Academic literature on the topic 'Fishes – Parasitic diseases – Diagnosis'

The Phylum Protozoa brings together several organisms evolutionarily different that may act as ecto or endoparasites of fishes over the world being responsible for diseases, which, in turn, may lead to economical and social impacts in different countries. Apart from the recent advances for the diagnosis of fish diseases in Brazil, little is known on the protozoan parasites and their relationship with environment and host. This revision presents the most important protozoan parasites found in farmed fish from Brazil, not only with emphasis on its diagnosis, biology, transmission and host-parasite relationship, but also on some information that may be useful to researchers in determining the correct diagnosis in fish farms.

SUMMARYAquaculture, including both freshwater and marine production, has on a world scale exhibited one of the highest growth rates within animal protein production during recent decades and is expected to expand further at the same rate within the next 10 years. Control of diseases is one of the most prominent challenges if this production goal is to be reached. Apart from viral, bacterial, fungal and metazoan infections it has been documented that protozoan parasites affect health and welfare and thereby production of fish in marine aquaculture. Representatives within the main protozoan groups such as amoebae, dinoflagellates, kinetoplastid flagellates, diplomonadid flagellates, apicomplexans, microsporidians and ciliates have been shown to cause severe morbidity and mortality among farmed fish. Well studied examples are Neoparamoeba perurans, Amyloodinium ocellatum, Spironucleus salmonicida, Ichthyobodo necator, Cryptobia salmositica, Loma salmonae, Cryptocaryon irritans, Miamiensis avidus and Trichodina jadranica. The present report provides details on the parasites’ biology and impact on productivity and evaluates tools for diagnosis, control and management. Special emphasis is placed on antiprotozoan immune responses in fish and a strategy for development of vaccines is presented.

The global ornamental fish trade is a rapidly growing industry. Cultivation and propagation of ornamental fishes have been increasing in the last 20 years in Iran. Discus (Symphysodon aequifasciatus) from Cichlidae is one of the most popular and expensive aquarium fish. In the past few years farming of this fish has been well developed in Iran. Two breeder discus fish (Symphysodon aequifasciatus) from two different propagation centres (with high mortality) with signs of anorexia, loss of balance, moribundity and darkness in skin colour were referred to Laboratory of Aquatic Diseases of Veterinary Faculty, University of Tehran. After the survey of ectoparasites, necropsy was performed under aseptic conditions; bacterial culture on standard media was done and the alimentary canal was extruded. In both fish no ectoparasite was detected and no bacteria from these cases grew on the standard media. In internal survey 5 and 25 nematodes were detected in each fish. A high number of free eggs were observed in intestine of fish. Regarding morphological characteristics of the nematodes and their eggs, they were identified as Capillaria sp. Treatment of other fish with levamisole was effective and the loss was terminated. Some helminthes like Capillaria pterophylli Heinze, 1933, can cause a high mortality in cichlid aquarium fishes. This study showed that infection with some species of Capillaria could cause a heavy loss in ornamental fish from Cichlidae. Diagnosis of parasites of these fishes can help us to prevent high mortalities.

