Eukaryotic expression and antigen epitope prediction of the LRRC15 protein in excretory secretory antigens of Taenia solium cysticercus / 中国血吸虫病防治杂志

OBJECTIVE@#To conduct eukaryotic expression of the leucine-rich repeat containing 15 (LRRC15), a differentially expressed protein in excretory secretory antigens of Taenia solium cysticercus, and predict its antigen epitope.@*METHODS@#The molecular weight, stability, amino acid sequence composition, isoelectric point and T lymphocyte epitope of the LRRC15 protein were predicted using the bioinformatics online softwares ExPASy-PortParam and Protean. The full-length splicing primers were designed using PCR-based accurate synthesis, and the LRRC15 gene was synthesized. The recombinant pcDNA3.4-LRRC15 plasmid was constructed and transfected into HEK293 cells to express the LRRC15 protein. In addition, the LRRC15 protein was characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.@*RESULTS@#The recombinant pcDNA3.4-LRRC15 plasmid was successfully constructed, which expressed the target LRRC15 protein with an approximately molecular weight of 70 kDa. Bioinformatics prediction with the ExPASy-PortParam software showed that LRRC15 was a hydrophilic protein, which was consisted of 644 amino acids and had a molecular weight of 69.89 kDa and an isoelectric point of 5.6. The molecular formula of the LRRC15 protein was C3073H4942N846O953S28 and had an instability coefficient is 50.3, indicating that LRRC15 was an instable protein. Bioinformatics prediction with the Protean software showed that the dominant T-cell antigen epitopes were located in 292 to 295, 353 to 361, 521 to 526 and 555 to 564 amino acids of the LRRC15 protein, and the T-cell antigen epitopes with a high hydrophilicity, good flexibility, high surface accessibility and high antigenicity index were found in 122 to 131, 216 to 233, 249 to 254, 333 to 343, 358 to 361, 368 to 372, 384 to 386, 407 to 412, 445 to 450, 469 to 481, 553 to 564, 588 to 594, 607 to 617 and 624 to 639 amino acids. Following transfection of the recombinant pcDNA3.4-LRRC15 plasmid into HEK293 cells, SDS-PAGE and Western blotting identified LRRC15 proteins in cell secretory culture media, cell lysis supernatants and sediments. The LRRC15-His fusion protein was purified from the cell culture medium, and SDS-PAGE identified a remarkable band at approximately 70 kDa, while Western blotting successfully recognized the band of the recombinant LRRC15 protein.@*CONCLUSIONS@#The eukaryotic expression and antigen epitope prediction of the LRRC15 protein in the excretory secretory antigens of T. solium cysticercus have been successfully performed, which provides insights into further understandings of its biological functions.

Neurocisticercosis: lesión única cerebral con serología negativa y confirmación post exéresis. Reporte de caso. / Neurocysticercosis: single brain lesion with negative serology and post exeresis confirmation. Case report

Los diagnósticos diferenciales de una lesión única intraparenquimatosa cerebral con borde en anillo son múltiples, desde un proceso infeccioso parasitario hasta una neoplasia primaria del sistema nervioso, por lo que su manejo puede ser controversial y debe tenerse en cuenta la prevalencia epidemiológica según la situación demográfica donde se presente. De esta manera, presentamos el caso de una paciente adulta joven que ingresó al servicio de neurocirugía con sintomatología focal sensitiva asociado a cefalea intermitente, con una TC de encéfalo que reveló una lesión subcortical localizada en la parte superior del giro postcentral izquierdo con edema perilesional importante y realce periférico tras contraste. Se realizaron pruebas de serología en suero y LCR para neuroparásitos, las cuales se reportaron negativas. Se procedió a realizar exéresis completa de la lesión con uso de ecografía intraoperatoria para minimizar tiempo quirúrgico y área de lesión cortical, la cual se envió a estudio de histopatología y reportó el diagnóstico de neurocisticercosis en estadio nodular granulomatoso. Se realiza el presente reporte de caso con una búsqueda exhaustiva en la literatura sobre la sensibilidad y utilidad de las pruebas de serología parasitaria para la confirmación diagnóstica de la neurocisticercosis y para evitar intervenciones neuroquirúrgicas invasivas

The differential diagnoses of single, ring-enhancing brain lesions are multiple, from a parasitic infection to a primary tumor of the central nervous system, therefore, the management of these lesions can be controversial, and the epidemiological prevalence must be taken into account according the demographic situation of the patient. We present the case of a young adult female who was admitted to the neurosurgery service, debuting sensory focal symptoms associated with headache. Brain CT and MRI revealed a subcortical cyst-like lesion in the left superior postcentral gyrus, with large perilesional edema and peripheral enhancement following contrast administration. Serological test for CNS parasites in serum and CSF were negative. Consequently, we realized a complete removal of the cyst with intraoperative ultrasound to optimize the surgical time and the cortical surgical area. The histopathological examination revealed a degenerating cysticercus. For this reason, a bibliographic research was carried out to identify the sensitivity of serological tests for the diagnostic confirmation of neurocysticercosis and thus avoid invasive neurosurgical interventions

Molecular phylogenetic analysis of Cysticercus ovis from Egypt based on MT-CO1 gene sequences / Análise filogenética molecular de Cysticercus ovis do Egito com base nas sequências do gene MT-CO1

Abstract Cysticercus ovis or sheep measles is the larval stage of Taenia ovis, which is the intestinal tapeworm of dogs. It is found in the cardiac and skeletal muscles of sheep and can be the cause of partial or total condemnation of carcasses at abattoirs. The aim of the current work was to determine the prevalence of C. ovis among sheep in Upper Egypt and to present the molecular and phylogenetic analysis of this using the amplified Mitochondrial Cytochrome Oxidase subunit 1 (MT-CO1) gene. A total of 1885 sheep slaughtered at local abattoirs of 4 different governorates of Upper Egypt (Asuit, Sohag, Qena and Aswan) were carefully examined for C. ovis. The overall prevalence of infection was 2.02%. The highest rate of infection was observed in adult animals over 4 years of age (44.73%). There was no significant effect of animal sex on infection rates. The phylogenic analysis of C. ovis Egyptian isolates showed very close similarity to the New Zealand isolate (AB731675). This is the first report showing the genetic analysis of C. ovis in Egypt, which provides a very powerful tool for taxonomy and definitive diagnosis of C. ovis, which could be helpful for preventive and control programs.

