish farming systems. Among them, in Sparus aurata, there are two important enteric parasites described: Enteromyxum leei (Myxozoa) Enterospora nucleophila (Microsporidia). To date, no in vitro culture has been established for either parasite, and only for E. leei was it possible to establish a model for maintaining the infection in vivo. The aim of this thesis is to gain new knowledge about these parasites and their relationship with the host, also the basic foundations for generating solutions that can be applied in aquaculture. The general objective for E. leei was to study the acquired immunity induced in gilthead bream and the possibility of generating diagnostic tools and vaccines against this disease. To this end, resistance against the parasite was assessed with a second exposure against the parasite, which showed a resistance for at least 16 months. Besides resistance seemed to be correlated with high levels of specific immunoglobulin (Ig) M in blood, and a high expression of Igs, in particular, the soluble forms, even before re-exposure to the parasite. The next step was refining the protocol for effluent infection with E. leei by studying infection at different exposure time points, temperatures and population densities. The results showed that one week of exposure is sufficient to spread E. leei infection by effluent, regardless of temperature. After demonstrating the resistance against E. leei, and with a refined infection model, a passive immunization assay was performed. The results showed that the serum with specific antibodies effectively slows down the invasion of the gut by the parasite and reduces the symptoms of the disease. At the same time, the analysis of the repertoire of the variable regions of intestinal IgM and IgT showed an induction of a polyclonal response in B cells. On the basis of these results, a research was carried out for E. leei antigens that could have use as candidates for the production of vaccines (proteomic study) or diagnostic tools (in silico study) using the parasite transcriptomic data. To do this, a de novo transcriptome was assembled using a mixed sample of gilthead sea bream and parasite, with a posterior filtrate of the sequences. The In silico and proteomic analysis search resulted in 7 and 12 transcripts, respectively, which are being used for diagnostic and vaccine production. The starting point was more basic in E. nucleophila studies, since this is a recently described disease. The samples of this parasite can only be obtained from natural outbreaks in fish farms. Therefore, first study was carried out to characterize the pathology of the infection of naturally infected fish. In the early stages of the infection, the parasite is mainly located in the intestine, but months later, the prevalence is lower in the intestine and increases in the hematopoietic organs and the stomach. Clinical signs of infection were significant reduction in growth, wasting, and intestinal walls paleness. At the cellular level, in the most severe cases hypercellularity in the intestinal epithelium, proliferation of rodlet cells, high number of lymphocytes at the base of the epithelium and infiltration of acidophilic granulocytes in the intestinal epithelium were observed. Finally, horizontal transmission of E. nucleophila was tried using different transmission methods: cohabitation, effluent, and oral and anal intubation. Transmission of the parasite was achieved with all routes, but there was a decrease in prevalence over time in all cases except for the anal route. Variables such as temperature, dose, and the status of the donor fish appear to be more important than the selected route. Among the routes tested, anal intubation seemed to be the most promising, as it was sustained over a longer period of time, but none of them was able to reproduce the same clinical signs of infection observed in natural infections.
The authors kindly acknowledge the collaboration of anonymous fish farming companies allowing access to the animals during the disease outbreaks. We thank J. Monfort and L. Rodríguez (IATS-CSIC) for the technical assistance on histological processing.This work has been carried out with financial support from the European Union and the Spanish Ministry of Economy and Competitiveness (MINECO) under grant projects ParaFishControl (H2020-634429) and AGL2013-R-48560-C2-2-R, respectively. APS was contracted under ParaFishControl project. Primer sequences and access to the gilthead sea bream transcriptomic database were kindly provided by Prof. J. Pérez-Sánchez of the IATS- Nutrigenomics group. The authors thank I. Vicente for fish maintenance and technical assistance during samplings. The authors thank P. Boudinot (INRAE) for his help in designing and interpreting the immunoglobulin repertoire study and results, J. Pérez-Sánchez (IATS-CSIC) for providing access to the gilthead sea bream genome sequences to perform the repertoire analysis.This work was funded by the European Research Council (ERC Consolidator Grant 2016 725061 TEMUBLYM).