Background: Piscinoodinium pillulare is a mandatory mastigophore protozoan with no parasitic specificity, and an important ectoparasite of tropical and temperate psiculture. This parasite is responsible for serious health problems in Brazilian native fish. Another important pathogenic protozoan in psciculture belongs to the genus Trichodina spp., which is commonly found on the surface of fish, gills, fins, and integument. The aim of the present study was to describe an outbreak of P. pillulare andTrichodina spp. in three species of fish intensively created, emphasizing the anatomopathological and epidemiological aspects.Cases: Five animals were necropsied, among them three tambaquis (Colossoma macropomu), one pirapitinga (Piaractus brachypomus) and one tilapia (Oreochromis niloticus) from an intensive psciculture in the Federal District. Out of 1500 fishes, 18 (1.2%) became ill and died. It was reported that the animals showed decreased feed intake, discomfort, dyspnea, opercular movements, red skin lesions and consequent death. In the direct examination of scraping of the body surface of a fish, preparedbetween lamina and coverslip, a ciliated ectoparasite, with a circular bell shape, measuring approximately 20-180 μm, adhered, with morphology consistent with Trichodina spp. was evidenced. During necropsy, fragments of different organs were collected and fixed in 10% formalin, routinely processed for histology, embedded in paraffin, cut to five microns thick and stained with hematoxylin and eosin (HE). Macroscopically, there were pale scaly areas in the medial-lateral regions, moderate increase amountof mucus with brown to green lumps on the body surface, partial loss of the fins, swollen gills with whitish mucus and multifocal hemorrhagic areas on the skin and between the scales. Microscopically, moderate multifocal lymphoplasmocytic branchitis was observed with cell hyperplasia and fusion of secondary lamellae associated with trophons (protozoa), consistent with P. pillulare.Discussion: The diagnosis of infection by P. pillulare and Trichodina spp. in this study was based on the epidemiological and clinical-pathological findings. Regarding the first ones, we highlight the variety of fish species affected. P. pillulare is an example of a commensal parasite present on the substrate of culture tanks, and exerts a type of non-obligatory parasitism that uses the fish as a substrate for its fixation under favorable conditions. In this case, only Trichodina spp. was observed during the traditional evaluation of body surface and gills scraping, and histopathology was essential for the verification of the agent P. pillulare. Routine histopathological analysis may also provide a definitive diagnosis, observing the trophons attached to the gill filaments. Atrophy and fusion of secondary lamellae, which promote proliferative lesions in the gills associated with the interlamellar presence of parasites, are responsible for hypoxia, loss of balance and erratic movements. Although they were observed in a small number, Trichodina spp. are believed to be responsible, on a larger scale, for hemorrhagic lesions on the skin and between fish scales. These findings were similar to those of the literature, and the diagnosis of these parasites allows producers and technicians to adopt adequate treatment, as well as prophylactic measures that provide good environmental conditions in order to avoid economic losses.Keywords: fish diseases, protozoa, fish ectoparasites.

Pathological disorders caused due to metazoan parasitic infestation were studied in the hill stream fishes of northwest Himalayan region. Host specificity was found to be one of the fundamental features of metazoan parasites which belonged to the class Cestoda, Nematoda, Trematoda and phylum Acanthocephala. The study indicates that a successful co-evolution of the host and its parasite has caused the adaptation of the later by developing evading mechanisms in order to avoid extinction. Besides this it was also observed that some parasites have even understood to benefit from the well developed antiparasitic armament in fish intestinal epithelia. Thus, parasites are exploiting the antiparasitic response mechanism of the host to optimize, host finding, invasion and survival in the host. Such interaction between host and parasites are considered phylogenetically old. Some monogeneans, cestodes, digeneans and acanthocephalans were found to resist pronounced cellular and host reactions which even improved the attachment of parasite into the host predilection site. Scanning Electron Microscopy and hitstopathological examination was conducted on parasites recovered from fishes in order to understand the host parasite interaction and the damage inflicted by parasite on hill stream fish species.