Resumo Cysticercus ovis "sheep measles" é o estágio larval da Taenia ovis, encontrada nos músculos de carneiros, causado pela ingestão de ovos de Taenia ovis, parasita de cães. O objetivo do presente trabalho foi determinar a prevalência de C. ovis entre ovinos no Alto Egito e apresentar as análises moleculares e filogenéticas, utilizando o gene da subunidade mitocondrial citocromo-oxidase amplificada 1 (MT-CO1). Um total de 1885 ovinos abatidos em matadouros locais de 4 províncias diferentes do Alto Egito (Asuit, Sohag, Qena e Aswan) foram cuidadosamente examinados para C. ovis. A prevalência geral de infecção foi de 2,02%. A maior taxa de infecção foi observada em animais adultos com mais de 4 anos de idade (44,73%). Não houve efeito significativo do sexo nas taxas de infecção. A análise filogenética de isolados egípcios de C. ovis mostrou uma similaridade muito próxima ao isolado da Nova Zelândia (AB731675). Este é o primeiro relato mostrando a análise genética de C. ovis no Egito, fornecendo uma ferramenta para taxonomia e diagnóstico definitivo de C. ovis, podendo ser útil para programas preventivo e de controle.

Caracterização das lesões parasitárias de ovinos observadas na linha de abate / Characterization of parasitic lesions of sheep observed at slaughter line

Considerando a possibilidade de erros na identificação das lesões e a necessidade de melhorar o diagnóstico, o objetivo desse trabalho foi caracterizar macroscopicamente e microscopicamente as principais lesões parasitárias observadas em ovinos na linha de abate. Os materiais foram colhidos durante duas visitas a um matadouro frigorífico de ovinos no estado do Rio Grande do Sul. Totalizaram-se 161 amostras com lesões parasitárias de ovinos em diferentes órgãos. As alterações observadas incluíam hidatidose, cisticercose por Cysticercus ovis, cisticercose por Cysticercus tenuicollis, sarcocistose (morfologia compatível com Sarcocystis gigantea), fasciolose (Fasciola hepatica) e esofagostomose. Das 161 amostras, 25,5% correspondiam a hidatidose, e os cistos hidáticos foram observados, predominantemente, nos pulmões (46,3%) e fígado (41,5%). Ao corte, os cistos demonstraram três padrões morfológicos: cistos uniloculares viáveis (34%); cistos multivesiculares viáveis (31,7%); e cistos hidáticos (uniloculares e multivesiculares) degenerados (34%). As lesões de cisticercose por C. ovis (22,4%) foram visualizadas no coração (63,9%), língua (13,9%), músculo masseter (11,1%) e diafragma (11,1%). Morfologicamente os cisticercos foram classificados em vivos (viáveis), degenerados e mineralizados. Lesões provocadas por S. gigantea (19,2%) estavam presentes na túnica muscular do esôfago, na língua e na laringe. Macroscopicamente, observaram-se múltiplas estruturas nodulares brancas que continham uma cápsula fibrosa e lúmen preenchido por material gelatinoso translúcido. Cisticercose por C. tenuicollis representaram 18,6% das lesões. Os cistos foram observados aderidos no omento, no mesentério, na cápsula do fígado e na vesícula biliar. Morfologicamente os cistos foram classificados como vivos (viáveis) e degenerados. Os cistos vivos apresentavam a parede translúcida ou levemente opaca, contendo um escólex no seu interior. Os cistos degenerados eram brancacentos, firmes, com espessa cápsula fibrosa e centro mineralizado. Lesões provocadas por F. hepatica corresponderam a 7,4% dos casos. As lesões hepáticas caracterizavam-se macroscopicamente por espessamento variável dos ductos biliares por fibrose e ocasionalmente havia exemplares de F. hepatica no lúmen dos ductos. Em oito casos, observaram-se áreas de acentuada necrose do parênquima hepático. Lesões provocadas por Oesophagostomum spp. perfizeram 6,8% dos casos. As alterações foram observadas no intestino delgado e intestino grosso de todos os ovinos e em dois casos, havia também envolvimento dos linfonodos mesentéricos. Nos intestinos, as lesões caracterizavam-se por nódulos bem delimitados, salientes na serosa, firmes e que invadiam também a camada muscular. Nos linfonodos havia obliteração do parênquima nodal por acentuada mineralização. É extremamente importante a identificação morfológica das diferentes lesões parasitárias encontradas nas linhas de abate em frigoríficos de ovinos, para posterior destino correto das mesmas. As alterações devem ser avaliadas com o intuito principal de reconhecer a sua capacidade infecciosa. Além disso, é fundamental o conhecimento dos locais anatômicos mais comuns em que cada alteração geralmente costuma ocorrer.(AU)