Picard Sánchez, MA. (2021). Control of enteric parasitic diseases of farmed gilthead sea bream: New insights into Enteromyxum leei (Myxozoa) and Enterospora nucleophila (Microsporidia) infections [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/167035
TESIS

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FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais
As parasitoses intestinais são um importante problema de saúde pública principalmente nos países subdesenvolvidos ou em desenvolvimento. Embora o parasitismo intestinal seja amplamente reconhecido como relevante no contexto epidemiológico de diversas comunidades, os estudos sobre o assunto são ainda insuficientes, principalmente no Brasil. Em vista da dificuldade de diagnóstico específico das parasitoses, muitas vezes são realizados tratamentos empíricos com mais de uma droga. O objetivo principal do presente estudo foi avaliar a efetividade e segurança do uso de nitazoxanida no tratamento de poliparasitoses comparado à terapêutica tradicional ofertada pelo serviço público. Além deste objetivo, foi possível investigar a prevalência e os fatores associados às parasitoses intestinais na população de Colônia do Paiol, uma comunidade quilombola, na Zona da Mata, Minas Gerais. Na comunidade analisada, procedeuse um estudo transversal por censo, sendo que dos 425 moradores, 391 (92%) foram avaliados através de questionário estruturado e exame coproparasitológico. Os resultados mostraram uma alta positividade (63,8%), sendo as espécies patogênicas mais freqüentes Ascaris lumbricoides (22,4%) e Trichuris trichiura (17,9%). O poliparasitismo ocorreu em 36,5% dos investigados. O ensaio clínico, controlado, duplo cego, randomizado avaliou 65 indivíduos em dois grupos de tratamento. A taxa de cura foi de 32,4% e 38,7% com a nitazoxanida e com o tratamento convencional, respectivamente, mas esta diferença não foi estatisticamente significativa (p=0,599). A ocorrência de vômito (p= 0,031) esteve associada ao tratamento convencional e de urina esverdeada ao uso de nitazoxanida (p=0,002). Os outros efeitos adversos foram independentes da droga. Houve diferença estatisticamente significativa em relação à cor da pele e a taxa de cura para ambos os tratamentos. A menor eficácia efetividade foi apresentada pelos indivíduos de cor preta. São necessários outros estudos para esclarecer a baixa efetividade nos casos de poliparasitismo, assim como, reavaliar as práticas preventivas e terapêuticas, com o uso de novas drogas e de agentes de largo espectro, podendo a nitazoxanida ser uma droga alternativa neste contexto. Agrega-se às novas possibilidades terapêuticas, a necessidade de políticas públicas que garantam qualidade de vida, através de saneamento básico, educação para saúde e acesso ao sistema público de saúde, minimizando as iniqüidades na sociedade.
Intestinal parasitism is an important public health concern, chiefly in underdeveloped or developing countries. Although widely recognized as a relevant community epidemiological issue, intestinal parasitism has not been sufficiently studied in Brazil. Because specific diagnosis is difficult and generally cumbersome, empiric treatment, sometimes with more than one drug, is frequently employed. The main aim of this study was the assessment of the effectiveness and safety of nitazoxanide for the treatment of intestinal polyparasitism, as compared to traditional therapy provided by the public service. The study also investigated the prevalence of and factors associated with intestinal parasitism in the population of Colônia do Paiol, a quilombola community from the Zona da Mata, Minas Gerais, Brazil. A census-based cross-sectional study used a structured questionnaire and stool examination to assess 391 people (92%) of the 425 inhabitants of the community. The frequency of intestinal parasitism was as high as 63.8%, with predominance of Ascaris lumbricoides (22.4%) and Trichuris trichiura (17.9%). Polyparasitism occurred in 36.5% of those investigated. A double-blind randomized controlled trial assessed 65 individuals in two treatment groups. Cure rates were 32.4% and 38.7% with nitazoxanide and conventional treatment, respectively, but the difference was not statistically significant (p = 0.599). Vomiting (p = 0.031) was associated with conventional treatment and greenish urine with nitazoxanide use (p = 0.002). Other untoward effects were independent of which drug was used. There was a statistically significant difference concerning skin color and cure rates for both treatments. Dark-skinned subjects had lower cure rates. Further studies are necessary to clarify the reasons for the low effectiveness found in these cases of polyparasitism, and to reevaluate preventive and therapeutic approaches with new and broad-spectrum drugs, nitazoxanide being an option in this context. In addition to new therapeutic approaches, there is a clear need to develop public policies that, through the provision of basic sanitation, health education and access to the public health system, assure quality of life and minimize inequity.