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Tese (Doutoramento)
IPEN/T
Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

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

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Tese (Doutoramento)
IPEN/T
Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Orientador: Virgínia Bodelão Richini Pereira
Resumo: As infecções intestinais parasitárias são um problema mundial. Parasitas veiculados em água e alimentos como podem ser provenientes da falta de higiene durante o manuseio dos alimentos, contaminação ambiental por material fecal, irrigação de cultivos agrícolas com águas poluídas ou fossas sépticas precárias, situações comuns em países como o Brasil. Tais questões, colaboram para ocorrência de surtos por água e alimentos na população. Assim, o objetivo deste trabalho foi investigar a presença de parasitas importantes em saúde pública em hortaliças e água de irrigação de propriedades do município de Bauru, São Paulo; bem como, nas fezes e nas mãos de manipuladores dos cultivos. As amostras foram coletadas de cinco propriedades do município de Bauru, sendo uma localizada em área urbana e quatro em área rural. Foram obtidas 33 amostras de água de irrigação, 62 de hortaliças, 31 das mãos e dez fecais dos manipuladores. Todas as amostras foram submetidas a análise molecular e as águas de irrigação submetidas ainda a análise microbiológica. Na análise microbiológica foi detectado coliformes totais e E. coli em três propriedades. Na análise molecular, o parasita mais prevalente em água de irrigação e hortaliça foi Cyclospora cayetanensis. Taenia spp. foi detectada em uma hortaliça e Giardia spp. foi mais prevalente nas amostras humanas. Não foi detectado Toxoplasma gondii. As amostras de água de irrigação apresentaram maior quantidade de amostras positivas. Atividades de educação em ... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Parasitic intestinal infections are a worldwide problem. Parasites carried in water and food as they may come from the lack of hygiene during food handling, environmental contamination by fecal material, irrigation of agricultural crops with polluted water or precarious septic tanks, common situations in countries like Brazil. Such issues contribute to the occurrence of outbreaks by water and food in the population. Thus, the objective of this work was to investigate the presence of important parasites in public health in vegetables and irrigation water on rural properties in the municipality of Bauru, São Paulo; as well as in the feces and in the hands of crop handlers. The samples were collected from five rural properties in the municipality of Bauru, one of them located in an urban area and another four in a rural area. Were collected 33 samples of irrigation water, 62 of vegetables, 31 of hands and 10 fecal samples from handlers. All samples were subjected to molecular analysis and irrigation water was also subjected to microbiological analysis. In this microbiological analysis, total coliforms were detected at high rates and E. coli in three of the properties. In molecular analysis, the parasite most prevalent in the analysis and most common in irrigation water and vegetables was Cyclospora cayetanensis. Taenia spp. was detected in one greenery. Giardia spp. was most detected in human samples. Toxoplasma gondii was not detected. The samples of irrigation water had a grea... (Complete abstract click electronic access below)
Mestre

Orientadores: Jancarlo Ferreira Gomes, Alexandre Xavier Falcão
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: As parasitoses intestinais são altamente prevalentes no mundo, estando entre as maiores causadoras de doenças e óbitos em seres humanos. Atualmente, o diagnóstico laboratorial destas parasitoses é realizado por meio de procedimentos técnicos manuais, desenvolvidos na sua grande maioria há décadas, o que justifica a aplicabilidade de técnicas mais sensíveis e práticas para esta finalidade, visando obter resultados eficientes, especialmente em programas governamentais direcionados à Saúde Pública. Sendo assim, o objetivo do projeto foi de avaliar e validar uma nova técnica parasitológica, denominada TF-Test Modified, em comparação com três técnicas parasitológicas convencionais consagradas pela literatura: TF-Test Conventional; Rugai, Mattos e Brisola; e Kato-Katz/Helm-Test. As etapas do trabalho consistiram em realizar coleta de material fecal de 457 indivíduos localizados em regiões endêmicas para parasitoses no município de Campinas, SP; no processamento laboratorial de 1.828 exames; no diagnóstico de 14 espécies parasitárias; e na análise estatística qualitativa de resultados de maneira abrangente. Dentre as espécies parasitárias encontradas, helmintos e protozoários intestinais foram detectados em 42,23% de indivíduos pela técnica de TF-Test Modified, ante 36,76% por TF-Test Conventional, 5,03% por Kato-Katz/Helm-Test, e 4,16% por Rugai, Mattos e Brisola. Destes casos, 54,40% de infecção simples dos indivíduos demonstrou serem portadores de monoparasitismo. A nova técnica parasitológica de TF-Test Modified, quando comparada com as demais técnicas, apresentou alto valor de infecção, como exemplo para dupla, tripla e múltipla, de maneira a perfazer um total de 98,37% de infecções. Ademais, a nova