Considering the possibilities of mistaken diagnoses in identifying lesions at meat inspection this study was designed to provide data for a better-educated diagnosis by the meat inspectors through the gross and microscopic characterization of parasitic lesions observed in slaughtered sheep at the inspection line. One hundred and sixty-one samples of parasitic lesions were sampled from various organs of slaughtered sheep during two visits to a sheep abattoir located in the state of Rio Grande do Sul, Brazil. Lesions observed included hydatid cysts, cysticercosis due to Cysticercus ovis and to Cysticercus tenuicollis, sarcocystosis (morphology compatible with Sarcocystis gigantea), fasciolosis (Fasciola hepatica) and oesophagostomosis. Twenty-five point five percent of the 161 samples corresponded to hydatidosis and the hydatid cysts were observed predominantly in the lungs (46.3%) and liver (41.5%). On cut surface, the cysts had three different morphological patterns: viable unilocular cysts (34%); viable multivesicular cysts (31.7%); and degenerate (unilocular and multivesicular) hydatid cysts (34%). Cysticercosis by C. ovis (22.4%) was observed in the myocardium (63.9%), tongue (13.9%), masseter (11.1%), and diaphragm (11.1%). Morphologically the cysticerci were classified as viable, degenerated or mineralized. Lesions caused by S. Gigantea (19.2%) were observed in the muscle layer of the esophagus, tongue, and larynx. Grossly there were multiple white nodular structures that contained a fibrous capsule with the lumen filled by translucent and gelatinous material. Cysticercosis by C. tenuicollis accounted for 18.6% of observed parasitic lesions; the cysts adhered to the omentum, mesentery, liver capsule, and serosal surface of gall bladder; grossly the cysts were classified as viable and degenerated. Viable cysts had translucent or slightly opaque walls and contained a single scolex. Degenerated cysts were white, firm and with a thick fibrous capsule and mineralized center. Lesions caused by F. hepatica accounted for 7.4% of the cases and were grossly characterized by variable fibrous thickening of bile ducts which occasionally contained the adult flukes in their lumina. In eight cases there were marked areas of necrosis in the hepatic parenchyma. Lesions caused Oesophagostomumspp. accounted for 6.8% of the observed parasitic cases and the changes were observed in all cases in the walls of the small and large intestine; in two cases mesenteric lymph nodes were also involved. In the intestines, lesions were characterized by firm well-circumscribed nodules prominent in the serosal surface and also invading the muscle layer. In the lymph nodes marked mineralization obliterated the nodal parenchyma. The correct identification of the various parasitic lesions found in the viscera of sheep in the abattoir inspection line it is important to dictate the proper destination of affected organs and carcasses. The lesions should be evaluated aiming to determine their infective capacity and to acquire knowledge about their more frequent anatomical sites.(AU)

Impact of the neglected Cysticercus cellulose infection on mental health among school-aged children in Tibetan agricultural areas / 中南大学学报(医学版)

To determine the impact of Cysticercus cellulose (C. cellulose) infection on mental health among school-aged children in Tibetan agricultural areas of Sichuan Province.
 Methods: In October 2015, all primary schools located in Tibetan agricultural areas in Yajiang, Ruoergai, and Muli county of Sichuan Province were selected as the research sites. All school-aged children at five- and six-grade were enrolled for the study by a multistage stratified cluster sampling method. Antibodies against C. cellulose were detected. Mental Health Test and questionnaire survey were conducted for school-aged children to collect data. The impact of C. cellulose infection on mental health among school-aged children was analyzed with the multilevel linear regression.
 Results: A total of 2 453 school-aged children were investigated. The C. cellulose seropositive rate was 6.03% (148/2 453). There were 0.16% (4/2 453) patients with seropositive accompanied by seizure, 2.28% (56/2 453) with seropositive accompanied by headache, 2.08% (51/2 453) with seropositive accompanied by frequent weak, and 0.41% (10/2 453) were seropositive accompanied by frequent nausea. The rate of C. cellulose infection was 4.53% (111/2 453). The mean score of the mental health test was 6.59±2.61. There were significant difference in score of mental health test in children whose demographic characteristics were different. The mental health scores of school-aged children were clustered at the school level. After controlling the factors of demographic characteristics, the result of multilevel model demonstrated that the factor of school-aged children with C. cellulose seropositive accompanied by headache was statistically significant (β=1.14, P=0.017).
 Conclusion: The status of C. cellulose infection among school-aged children in Tibetan agricultural areas is not optimistic. C. cellulose infection has impacted on mental health of local school-aged children. It is necessary to strengthen the prevention and control of C. cellulose infection in epidemic area.

Aspectos macroscópicos e histológicos da cisticercose bovina / Macroscopic and histological findings of bovine cysticercosis