La banane plantain (Musa paradisiaca) est l’aliment de base de millions de personnes dans le monde et sa culture génère des revenus permanents pour un grand nombre d’agriculteurs, dans des plantations de taille petite ou moyenne. Comme dans d’autres régions tropicales, la culture du plantain en Guadeloupe et en Haïti est soumise à de fortes contraintes parasitaires aussi bien telluriques (i.e. du sol : nématodes phytoparasites et charançon du bananier) qu’aériennes (Cercosporiose noire notamment). Les moyens de lutte conventionnels reposent sur l’usage de produits de synthèse dont les effets néfastes sur l’environnement (sols, eaux, animaux) comme sur la santé humaine ne sont plus supportables. Il est donc urgent de réfléchir à des solutions agro-écologiques permettant de rétablir les équilibres biologiques, de maintenir une bonne qualité des sols et une production optimale dans les systèmes de culture plantains. C’est le but de ce travail de thèse qui couple la réalisation d’un diagnostic agro-écologique dans des parcelles paysannes, et le test d’alternatives agro-écologiques en milieu semi-contrôlé. Pour ce faire, une typologie des systèmes de culture plantains a été réalisée à l’issue d’une enquête agro-environnementale dans les deux zones d’étude. Elle a permis de sélectionner 23 parcelles en Guadeloupe et 12 en Haïti dans lesquelles un diagnostic agro-écologique a été conduit. Sur la base de ce diagnostic et de la recherche d’alternatives agro-écologiques à l’usage des produits chimiques, une expérimentation au champ a été mise en place en Guadeloupe en station de recherche, permettant le test de trois pratiques culturales innovantes pour le plantain (seules et combinées), à savoir : i) l’introduction d’une plante de service Paspalum notatum pour la gestion des adventices et la réduction de l’utilisation d’herbicides ; ii) l’apport de vermicompost pour le contrôle des nématodes phytoparasites inféodés au bananier plantain et la fertilisation de celui-ci ; iii) l’utilisation de plants sains PIF (Plants Issus de Fragments de tiges) indemnes de nématodes et de larves de charançon du bananier. La typologie des systèmes de culture plantains révèle que : i) en Guadeloupe les précédents : jachère, ananas et banane plantain sont prédominants avec un niveau d’intensification faible (apports d’intrants chimiques faibles et peu fréquents) ou élevé (apports d’intrants chimiques élevés et plus fréquents) ; ii) en Haïti, les précédents : jachère, banane plantain et manioc prédominent avec un niveau d’intensification faible ou nul (apport d’intrants inexistant). Les résultats du diagnostic agro-écologique montrent que, i) lorsque le niveau d’intensification est faible, les bananeraies plantains pérennes et le précédent-ananas permettent de maintenir une bonne qualité du sol et une bonne régulation des parasites telluriques ; ii) lorsque le niveau d’intensification est fort, les populations d’ingénieurs du sol diminuent drastiquement, alors que le cortège parasitaire tellurique augmente sans que cela n’affecte l’obtention de bons niveaux de rendement instantannés (parcelles précédées d’ananas ou d’une jachère principalement) ; iii) en absence totale de fertilisation, il résulte une diminution de l’activité biologique du sol mais aussi du rendement du plantain, exacerbé par le choix des précédents-culturaux (manioc ou banane plantain) en lien avec les contraintes pédoclimatiques et la maladie des raies noires (Cercosporiose noire) causée par Mycosphaerella fijiensis, notamment au sein des parcelles Haïtiennes ; iv) la succession plantain/plantain est la plus pénalisante vis-à-vis de la culture du plantain, car quelque soit le niveau d’intensification, le rendement reste relativement faible, en lien avec une dégradation de l’état sanitaire, comparativement aux autres précédents
Plantain (Musa paradisiaca) is the staple food of millions of people worldwide and its cropping generates ongoing revenues for many farmers who are planting small or medium size areas. As in other tropical regions, plantain cultivation in Guadeloupe and Haiti is under heavy parasitic constraints terrestrial (plant-parasitic nematodes and banana weevil) as well as aerial (black Sigatoka in particular). Conventional means of control based on the use of synthetic products which adverse effects on the environment (soil, water, animals ...) as on human health are not bearable any more. It is thus urgent to think about agroecological solutions allowing to restore the biological balances, to maintain good soil quality and optimal plantain cropping systems.This is the ultimate goal of this thesis which couples the realization of an agroecological diagnosis in peasants’ plots, and the test of agroecological alternatives in semi-controlled conditions. To do this, a typology of plantain cropping systems was carried out from an agrienvironmental survey in the two study areas. It allowed to select 23 plots in Guadeloupe and 12 in Haiti in which an agroecological diagnosis was