técnica apresentou índice Kappa com grau de concordância Quase Perfeito em todos os parâmetros avaliados com estimativa de 95% (P<0,05), permitiu encontrar com alta eficiência diagnóstica todas as espécies parasitárias estudadas, mostrou um notável diagnóstico verdadeiro, especialmente quando analisada comparativamente com as outras três técnicas convencionais. O atual estudo permitiu concluir que a técnica de TF-Test Modified pode ser utilizada de forma abrangente no diagnóstico qualitativo de protozoários e helmintos intestinais de humanos. O ganho de sensibilidade diagnóstica proporcionada por esta nova técnica deverá ser de estimável contribuição para o diagnóstico individual laboratorial, inquéritos populacionais e controle das parasitoses intestinais, de modo a repercutir em contribuição social
Abstract: Intestinal parasites are highly prevalent worldwide and is among the largest cause of illness and death in humans. Currently, the laboratory diagnosis of these parasites is accomplished through technical procedures manuals, developed mostly for decades, justifying the applicability of more sensitive techniques and practices for this purpose, to obtain effective results, especially in government programs aimed at Public Health. Thus, the objective of the project was to evaluate and validate a new technique parasite, called TF-Test Modified, compared with three conventional parasitological techniques enshrined in literature: TF-Test Conventional; Rugai, Mattos and Brisola, and Kato-Katz / Helm-Test. The steps of the work consisted of conducting a collection of fecal samples from 457 individuals located in regions endemic for parasitic infections in Campinas, SP, in laboratory processing of 1,828 examinations, the diagnosis of 14 parasitic species, and the qualitative statistical analysis of results so comprehensive. Among the species found parasitic, helminths and intestinal protozoa were detected in 42,23% of subjects using the technique of TF-Test Modified, against 36,76% by TF-Test Conventional, 5,03% by Kato-Katz/Helm-Test, and 4,16% Rugai, Mattos and Brisola. Of these cases, 54,40% of single infections of individuals were shown to be carriers of monoparasitism. The new technique parasitological TF-Test Modified compared to other techniques of infection showed a high value, for example double, triple and multiple so as to make a total of 98,37% infections . Moreover, the new technique presented Kappa index level of agreement with Almost Perfect in all parameters with estimated 95% (P <0.05), allowed to meet with high diagnostic efficiency all parasitic species studied showed remarkable true diagnosis, especially when viewed in comparison with other three conventional techniques. The current study showed that the technique TF-Test Modified can be used comprehensively in qualitative diagnosis of intestinal protozoa and helminths of humans. The gain in diagnostic sensitivity afforded by this new technique should be estimable contribution to the individual diagnostic laboratory, population surveys and control of intestinal parasites, in order to reflect on social contribution
Mestrado
Parasitologia
Mestra em Parasitologia

A detecção microscópica de gametócitos pode subestimar sua prevalência e induzir uma avaliação errônea do potencial de transmissão da malária em áreas endêmicas, mantendo desconhecida a proporção de indivíduos infectados que são potenciais transmissores da doença. Este trabalho teve como objetivo a padronização de métodos moleculares para detecção e quantificação de gametócitos de P. falciparum e P. vivax. Descrevemos um método de qRT-PCR, que tem como alvo transcritos do gene pvs25 e pfs25. Detectamos transcritos de pvs25 em 96,4% das amostras sanguíneas provenientes de indivíduos infectados por P. vivax. qRT-PCR foi mais sensível do que a RT-PCR convencional, que tem como alvo o mesmo gene. Descobrimos que a maioria (61,9%) dos portadores de gametócitos são assintomáticos ou tem parasitemias subpatentes e não teriam sido detectados pelas estratégias de controle de rotina da malária. Porém, esses portadores de gametócitos não identificados, geralmente possuem baixas densidades de gametócitos e contribuem com até 4% da carga global de gametócitos na comunidade.
The microscopic detection of gametocytes may underestimate their prevalence, leading to an inaccurate assessment of the potential for malaria transmission in receptive areas and a poor estimate of the proportion of infected individuals who are infectious. We aimed to standardize molecular methods for detection and quantification of gametocytes of Plasmodium falciparum and Plasmodium vivax. Here, we describe a qRT-PCR that targets transcripts of the mature gametocyte-specific pvs25 gene. We found mature gametocytes in 53 of 55 (96.4%) P. vivax infections diagnosed during an ongoing cohort study in a farming settlement located in the southern of Amazonas state. SYBR green qRT-PCR was more sensitive than a conventional RT-PCR that targets the same gene. Most (61.9%) gametocyte carriers were either asymptomatic or had subpatent parasitemias, and would have been missed by routine malaria control strategies. However, potentially undiagnosed gametocyte carriers usually had low-density infections and contributed up to 4% to the overall gametocyte burden in the community.