Cisticercose bovina é uma importante doença parasitária de caráter zoonótico, com elevada ocorrência em algumas regiões do Brasil. Considerando a possibilidade de erro na identificação das lesões, bem como a dificuldade de classificação dos cistos e a necessidade de melhorar o diagnóstico, o objetivo desse trabalho foi caracterizar e correlacionar as lesões macroscópicas e microscópicas da cisticercose bovina, além de utilizar a técnica da PCR para auxiliar na identificação do agente. Amostras de lesões císticas e nodulares de bovinos, compatíveis macroscopicamente com cisticercose, foram coletadas em abatedouros frigoríficos do Estado do Rio Grande do Sul. Os cistos foram divididos em três grupos: Grupo 1, cisticercos vivos (viáveis); Grupo 2 (subdividido em 2a e 2b), cisticercos degenerados com potencial escólex viável; e Grupo 3 cisticercos mortos (mineralizados). Após a obtenção das lâminas histológicas dos cisticercos de cada grupo, foi realizada a correlação macroscópica e microscópica. Para a realização da técnica da PCR foram utilizadas lesões císticas de 26 bovinos. Foram analisados cisticercos de 127 bovinos, totalizando 232 cistos. Dos 127, 46 bovinos (36,2%) apresentaram mais de um cisticerco e 81 (63,8%) um cisticerco cada. Em relação a localização anatômica dos cistos, o coração demonstrou o maior envolvimento (55,9%), seguido do músculo masseter (22,8%). Quando houve o envolvimento de dois órgãos em um mesmo bovino, coração e músculo masseter juntos, totalizaram 11 casos (8,6%). De maneira geral a média da frequência de cisticercose foi de 10% a 15% de bovinos acometidos por lote. Entretanto, a média isolada de alguns lotes demonstrou condenações acima de 50%, 80% e 90%. Morfologicamente, os 232 cisticercos foram classificados dentro de três grupos. No Grupo 1, 23 cistos (9,9%) foram considerados como vivos (viáveis), e eram caracterizados por lesões císticas com parede translúcida ou levemente opaca, contendo líquido claro e um ponto esbranquiçado no interior (escólex). Na histologia, os cistos eram compostos por uma membrana de onde invaginava um escólex de Taenia saginata. No segundo grupo (Grupo 2), foram incluídos 156 (67,2%) cisticercos degenerados com potencial escólex viável e macroscopicamente os cistos demonstraram dois padrões morfológicos distintos. No primeiro deles (Grupo 2a), visualizado em 111 casos (71,1%), observaram-se lesões nodulares com aspecto caseoso. Microscopicamente, os cistos caracterizavam-se por formações nodulares compostas por área central contendo escólex e membrana, ambos degenerados, e necrose caseosa. No segundo padrão (Grupo 2b), observado em 45 cisticercos (28,9%), as lesões também eram caseosas, entretanto ao corte os cistos demonstravam na área central um orifício em meio ao material caseoso. Os aspectos microscópicos dos 45 cistos incluídos no segundo padrão macroscópico assemelhavam-se aos cisticercos do primeiro padrão. Entretanto, oito cistos (17,8%) demonstraram somente a membrana parasitária viável e em um cisto notou-se a membrana com o escólex viável. No restante dos 36 cistos (80%), observou-se área central contendo escólex e membrana, ambos degenerados, e necrose caseosa. No terceiro grupo de classificação morfológica dos cisticercos (Grupo 3), foram inseridos os cistos mineralizados (mortos), totalizando 53 cistos (22,9%). O aspecto macroscópico desses cisticercos caracterizava-se por lesões nodulares, amarelas, firmes ao corte, que se fragmentavam. Histologicamente observaram-se formações nodulares com área central de acentuada mineralização, rodeadas por infiltrado inflamatório granulomatoso. Dos 127 bovinos, foi realizado PCR a partir do DNA extraído dos cisticercos de 26 bovinos, no qual 24 foram positivos para cisticercose. Em relação aos dois cisticercos negativos, um deles fazia parte do Grupo 2a e o outro do Grupo 3. A correlação entre os aspectos macroscópicos e microscópicos do segundo padrão morfológico observado dentro do Grupo 2, demonstrou que esse subgrupo representa o maior problema na interpretação, pois alguns cistos apresentaram características de viabilidade. Macroscopicamente esses cisticercos podem ser identificados quando cortados, porque possuem um orifício na área central que pode auxiliar no diagnóstico.(AU)

Bovine cysticercosis is an important zoonotic parasitic disease with high prevalence in several regions of Brazil. Considering the need of improvement of the accuracy of diagnosis of these lesions, as well as the difficulty of classification of the cysts, this study aimed to correlate gross and histopathological changes of bovine cysticercosis and to use polymerase chain reaction (PCR) as an aid in their identification. Cystic and nodular lesions from cattle, grossly compatible with cysticercosis, were sampled in slaughterhouses from Rio Grande do Sul State. Lesions were allotted in one of the following groups. Group 1: viable cysticercus; Group 2 (subdivided 2a e 2b): degenerating cysticercus with a potentially viable scolex; and Group 3: dead cysticercus (mineralized). The gross and microscopic aspects of every cysticerci of each group were compared. Two hundred and thirty two cysts and nodules compatible with cysticercus were sampled from 127 bovine. Twenty six of those lesions were tested with PCR. Out of 127 cattle, 46 (36.2%) had more than one cyst and the remaining 81 (63.8%) had on cyst each. Myocardium was the most frequently involved anatomical site (55.9%), followed by masseter muscle (22.8%). When there was more than one organ involved in the same bovine, myocardium a master muscle sum up 11 cases (8.6%). In general, the average of cysticercosis frequency was 10-15%. However the average in some cattle lots was in excess of 50%, 80% and 90%. Morphologically, 232 cysticerci were classified in three groups. In Group 1, 23 cysticerci (9.9%) were considered viable and were characterized by cysts of translucent or slightly opaque wall, containing clear and a white point (scolex) within the cyst. Histologically, the cysts consisted of a membrane from which a scolex of Taenia saginata invaginated. One hundred and fifty six cysts (67.2%) were allotted in Group 2; grossly these cysts revealed two different morphological patterns. In 111 (71.1%) cases of Group 2 (Group 2a) nodular caseous lesions were observed. Histologically, the cysts were characterized by nodules consisting by a central area containing the scolex and membrane, both degenerated, and caseous necrosis. In the remaining 45 (28.9%) cases of Group 2 (Group 2b), lesions were also caseous; however, at cut surface the cysts had a central hole amidst the caseous material. The microscopic aspect of the 45 cysts included in the second was similar to that of the first pattern. However in eight (17.8%) of the 45 cysts only a viable parasitic membrane was observed and in one cyst the membrane and viable scolex were observed. In the remaining 36 cases (80%), the cysts consisted of a central area containing both degenerated membrane and scolex, and caseous necrosis. In Group 3, 53 dead cysts (mineralized) (22.9%) were found among the total of 232 cysts. The gross aspect of these cysticerci was characterized by yellow form nodules which crumbled when cut. Histologically nodules were observed with marked central area of mineralization surrounded by granulomatous inflammatory response. Twenty four of the twenty cysts examined by PCR were positive for Cysticercus bovis and two of them were negative. One of the negatives was part of Group 2 (degenerated cysts) and the other one of the Group 3 (dead mineralized cysts). The correlation between gross and microscopic aspects of the second morphologic aspect of the Group 2 demonstrated that this subset represents a major complicating factor in interpretation, since a large number of these cysts reveal characteristics of viability. Grossly, these cysticerci might be identified when cut, since a hole in the central area will be observed aiding in recognizing his lesions.(AU)