implemented. Based on this diagnosis and the research of agroecological alternatives to the use of chemicals, a field experiment was set up in Guadeloupe, in an experimental station allowing the test of three innovative practices for plantain cultivation (alone and combined), namely : i) the introduction of a cover-crop Paspalum notatum for weed control while reducing the use of herbicides ; ii) the input of worms’ compost to control plant-parasitic nematodes specific to plantain and to fertilize it ; iii) the use of healthy “PIF” plants (plants issued from stem fragments) free from telluric pests (nematodes and weevil’s larvaes).The typology of plantains cropping systems shows: i) in Guadeloupe the previous crops are: fallow pineapple and plantain predominate with a low level of intensification (low and infrequent chemical inputs) or high (high and frequent chemical inputs); ii) in Haiti, the previous crops are: fallow, plantain and cassava predominate with a low level or no intensification at all (no inputs). The results of the agroecological analysis show that : i) when the level of intensification is low, perennial plantain and pineapple as previous crops help maintaining a good soil quality and a good regulation of the telluric pests ; ii) when the level of intensification is strong, the soil engineers drastically reduce, while the density of telluric parasites increases without affecting good levels of instantaneous yields (plots where the previous crop is pineapple or mostly fallow) ; iii) when the fertilization is totally missing, it decreases the biological activity of the soil furthermore the plantain yields, exacerbated by the choice of the previous crop (cassava or plantain), in connection with soils and climate constraints and the black Sigatoka caused by Mycosphaerella fijiensis, especially in the Haitian plots ; iv) the crop succession plantain/plantain is the most critical regarding the plantain’s cropping, because whatever the level of intensification, the yields remain relatively low in connection with a degradation of the health state, compared to other previous crops.The driving of an experiment in a research station shows that on the scale of one year, the three tested innovative practices allow maintaining a good soils quality. Healthy plants "PIF" have a better health state (absence of plant parasitic nematodes in the roots) which helps a significant increase of the yields. Cover-crop P. notatum helps the weeds and the soil pests control and favors the improvement of soil biological activity and plantain yields. Worms’ compost contributes to the maintenance of a better soils quality while allowing the regulation of the populations of plant-parasitic nematodes of the plantains

This study investigated the prevalences and intensity of intestinal parasites and aspects of their epidemiology among children in the Qwa-Qwa region of the eastern Free St~te. Faecal samples of 1180 children differing socio-economic status from nine schools at altitudes varying from 1660m to 2200m were examined quantitatively by means of the formol-ether sedimentation technique. Socio-economic, and demographic characteristics for the communities served by the schools were obtained from the literature and from a questionnaire. The study showed that, the area supports a markedly low diversity of parasite infections, and at lower intensities, than low altitude areas such as the coastal plain of KwaZulu-Natal and Eastern Cape, the Northern Province, Mpumalanga and the Western Cape. The intestinal parasite fauna affecting children in Qwa-Qwa is dominated by protozoans with only few helminths and no hookworm or bilharzia. The results indicated that factors which influence the transmission of intestinal parasites in Qwa-Qwa appear to be related primarily to social, economic and cultural aspects of the peoples' lifestyles. Climatic factors were not found important. There was a significant seasonal effect on the intensities of all parasite infection, except two protozoans, Entamoeba coli and Endolimax nana. Water source, electricity, house-type and quality of meat were found to be the important socio-economic factors that influenced parasite transmission. These relationships were investigated by fitting logistic regression and generalized linear mixed models. By documenting human parasitism (above 1700m) this study provided an endpoint to the altitudinal transect conducted in 1993 in KwaZulu-Natal by Appleton and Gouws (in press). Public health authorities and Primary Health Care personnel should find this study useful when designing and implementing nutrition and parasite control. Severe ascariasis has been reported from the study area. It will help focus PHC activities in Qwa-Qwa and in the wider context of Free State Province by demonstrating the value of proper personal and environmental hygiene in the home, thereby forming the basis for intestinal parasite control at the community level.
Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1995.