Larvas de cisticercos de Taenia crassiceps foram deixadas em repouso em TRIS 9mM pH 7,2 com 1mM EDTA até 180 minutos, os sobrenadantes coletados e processados nos 30, 60, 120 e 180 minutos, dando origem aos antígenos de excreção e secreção (ES 30; ES 60; ES 120 e ES 180). A caracterização do antígeno de excreção e secreção de larvas de Taenia crassiceps foi feito por SDS-PAGE e imunoblot utilizando anticorpos monoclonais (AcMos) anti-T. crassiceps e anti-T. solium e anticorpos humanos. Os resultados mostraram que o rendimento do antígeno ES foi menor nos primeiros 30 minutos (0,4 µg) por larva quando comparado os demais ES (ES 60: 2,4 µg; ES 120: 2,9 µg; ES 180: 2,5 µg). O SDS-PAGE confirmou que no ES 30 há menos proteínas. No imunoblot, o ES 180 mostrou que 6 AcMos (anti-LV-Tcra; anti-ES-Tcra; anti-LV-Tso: A3; anti-T-Tso: B4, B11 e A6) reconheceram apenas as frações 18- e 14-kDa do antígeno ES 180. Os AcMos anti-E-Tso (B8) e anti-LV-Tso (B6) não reconheceram frações do antígeno ES 180. Anticorpos presentes em amostras humanas de pacientes com NC reconheceram as frações protéicas entre 94- a 30-kDa e as de 18- e 14-kDa. Utilizando antígeno ES 180 e amostras de soros de pacientes supostamente saudáveis (GC), foram identificadas proteínas acima de 30-kDa e somente uma amostra reconheceu a de 16- kDa, anômala em relação ao perfil 18- e 14-kDa. As amostras de soro de pacientes com outras parasitoses mostraram reatividade com frações ≥ de 30-kDa do ES 180 e o maior índice de reatividade foi com a proteína 71-KDa. Um total de 77%; 70%; 60% e 70% das amostras de pacientes com toxocaríase, esquistossomose mansônica, hidatidose e Chagas, respectivamente, reconheceram a fração 71-kDa do ES 180. O antígeno ES pode contribuir com futuros estudos abordando a complexa relação parasito hospedeiro na cisticercose e na produção de vacinas para o uso em suínos.
Cysticercus Larvae of Taenia crassiceps were maintained in TRIS 9mM pH 7,2 with 1mM EDTA for 180 minutes; the supernatant was collected and processed at 30; 60; 120 and 180 minutes, originating excretion-secretion antigens (ES 30; ES 60; ES 120 and ES 180). The characterization of the ES antigen was conducted through SDS-PAGE and immunoblot using anti-T. crassiceps and anti-T. solium monoclonal antibodies (AcMos) and human antibodies. The results showed that the production of ES antigen was lower in the first 30 min. (0,4 µg) compared with the others (ES 60: 2,4 µg; ES 120: 2,9 µg; ES 180: 2,5 µg). The SDS-PAGE confirmed that ES 30 presented less protein. By immunoblot,6 AcMos (anti-LV-Tcra; anti-ES-Tcra;anti-LV-Tso: A3; anti-T-Tso: B4, B11 and A6) have recognized only the 18- and 14-kDa fractions of the ES 180. The anti-E-Tso (B8) and the anti-LV-Tso (B6) AcMos did not recognize any fractions. Antibodies from human samples NC recognized the proteins from 94- to 30-kDa and from 18- and 14-kDa. Using serum samples of apparently healthy individuals (GC), the ES 180 antigen showed proteins > 30-kDa and one sample recognized the 16-kDa fraction, anomalous when compared to the 18- and 14-kDa fractions. The serum samples of subjects with other parasitoses showed reactivity ≥ 30-kDa, more frequently with 71-KDa protein. A total of 77%; 70%; 60% and 70% of the samples from subjects presenting toxocariasis, esquistossomose mansonic, hydatidosis and Chagas disease, respectively, recognized the 71-kDa fraction of ES 180. The ES antigen may contribute to further studies on the complex cysticercosis parasite/host relation as well as for the production of vaccines for swine.