Prevalence and risk factors for Cysticercus tenuicollis in goats and sheep in Paraíba, northeastern Brazil / Prevalência e fatores de risco para Cysticercus tenuicollis em caprinos e ovinos na Paraíba, Nordeste do Brasil

Abstract This study aimed to determine the prevalence and risk factors for C. tenuicollis among goats and sheep in slaughterhouses in Paraíba. 390 animals (195 goats and 195 sheep) in the municipalities of Patos and Esperança, Paraíba, Brazil, were inspected between February and May 2014. The prevalence of C. tenuicollis was 39% (76/195) in goats and 17.4% (34/195) in sheep. In both species, most of the cysticerci vesicles were located at the omentum and mesentery. The only risk factor found was extensive sheep farming. It can be concluded that C. tenuicollis is highly prevalent in small ruminants in Paraíba, being more prevalent in goats than in sheep. Extensively-reared sheep were twice as likely to develop infection by this parasite.

Resumo Objetivou-se determinar a prevalência e os fatores de riscos de C. tenuicollis em caprinos e ovinos nos matadouros da Paraíba. Foram inspecionados 390 animais (195 caprinos e 195 ovinos) nos municípios de Patos e Esperança, Paraíba, Brasil, no período de Fevereiro a Maio de 2014. A prevalência de C. tenuicollis em caprinos foi de 39% (76/195) e em ovinos de 17,4% (34/195). Os locais mais acometidos por C. tenuicollis nas duas espécies foram o omento e o mesentério. O único fator de risco encontrado foi a produção extensiva de ovinos. Pode-se concluir que é alta a prevalência de C. tenuicollis em pequenos ruminantes no Estado da Paraíba, sendo mais prevalente na espécie caprina; e que ovinos criados de forma extensiva apresentam duas vezes mais riscos de contrais a infecção por este parasita.

Experimental neurocysticercosis: absence of IL-4 induces lower encephalitis / Neurocisticercose experimental: ausência de IL-4 induz menos encefalite

ABSTRACT Neurocysticercosis (NCC) is the most severe clinical manifestation of cysticercosis. One of the factors responsible for its symptomatology is the host inflammatory response. Therefore the influence of interleukin 4 (IL-4) on the induction of encephalitis in experimental NCC was evaluated. Methods BALB/c (WT) and BALB/c (IL-4-KO) mice were inoculated intracranially with Taenia crassiceps cysticerci and euthanized at 7, 30, 60 and 90 days later, the encephala removed and histopathologically analyzed. Results The absence of IL-4 induced greater parasitism. In the initial phase of the infection, IL-4-KO showed a lower intensity in the inflammatory infiltration of polimorphonuclear cells in the host-parasite interface and intra-parenquimatous edema. The IL-4-KO animals, in the late phase of the infection, showed lower intensity of ventriculomegaly, encephalitis, and meningitis, and greater survival of the parasites in comparison with the WT animals. Conclusion The absence of IL-4 induced lower inflammatory infiltration, ventriculomegaly and perivasculitis in experimental NCC.

RESUMO A Neurocisticercose (NCC) é a manifestação clínica mais severa da cisticercose, e um dos fatores responsáveis pela sintomatologia é a resposta inflamatória do hospedeiro. Desta forma avaliou-se a influência da interleucina 4 (IL-4) na indução de encefalite na NCC experimental. Métodos Camundongos das linhagens BALB/c (WT) e BALB/c (IL-4-KO) foram inoculados intracranialmente com cisticercos de Taenia crassiceps e eutanasiados aos 7, 30, 60 e 90 dias após a infecção, os encéfalos foram removidos e analisados histopatologicamente. Resultados A ausência da IL-4 induziu um maior parasitismo nos animais. Na fase inicial da infecção os animais IL-4-KO apresentaram menor intensidade tanto de infiltrado inflamatório de polimorfonucleares na interface parasito-hospedeiro quanto de edema intraparenquimatoso. Os animais IL-4-KO, na fase tardia, apresentaram menor intensidade de ventriculomegalia, encefalite, meningite e maior sobrevivência dos cisticercos em relação aos animais WT. Conclusão A ausência da IL-4 induz menos infiltrado inflamatório, ventriculomegalia e perivasculite na NCC experimental.