碩士
國立屏東科技大學
野生動物保育研究所
103
Host and parasites coevolve from long-term and dynamic interactions. Infection of parasites is important in shaping the behavioral adaption of host, among which leaf swallowing behavior of carnivore is considered self-medication behavior to defend the infection from intestinal parasites, such as ingestion and defecation of leaves in Chinese lesser civet (Viverricula indica) in Taiwan. This study was aimed to investigate the relationship between the intestinal parasitic infection and leaf swallowing pattern of civets in Jinshueiying National Trail, and 3 captive civets in the Pingtung Rescue Center for Endangered Wild Animals. The occurrence of undigested leaf in feces of the 3 captive civets were recorded for 10 months. Fresh civet feces were collected every month from August 2013 to July 2014 to examine the intensity of parasitic infection and the occurrence of undigested leaves as well as parasite worms. In total, 414 feces sample were inspected, among which 207were collected and analyzed for the parasitic infection, The correlation between parasite infection and monthly mean temperature and rainfall was revealed. 16 species of parasites were detected, including Toxocara sp., Capillaria sp., hookworm, Mesoestoides lineatus, Strongyloides sp. and Trichuris sp. Due to the limitation of identifying parasite species by eggs, 2 group of parasiteeggs still remained unknown. The average infection intensity of hookworms was significantly negatively related to monthly cumulative rainfall, probably caused by landslide effect after rainfall. A significantly positive relationship between average infection intensity of roundworms and average temperature were found, suggesting that suitable temperature may help preserving parasite eggs in feces. Undigested leaves were found in the 3 captive civets’ feces every month, even though no parasite eggs were recovered in feces, suggesting that leaf swallowing behavior is propensity in civets regardless of parasite infection. Furthermore, the occurrence of undigested leaves was significantly associated with the occurrence of the adult roundworm in feces collected in the wild. The results suggest that leaf swallowing by civets is a self-medication behavior, which may help these animals to prevent or control infection by intestinal parasites.

博士
臺北醫學大學
醫學科學研究所
103
Neurophilic parasitic infections (like Toxocariasis and Toxoplasmosis) and intestinal parasites are serious public health problems in many developed and developing countries. Children are particularly at risk due to their attraction to pets (especially dogs and cats), their play habits, poor level of hygiene and in some cases pica. In Nigeria many studies have indicated the existence of Toxocara canis in dogs, but investigations in humans is very scanty and none exist exclusively on children, and in southern Nigeria. With regards to Toxoplasmosis, most of the studies, if not all, in the country are on pregnant women. Hence, there is also paucity of information on the prevalence in children. Intestinal parasitic infections continue to prevail in Nigeria, hence the need for accurate diagnosis. Most studies in the country were done using the Kato-Katz, probably due to cheaper cost, though the technique is believed to be less sensitive compared to the Merthiolate-Iodine-Formaldehyde (MIF) technique. Blood samples were collected (3-5mls) for the detection of both Toxocara canis and Toxoplasma gondii. The larval excretory-secretory antigens, from the third stage larva, were used to test sera for toxocara specific IgG by western blotting. A kit based on latex agglutination (Toxo Test-MT: Eiken Tokyo) was used to test sera for T. gondii and a titre of 1:32 or higher was considered positive. Intestinal helminthes were investigated using stool samples. About 1g of stool sample was put in special containers for the MIF procedure and mixed with 5mls of MIF solution, then passed through a 100 mesh sieve. It was then allowed to stand overnight after which about 50ul was aspirated onto microscope slides and viewed. For the Kato-Katz technique, about 1g of stool was also used. The stool was placed on a filter paper and pressed through a 100 mesh sieve. The sieved stool was then scraped off the sieve surface and placed to fill a hole on a template on a microscope slide. The fecal material was then covered with pre-soaked cellophane strips in malachite green, inverted and pressed for an even spread, then viewed. Out of the 384 samples (blood and stool) examined, results showed that 86.1%, 24% and 86.2% of the children were infected with Toxocara canis, Toxoplasma gondii and intestinal parasites respectively. The MIF technique showed superiority to Kato-Katz in detection of intestinal parasites (p < 0.0001), and it was also able to detect protozoan parasites like Entamoeba hystolytica/E. dispar (27%), Entamoeba coli (10%), Giardia lamblia (13%), Endolimax nana (11%) and Blastocystis hominis (3%), which were not detected by Kata-Katz. With the results of Toxocariasis and Toxoplasmosis infections, we have generated baseline data on these neglected tropical diseases among schoolchildren in Nigeria, while enriching the data on intestinal parasitic infections. There is therefore need for community education on personal hygiene, proper sanitation, and drinking of safe water. Government intervention in these aspects is also urgently required.