Orientador: Luiz Jacintho Silva
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas
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Resumo: Os programas de controle da esquistossomose têm obtido relativo sucesso ao controlar a morbidade relacionada a altas cargas parasitárias desta doença, sem, no entanto, diminuir a área de transmissão no Brasil. Como a neuroesquistossomose medular á uma forma grave de esquistossomose não relacionada a altas cargas parasitárias existe risco teórico de ocorrer em áreas de baixa endemicidade. O objetivo deste estudo foi estudar a ocorrência da NE (neuroesquistossomose) medular em uma região de baixa endemicidade, região de Campinas, estado de São Paulo. Foi feito um estudo retrospectivo, descritivo de base hospitalar com busca ativa em múltiplas fontes de informação. Utilizou-se como base os dois maiores hospitais públicos da região de Campinas. Os pacientes com diagnóstico de NE medular tiveram seus diagnósticos ratificados por critérios padronizados e baseados em quadro clínico típico, comprovação da infecção por Schistosoma mansoni e exclusão de outras causas de mielopatia. Os pacientes foram classificados como autóctones, importados, sem informação e indeterminado. Após esta classificação os dados clínicos e epidemiológicos foram analisados. Foram identificados 27 pacientes com NE medular dos quais 19 (85,2%) homens e 4 (14,8%) mulheres, as idades no momento do diagnóstico foram de 13 a 57 anos (média=31,2; desvio padrão=12,8 e mediana=29). Os pacientes foram classificados quanto ao local provável de infecção da seguinte forma: 14(51,9%) autóctones, 11(40,7%) importados e 2(7,4%) sem informações, não houve paciente classificado como indeterminado. Todos os pacientes importados se infectaram em municípios de áreas de alta endemicidade. A clínica deste grupo de pacientes não foi diferente do encontrado na literatura, nem foi diferente quando comparados os pacientes autóctones com os importados. Houve uma demora média de 70,6 dias (mediana=19; dp=166,9) entre a primeira consulta e o diagnóstico. A demora foi em média 88,1 dias maior entre os pacientes autóctones (média=112 dias; mediana=26; dp=224,3) do que entre os importados (média=23,9 dias; mediana=9; dp=42,7) e esta diferença foi estatisticamente significativa p=0,0247. A sensibilidade da sorologia foi de 87,5%, da imunologia de LCR 93,8% e dos exames parasitológicos foi de 40,0%. Apenas 4 (14,8%) tiveram evolução com melhora completa, 6 (22,2%) apresentaram melhora sem limitações, 13 (48,1%) apresentaram melhora com limitações e 4 (14,8%) não apresentaram melhora alguma. Apenas 11 pacientes (41%) com NE medular incluídos neste estudo estavam notificados à vigilância epidemiológica e a informação de que estes pacientes tinham quadros neurológicos não constavam no banco de informações do SINAN. Concluiu-se que a NE medular ocorre mesmo em áreas de baixa endemicidade e nestas áreas existe uma demora muito grande no diagnóstico, principalmente entre os pacientes autóctones. O exame de fezes não se mostrou sensível para diagnóstico e rastreamento de pacientes vulneráveis a NE medular por se tratarem de pacientes com baixas cargas parasitárias. Portanto métodos diagnósticos mais sensíveis deveriam ser utilizados pelos programas de controle de esquistossomose
Abstract: Programs for schistosomiasis control have enjoyed relative success in controlling death associated to high parasitary loads for this illness, without, however, decreasing the area of transmission in Brazil. Since spinal neuroschistosomiasis is a grave form of neuroschistosomiasis unrelated to high parasitary loads, there is a theoretical risk of its occurrence even when not in a particularly endemic area. The goal of this study was to study the occurrence of spinal NE (neuroschistosomiasis) in a non-endemic area, the region of Campinas, in the Sate of São Paulo. A retrospective, descriptive, hospital-based study was carried, with information actively sought after from various sources of information. The two largest public hospitals in the region of Campinas were used as bases. The patients diagnosed with spinal NE had their diagnoses ratified according to standard criteria and based on typical clinical status, proof of infection by Schistosoma mansoni and the exclusion