Inferior rectus muscle ocular cysticercosis: a case report

Cysticercosis is a systemic parasitic disease caused by the larval form of cestode Taenia solium. It has a worldwide distribution and is potentially harmful with variable clinical manifestations. The most commonly involved sites include eye, brain, bladder wall, and heart. Ocular cysticercosis can be extraocular or intraocular and may present with varied clinical symptoms. We report the condition in a thirteen year old female child who presented with mild lower lid swelling and diplopia in upgaze, wherein cysticercus cellulosae cyst was found within the mass of the right inferior rectus muscle. It becomes important to report this case because of the relative rarity of the condition these days, unusual site of the cyst and the young age of the patient

Sequence Analysis and Molecular Characterization of Wnt4 Gene in Metacestodes of Taenia solium

Wnt proteins are a family of secreted glycoproteins that are evolutionarily conserved and considered to be involved in extensive developmental processes in metazoan organisms. The characterization of wnt genes may improve understanding the parasite's development. In the present study, a wnt4 gene encoding 491amino acids was amplified from cDNA of metacestodes of Taenia solium using reverse transcription PCR (RT-PCR). Bioinformatics tools were used for sequence analysis. The conserved domain of the wnt gene family was predicted. The expression profile of Wnt4 was investigated using real-time PCR. Wnt4 expression was found to be dramatically increased in scolex evaginated cysticerci when compared to invaginated cysticerci. In situ hybridization showed that wnt4 gene was distributed in the posterior end of the worm along the primary body axis in evaginated cysticerci. These findings indicated that wnt4 may take part in the process of cysticerci evagination and play a role in scolex/bladder development of cysticerci of T. solium.

Identificación morfológica y molecular de Cysticercus fasciolaris aislado de un roedor (Rattus norvegicus) de la provincia de Buenos Aires (Argentina) / [Morphological and molecular identification of Cysticercus fasciolaris isolated from rodent hosts (Rattus norvegicus) in Buenos Aires province (Argentina)].

In a rodent (Rattus norvegicus) survey in Buenos Aires province, metacestodes of tapeworms were found encysted in the liver of the host. The aim of this work was the morphological and molecular identification of this parasite. To achieve the molecular characterization of the parasite, ribosomal (28S) and mitochondrial (COI) DNA were amplified and sequenced. Based on both morphological and molecular data using bioinformatic tools, the metacestode was identified as Cysticercus fasciolaris. The adult form of this tapeworm (Taenia taeniaeformis) commonly infects felid and canid mammalian hosts. This is the first report on the molecular identification of Cysticercus fasciolaris in Buenos Aires province (Argentina).

Comunicación de un caso de cisticercosis subcutánea / Report of case of subcutaneous cysticercosis

Infection of the larval form (cysticerco) of Taenia in any tissue or organ is known as the disease cysticercosis. Many sites of infection have been documented but the central nervous system has been the most common. It present a case report of a 19 years old patient with a subcutaneous cysticercosis confirmed with biopsy.

La infección por la forma larvaria (cisticerco) de Taenia solium en cualquier tejido u órgano se conoce como cisticercosis. Existen numerosos reportes de casos, siendo la mayoría de ellos cisticercos en sistema nervioso central. El compromiso de otros órganos es raramente detectado. Se presenta el caso de una mujer de 19 años con una cisticercosis subcutánea que fue confirmada con biopsia.

Epidemiological survey of the taeniasis/cysticercosis complex in cattle farms in Viçosa County, Minas Gerais, Brazil / Inquérito epidemiológico sobre o complexo teníase-cisticercose bovina no município de Viçosa, Minas Gerais

Despite the importance of human taeniasis/bovine cysticercosis for public and animal health and the economy, its actual epidemiological status in Brazil is not well-studied. We sought to assess the epidemiological profile of this zoonosis in the rural area of Viçosa County, Minas Gerais, Brazil. The prevalence of bovine cysticercosis was 0.42%, whereas no case of human taeniasis was diagnosed. Factors favoring the persistence of zoonosis were identified. These included the supply of untreated water to animals, animals raised for slaughter without sanitary supervision, poor mechanization of raising techniques, and the use of untreated water for human consumption. Bovine meat for human consumption acquired in the city or from the farm itself was characterized as a risk factor for bovine cysticercosis (Odds Ratio (OR) =16.77; p<0.05). Nevertheless, the families on the investigated farms did apply several appropriate measures to control this disease, such as the virtual lack of open sewers and the consumption of well-cooked meat. The presence of bovine cysticercosis cases, and the factors favoring its persistence, point to the need for constant epidemiological and sanitary surveillance in this county.

Apesar da importância do complexo teníase-cisticercose bovina para a saúde pública, animal e para a economia, a realidade epidemiológica da ocorrência dessas zoonoses no Brasil é pouco conhecida. Este estudo foi realizado com o objetivo de avaliar o perfil epidemiológico desta zoonose na zona rural do município de Viçosa, MG. A prevalência encontrada de cisticercose bovina foi de 0,42%, e nenhum caso de teníase humana foi diagnosticado. Foi verificada presença de fatores favoráveis a manutenção dessa zoonose, como fornecimento de água sem tratar aos animais, criação de animais destinados ao abate sem inspeção sanitária, criação de bovinos com baixa adoção de tecnologia, e utilização de água sem tratamento. O consumo de carne bovina oriunda da propriedade e da cidade foi caracterizado como variável de risco (OR=16,77; p<0,05) para a cisticercose bovina. Apesar disso, as famílias das propriedades pesquisadas possuíam hábitos favoráveis como medida de controle para o desenvolvimento desta parasitose, como a quase ausência de esgoto a céu aberto e a ingestão de carne bovina bem passada. A presença de caso de cisticercose bovina e de fatores considerados favoráveis a manutenção dessa parasitose, mostra a necessidade da contínua vigilância epidemiológica e sanitária neste município.