The goal of this work was to examine the samples from archeological site in an attempt to identify human and/or animal intestinal parasite eggs. Another task was to detect parasitic protist antigens by ELISA test. Then the results were compared with other facts obtained from research of this locality.

Relatório de Estágio do Mestrado Integrado em Ciências Farmacêuticas apresentado à Faculdade de Farmácia
O presente documento apresenta em primeira instância, no âmbito da unidade curricular “Estágio” do Mestrado Integrado em Ciências Farmacêuticas, os relatórios de estágio em Indústria Farmacêutica e em Farmácia Comunitária. O estágio em Indústria Farmacêutica realizou-se nos Laboratórios Expanscience Portugal, empresa farmacêutica localizada em Lisboa, sendo os Assuntos Regulamentares e Qualidade as áreas em que o meu estágio se concentrou. O estágio em Farmácia Comunitária realizou-se na Farmácia Higiene, localizada no concelho de Torres Novas. Ambos os relatórios apresentam uma análise SWOT (Strenghts, Weaknesses, Opportunities, Threats – Forças, Fraquezas, Oportunidades, Ameaças) dos estágios que realizei nas instituições supracitadas. Ainda como parte integrante deste documento, consta a monografia intitulada “Parasitoses Intestinais: Diagnóstico recorrendo à técnica de Reação de Polimerização em Cadeia em tempo real quantitativa (qPCR) e multiplex qPCR”. O objetivo desta monografia é abordar e comparar a eficácia dos métodos de diagnóstico, aplicados às infeções parasitárias intestinais. Os parasitas intestinais têm sido identificados em todo o mundo. No entanto, é nas regiões tropicais e subtropicais onde a sua prevalência é maior e onde a maioria dos países se encontra em vias de desenvolvimento. As populações destes países apresentam-se, geralmente, infetadas com pelo menos um parasita, não apenas devido à área de elevada endemicidade onde habitam, como também pelas más condições socioeconómicas e maus padrões de higiene. O diagnóstico das parasitoses intestinais nestes países recorre, geralmente, a métodos microscópicos, por serem baratos e simples de executar. Contudo, vários estudos têm-se aplicado em métodos moleculares, como o qPCR e multiplex qPCR, que demonstram ser mais sensíveis e específicos, caracterizando com maior exatidão a epidemiologia das parasitoses intestinais, permitindo a monitorização da eficácia da terapêutica instituída às populações afetadas, tornando possível a implementação de programas de Administração Massiva de Fármacos (MDA) o mais eficientes possível.
This document presents in the first instance, within the scope of the unit "Curricular Internship" of the Integrated Masters in Pharmaceutical Sciences, the internship reports in Pharmaceutical Industry and in Community Pharmacy. The internship in Pharmaceutical Industry was held at Expanscience Laboratories Portugal, a pharmaceutical company located in Lisbon, where Regulatory Affairs and Quality Services were areas in which my internship focused. The internship in Community Pharmacy was held at the Farmácia Higiene, located in the municipality of Torres Novas. Both reports present a SWOT analysis (Strengths, Weaknesses, Opportunities, Threats) of the internships I realized in the aforementioned institutions. Also as an integral part of this document is the Monograph entitled "Intestinal Parasitic Infections: Diagnosis using quantitative real time Polymerase Chain Reaction (qPCR) and multiplex qPCR". The aim of this monograph is to approach and compare the efficacy of diagnostic methods applied to intestinal parasitic infections. Intestinal parasites have been identified all over the world. However, it is in tropical and subtropical regions where its prevalence is higher and where most countries are developing. Populations of these countries are, usually, infected with at least one parasite, not only due to the high endemicity area where they live, but also due to poor socioeconomic conditions and poor hygiene standards. The diagnosis of intestinal parasitic diseases in these countries, usually, involves microscopic methods, since they are inexpensive and simple to perform. However, several studies have been applied on molecular methods such as qPCR and multiplex qPCR, which prove to be more sensitive and specific, characterizing with greater accuracy the epidemiology of intestinal parasitic infections, allowing the monitoring of the efficacy of the established therapy to the affected populations, making possible the implementation of Mass Drug Administration (MDA) programs as efficient as possible.