of other causes for myelopathy. Patients were classified as autochthonous, imported, without information and undetermined. After this classification, the clinical and epidemiological data were analyzed. A total of 27 patients with spinal NE were identified, of which 19 (85.2%) were men and 4 (14.8%) women. The ages on diagnosis ranged from 13 to 57 (average=31.2; standard deviation=12.8 and median=29). The patients were classified as to their probable location of infection the following way: 14(51.9%) autochthonous, 11(40.7%) imported and 2(7.4%) without information. No patients were deemed undetermined. All imported patients were infected in municipalities located in highly endemic areas. Clinical evaluation of this group of patients was no different from that found in the literature, nor was it different when autochthonous patients were compared to imported patients. There was an average period of 70.6 days (median=19; sd=166.9) between the first consultation and diagnosis. The period was on average 88.1 days longer for autochthonous patients (average=112 days; median=26; sd=224.3) than for imported patients (average=23.9 days; median=9; sd=42.7) and this difference was statistically significant p=0.0247. Sensitivity of the serology was 87.5%, LCR immunology 93.8% and for parasitological exams it was 40.0%. Only 4 (14.8%) had evolution with complete recovery, 6 (22.2%) presented improvement without limitations, 13 (48.1%) presented improvement with limitations and 4 (14.8%) did not present improvement. Only 11 patients (41%) with spinal NE included in the study had been notified to epidemiological surveillance and the information that these patients had neurological patterns of symptoms was not present in the SINAN data base. It can be concluded that spinal NE occurs even in non-endemic areas and that diagnosis in such locations can take excessively long, especially for autochthonous patients. Feces exams were not shown to be sensitive for diagnosis and tracing of patients vulnerable to spinal NE since such patients presented low parasitary loads. Therefore more sensitive means of diagnosis should be utilized by schistosomiasis control programs
Mestrado
Ciencias Biomedicas
Mestre em Clinica Medica

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.

Introducción: Las infecciones por parásitos transmitidos por vía fecal-oral son frecuentes en áreas remotas de la Amazonia. No obstante, la epidemiología de estos parásitos en la Amazonia y la validez de técnicas de laboratorio tradicionales empleadas en el diagnóstico permanecen hasta hoy poco estudiadas. Objetivos: Estudio 1: Estudiar la dinámica de la transmisión de Calodium hepaticum en una comunidad rural del municipio de Presidente Figueiredo, estado del Amazonas, Brasil; Estudio 2: Estimar la prevalencia de parásitos intestinales y estudiar factores de riesgo para las infecciones por helmintos en una cuidad remota (Barcelos) del municipio de Barcelos, estado del Amazonas; Estudio 3: Evaluar el desempeño de dos técnicas de sedimentación espontánea (Técnica de sedimentación espontánea de Lutz -SST- y Paratest) para el diagnóstico de parásitos intestinales. Material y métodos: Para el estudio 1 se analizaron muestras de heces de 135 personas (y 2 perros) y tejido hepático de un pecarí. Las heces de los perros fueron recogidas del jardín de los domicilios de personas que presentaron infección espuria por C. hepaticum. Fueron investigados los hábitos alimentarios de la población. Para el estudio 2, un estudio transversal fue realizado con 594 personas de 137 domicilios. Fueron recogidos datos socio-demográficos y características de los domicilios. En el estudio 3, una única muestra de heces de 143 individuos fue evaluada realizando tres réplicas para cada prueba. Fueron evaluados los resultados de la homogeneidad y concordancia entre réplicas y entre pruebas. La sensibilidad y especificidad fueron estimadas usando un modelo de clase latente bayesiano. Resultados: Estudio 1: La incidencia poblacional de infección espuria fue del 6.7% (IC 95%: 2.08 - 11.24) y la intradomiciliar entre el 50 a 83.3%. El riesgo de infección espuria fue 10 veces mayor en personas que consumen hígado de mamífero silvestre (P = 0.02). El hígado del pecarí y una muestra de heces de perro presentaron