Evaluation of cysticercus-specific IgG (total and subclasses) and IgE antibody responses in cerebrospinal fluid samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies / Avaliação das respostas de anticorpos anti-cisticercos IgG (total e subclasses) e IgE em amostras de líquido cefalorraquidiano de pacientes com neurocisticercose apresentando produção intratecal de anticorpos específicos IgG

In the present study, an enzyme-linked immunosorbent assay (ELISA) standardized with vesicular fluid of Taenia solium cysticerci was used to screen for IgG (total and subclasses) and IgE antibodies in cerebrospinal fluid (CSF) samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies and patients with other neurological disorders. The following results were obtained: IgG-ELISA: 100% sensitivity (median of the ELISA absorbances (MEA)=1.17) and 100% specificity; IgG1-ELISA: 72.7% sensitivity (MEA=0.49) and 100% specificity; IgG2-ELISA: 81.8% sensitivity (MEA=0.46) and 100% specificity; IgG3-ELISA: 63.6% sensitivity (MEA=0.12) and 100% specificity; IgG4-ELISA: 90.9% sensitivity (MEA=0.85) and 100% specificity; IgE-ELISA 93.8% sensitivity (MEA=0.60) and 100% specificity. There were no significant differences between the sensitivities and specificities in the detection of IgG-ELISA and IgE-ELISA, although in CSF samples from patients with neurocysticercosis the MEA of the IgG-ELISA was significantly higher than that of the IgE-ELISA. The sensitivity and MEA values of the IgG4-ELISA were higher than the corresponding values for the other IgG subclasses. Future studies should address the contribution of IgG4 and IgE antibodies to the physiopathology of neurocysticercosis.

No presente estudo, uma reação imunoenzimática (ELISA) padronizada com o fluido vesicular de cisticercos de Taenia solium foi utilizada para avaliar as respostas de anticorpos anti-cisticercos IgG (total e subclasses) e IgE em amostras de líquido cefalorraquidiano (LCR) de pacientes com neurocisticercose apresentando produção intratecal de anticorpos específicos IgG e pacientes com outras desordens neurológicas. Os seguintes resultados foram obtidos: ELISA-IgG: 100% de sensibilidade (mediana das absorbâncias das reações ELISA (MAE)=1,17) e especificidade 100%; ELISA-IgG1: sensibilidade 72,7% (MAE=0,49) e especificidade 100%; ELISA-IgG2: sensibilidade 81,8% (MAE=0,46) e especificidade 100%; ELISA-IgG3: sensibilidade 63,6% (MAE=0,12) e especificidade 100%; ELISA-IgG4: sensibilidade 90,9% (MAE=0,85) e especificidade 100%; ELISA-IgE: sensibilidade 93,8% (MAE=0,60) e especificidade 100%. Não foram encontradas diferenças significativas entre as sensibilidades e especificidades das reações ELISA-IgG e ELISA-IgE, embora a MAE da reação ELISA-IgG em amostras de LCR de pacientes com neurocisticercose tenha sido significativamente maior que a obtida com ELISA-IgE. Os valores de sensibilidade e MAE da reação ELISA-IgG4 foram maiores que os valores correspondentes para as outras subclasses da IgG. Estudos futuros deverão abordar a contribuição dos anticorpos IgG4 e IgE na fisiopatologia da neurocisticercose.

Human Neurocysticercosis Case and an Endemic Focus of Taenia solium in Lao PDR

A male patient with neurocysticercosis was identified in Montai Village, Xay District, Oudomxay Province, Lao PDR in February 2004. He had a history of diagnosis for neurocysticercosis by a CT scan in Thailand after an onset of epileptic seizure in 1993. A pig in the same district was found to contain Taenia solium metacestodes (=cysticerci); the slaughtered pig body contained more than 2,000 cysticerci. In addition to morphological identification, molecular identification was also performed on the cysticerci by DNA sequencing analysis of the mitochondrial cox1 gene; they were confirmed as T. solium metacestodes. The patient is regarded as an indigenous case of neurocysticercosis infected in an endemic focus of T. solium taeniasis/cysticercosis in Oudomxay Province, Lao PDR.

Cisticercose em lingua: relato de caso / Cysticercosis in stock: case report

A cisticercose é uma doença causada pelo parasitismo da larva da tênia do porco (Cysticercus cellulosae) em tecidos humanos. É mais comum no cérebro, tecido subcutâneo, músculos e olhos. Relatamos aqui uma apresentação rara desta doença na margem lateral esquerda da língua, com diagnóstico histopatológico em biópsia excisional e retirada total da lesão. A criança acometida tinha 7 anos de idade, era do sexo masculino e previamente hígida...

Tongue cysticercosis: Case reportCysticercosis is a parasitic disease caused by the larva (Cysticercus cellulosae) of pork tapeworm. Itis more common in the brain, subcutaneous tissue, muscles and eyes. We report a rare presentation ofthis disease, affecting a previously healthy 7-year-old boy...

Diagnóstico laboratorial da neurocisticercose: revisão e perspectivas / Laboratory diagnosis of neurocysticercosis: review and perspectives

A neurocisticercose é causada por Cysticercus cellulose, a forma larval de Taenia solium, quando este se aloja no sistema nervoso central. O seu diagnóstico é realizado com base em dados clínicos, epidemiológicos, demonstração do agente etiológico pelas técnicas de imagem e testes laboratoriais. No presente estudo, apresentamos uma revisão do diagnóstico laboratorial, com ênfase no desempenho dos testes para pesquisa de anticorpos específicos e detecção de antígenos circulantes, utilização de antígeno homólogo ou heterólogo, nativo e recombinante, bem como a aplicação de métodos moleculares.

Neurocysticercosis is caused by Cysticercus cellulosae, the larval form of Taenia solium, when it lodges in the central nervous system. The diagnosis of neurocysticercosis is based on clinical and epidemiological data, neuroimaging findings of etiological agent and serologic test results. Herein we present a review of clinical diagnosis, emphasizing test performance for specific antibody and antigen detection, the use of homologous or heterologous antigen, native and recombinant antigens as well as the application of molecular methods.

Cysticercosis in experimentally and naturally infected pigs: parasitological and immunological diagnosis / Cisticercose em suínos infectados experimentalmente e naturalmente: diagnósticos parasitológico e imunológico

Our objective was to evaluate the diagnosis of swine cysticercosis by examining "ante mortem" (inspection of the tongue), "post mortem" (inspection and detailed necropsy) and ELISA for research in serum of antibodies (Ab-ELISA) and antigens (Ag-ELISA). Seven (7) pigs were experimentally infected orally with eggs of Taenia solium and another 10 were naturally infected. In the pigs experimentally infected, inspection of the tongue was negative in all animals, in the routine inspection detailed necropsy and cysticercis were identified in all of them. In pigs with heavy natural infection, inspection of the tongue identified cysticerci in two (20%), while at inspection with necropsy the parasites were identified in large quantities in all animals. In ELISA for antibody search (Ab-ELISA) TS-14 recombinant protein was used, and in search for antigen (Ag-ELISA) a monoclonal antibody against this protein. In animals experimentally infected, blood was collected weekly for 140 days. The Ab-ELISA identified an increase in titers of antibody to cysticerci 21 days after infection, and at the end of the experimental period six animals (86%) were positive to the test. The search for circulating antigens (Ag-ELISA) was positive in two pigs 28 to 91 days after infection. All naturally infected pigs were positive for Ag-ELISA and Ab-ELISA. The search for antibodies and antigens by ELISA in serum from 30 pigs of a local farm and without history of cysticercosis was negative. Thus, the use of TS-14 antigen in ELISA test (Ab-ELISA) can be useful for the diagnosis of cysticercosis in pigs with low infection.

Nosso objetivo foi avaliar o diagnóstico de cisticercose suína através do exame "ante mortem" (inspeção da língua), "post mortem" (inspeção e necropsia detalhada) e teste de ELISA para a pesquisa no soro de anticorpos (Ab-ELISA) e antígenos (Ag -ELISA). Sete (7) suínos foram infectados experimentalmente por via oral com ovos de Taenia solium e outros 10 eram portadores de infecção natural generalizada. Nos suínos experimentalmente infectados, a inspeção da língua foi negativa em todos os animais, na inspeção 4 (57%) estavam infectados, a necropsia detalhada identificou cisticercos em todos os animais. Nos animais com infecção natural generalizada, a inspeção da língua identificou cisticercos em 2 (20%), enquanto que a inspeção e a necropsia os parasitas foram identificados em grande quantidade em todos os animais. No teste de ELISA para a pesquisa de anticorpos (Ab-ELISA) foi utilizada a proteína recombinante TS-14 e para a pesquisa de antígenos (Ag-ELISA) um anticorpo monoclonal produzido contra esta proteína. Nos animais experimentalmente infectados o sangue foi coletado semanalmente por um período de 140 dias. O Ab-ELISA identificou um aumento nos títulos de anticorpos para cisticercos 21 dias após a infecção, sendo que no final do período experimental 6 animais (86%) foram positivos ao teste. A pesquisa de antígenos circulantes (Ag-ELISA), foi positiva em 2 animais, entre os dias 21 e 91 após a infecção . Todos os suínos com infecção natural generalizada foram positivos para Ag-ELISA e Ab-ELISA.A pesquisa de anticorpos e antígenos pelo ELISA realizada no soro de 30 suínos procedentes de uma criação local sem historia de cisticercose foi negativa. Assim o uso do antígeno TS-14 (Ac-ELISA), pode ser útil para o diagnóstico da cisticercose em suínos com baixa infecção.

Características clínicas y epidemiológicas de los pacientes con neurocisticercosis en un hospital general de Lima / Clinical and epidemiological features of patients with neurocysticercosis in a general hospital in Lima

Objetivo: Determinar las características clínicas y epidemiológicas de los pacientes con neurocisticercosis en un hospital general de Lima. Material y métodos: Estudio observacional, descriptivo y retrospectivo en 225 pacientes con neurocisticercosis del Hospital María Auxiliadora entre los años 2002 y 2010. Se recogió información epidemiológica y clínica de las historias clínicas. Resultados: Se confirmaron 150/225 casos. El grupo etario más afectados fue de 20 a 64 años (49,3%) y menores de 14 años (27,4%). El 52% fueron mujeres. El 90% de los pacientes procedían de Lima. El 40% fueron estudiantes, 26% amas de casa, 8,7% obreros, 10% trabajadores rurales y 5,3% desempeñaban labores domésticas. Los síntomas más frecuentes fueron cefalea intensa sin respuesta al tratamiento (76%), convulsiones (74,7%) y pérdida de conciencia (43,3%). El 68,7% de pacientes fueron hospitalizados. Todos los pacientes recibieron tratamiento médico y el 19,3% terapia quirúrgica. Conclusiones: La neurocisticercosis se observó principalmente en la población económicamente activa, comprendida entre 20 y 64 años.

Objective: To describe the clinical and epidemiological features of patients with neurocysticercosis in a general hospital in Lima. Methods: Observational and descriptive study in 225 patients with neurocysticercosis at Hospital Maria Auxiliadora from 2002 to 2010. Information was gathered from the clinical charts. Results: 150 out of 225 cases were confirmed. The two most affected age groups were patients from 20-64 years of age (49.3%) and patients below 14 years (27.4%); 52% were females; 90% come from Lima; 40% were students; 26% were housewifes; 8.7% were labors; 10% were rural workers, and 5.3% were maidservants. The most frequent symptoms were headache unresponsive to treatment (76%), seizures (74.7%) and loss of conscious (43.3%); 68.7% of patients were admitted. All patients received medical treatment and 19.3% were treated surgically. Conclusions: Neurocysticercosis was mainly observed among the active economical population from 20 to 64 years of age.