huevos de C. hepaticum. El consumo del hígado infectado fue identificado como la causa de las infecciones espurias diagnosticadas en uno de los domicilios. Estudio 2: Las prevalencias de Ascaris lumbricoides, anquilostoma, complejo Entamoeba histolytica, Trichuris trichiura y multiparasitosis por helmintos fueron del 53.4%, 27.1%, 13.6%, 8.8% y 24.6%, respectivamente. Los principales factores de riesgo detectados fueron no tener letrina (OR= 1.71, IC 95% 1.05 - 2.78) para la infección por A. lumbricoides; ser varón (OR= 2.43, IC 95% 1.58 - 3.75) y tener suelo de tierra o madera en el domicilio (OR= 1.74, IC 95% 1.04 - 2.90) para la infección por anquilostoma; finalmente ser varón para la infección múltiple por helmintos (OR= 2.26, IC 95% 1.30 - 3.93). Fueron identificados cuatro barrios con mayor riesgo de infección por helmintos. Estudio 3: Ambas pruebas presentaron una alta repetibilidad para la detección de parásitos intestinales exceptuando para Blastocystis spp. y anquilostoma. La concordancia entre pruebas fue alta (coeficiente de correlación de concordancia, 0.72 - 0.88) excepto para Blastocystis spp., anquilostoma y T. trichiura. Las dos pruebas presentaron una baja sensibilidad (39.4% - 49.2% para SST, 35.8% - 53.8% para Paratest) y una alta especificidad (93.2% - 97.2%). Conclusiones: Hemos descrito una alta incidencia intradomiciliar de C. hepaticum relacionada al hábito de comer hígado de mamífero silvestre y este mecanismo de transmisión fue importante en la dinámica de transmisión del helminto en la comunidad. La identificación de factores de riesgo y barrios de mayor riesgo de infección por helmintos en la ciudad puede ayudar en la implementación de medidas específicas para el control de las infecciones. Las dos técnicas de sedimentación presentaron elevada repetibilidad y concordancia en general, baja sensibilidad y elevada especificidad para el diagnóstico de parásitos intestinales.
Introduction: There are few studies on the epidemiology of parasites transmitted via the fecal-oral route in remotes areas of Amazonia and the validity of the laboratory techniques used to diagnose these parasites. Objectives: (1) Study the dynamics of the transmission of Calodium hepaticum in a community of the state of Amazonas, Brazil. (2) Estimate the prevalence of intestinal parasites and study the risk factors of helminth infections in a city of the state of Amazonas. (3) Evaluate the performance of the Lutz spontaneous sedimentation technique and the Paratest for diagnosis. Material and methods: (1) Stool samples of 135 people (and 2 dogs), the liver of a peccary and the alimentary habits of the population were evaluated; (2) Stool samples from 584 subjects were evaluated and sociodemographic data collected; (3) Stool samples (n=143) were evaluated performing three replicates/test. Homogeneity results and concordance between replicates and tests were assessed. Sensitivity and specificity were estimated using a Bayesian latent class model. Results: (1) The incidence of spurious infection was 6.7% (95% CI: 2.08-11.24) and the intradomicile incidence was 50%-83.3%. The risk of infection was 10-fold greater in persons consuming wild mammal liver (P=0.02). The consumption of peccary liver caused infection in one household. (2) The prevalence of Ascaris lumbricoides, hookworm and Trichuris trichiura was 53.4%, 27.1% and 8.8%, respectively. The main risk factors were lack of latrine (OR=1.71; 95% CI 1.05-2.78) for A. lumbricoides infection; being male (OR=2.43; 95% CI 1.58-3.75) and having a earth or wood floor at home (OR=1.74; 95% CI 1.04-2.90) for hookworm; being male for multiple infection by helminths (OR=2.26; 95% CI 1.30-3.93). Four neighborhoods presented greater risk of helminth infection. (3) Both tests presented a high repeatability except for Blastocystis spp. and hookworm diagnosis. Concordance between tests was high except for Blastocystis spp., hookworm and T. trichiura. Both tests presented a low sensitivity (35.8%-53.8%) and a high specificity (93.2%-97.2%). Conclusions: We found a high intradomicile incidence of C. hepaticum related to the habit of eating mammal liver. Identification of risk factors may help to implement control measures. Both tests showed little sensitivity for the diagnosis of intestinal parasites.