Development and evaluation of time-resolved rapid immunofluorescence test for detection of TSOL18 specific antibody in porcine cysticercosis infections.

BACKGROUND: Porcine cysticercosis, a serious zoonotic parasitic disease, is caused by the larvae of Taenia solium and has been acknowledged by the World Organization for Animal Health. The current detection methods of Cysticercus cellulosae cannot meet the needs of large-scale and rapid detection in the field. We hypothesized that the immunofluorescence chromatography test strip (ICS) for detecting Cysticercus cellulosae, according to optimization of a series of reaction systems was conducted, and sensitivity, specificity, and stability testing, and was finally compared with ELISA. This method utilizes Eu3+-labeled time-resolved fluorescent microspheres (TRFM) coupled with TSOL18 antigen to detect TSOL18 antibodies in infected pig sera. RESULTS: ICS and autopsy have highly consistent diagnostic results (n = 133), as determined by Cohen's κ analysis (κ = 0.925). And the results showed that the proposed ICS are high sensitivity (0.9459) with specificity (0.9792). The ICS was unable to detect positive samples of other parasites. It can be stored for at least six months at 4℃. CONCLUSIONS: In summary, we established a TRFM-ICS method with higher sensitivity and specificity than indirect ELISA. Results obtained from serum samples can be read within 10 min, indicating a rapid, user-friendly test suitable for large-scale field detection.

Histopathological and morphological characterization of Cysticercus fasciolaris isolated from domestic and wild rodents in Morocco.

Cysticercus fasciolaris is a parasitic helminth that usually infects feline and canine mammal hosts. The intermediate hosts (rodents, occasionally lagomorphs, and humans) get infected by the consumption of feed or water contaminated with eggs. Rodents are vectors of disease and reservoirs of various zoonotic parasites. The current survey was aimed at determining endoparasitic helminth infections in rodents in central Morocco. Sampled rodents after specific identification were sacrificed and examined to identify parasitic helminths following ethical guidelines. Parasites were identified using morphological characteristics. A total of 197 specimens of rodents were collected and examined in this study. Ten rodent species were identified morphologically as Rattus rattus, R. norvegicus, Apodemus sylvaticus, Mus musculus, M. spretus, Mastomys erythroleucus, Meriones shawi, M. libycus, Gerbillus campestris, and Lemniscomys barbarus. The parasitological results showed that metacestode of tapeworms was found encysted in the liver, the larval stage of Taenia taeniaeformis develops large multinodular fibrosarcomas which envelope the tapeworm cysts in the liver of the R. rattus and R. norvegicus. Based on morphological data, the metacestode was identified as C. fasciolaris in 23 (23/80) R. rattus 2 (2/8) and R. norvegicus with a prevalence of 11.7 % and 1.0 %, respectively. Rodents are major vectors of human and domestic animal diseases worldwide, and therefore, important parasitic zoonotic agents (C. fasciolaris), which are transmitted by black rats (R. rattus) and brown rats (R. norvegicus), must be considered to prevent the infectivity of humans, domestic animals, and livestock such as cattle, sheep, and rabbits.

Effect of Hydroxyurea on Morphology, Proliferation, and Protein Expression on Taenia crassiceps WFU Strain.

Flatworms are known for their remarkable regenerative ability, one which depends on totipotent cells known as germinative cells in cestodes. Depletion of germinative cells with hydroxyurea (HU) affects the regeneration of the parasite. Here, we studied the reduction and recovery of germinative cells in T. crassiceps cysticerci after HU treatment (25 mM and 40 mM of HU for 6 days) through in vitro assays. Viability and morphological changes were evaluated. The recovery of cysticerci's mobility and morphology was evaluated at 3 and 6 days, after 6 days of treatment. The number of proliferative cells was evaluated using EdU. Our results show morphological changes in the size, shape, and number of evaginated cysticerci at the 40 mM dose. The mobility of cysticerci was lower after 6 days of HU treatment at both concentrations. On days 3 and 6 of recovery after 25 mM of HU treatment, a partial recovery of the proliferative cells was observed. Proteomic and Gene Ontology analyses identified modifications in protein groups related to DNA binding, DNA damage, glycolytic enzymes, cytoskeleton, skeletal muscle, and RNA binding.

The muscle and neural architecture of Taenia crassiceps cysticerci revisited; implications on head-tail polarization of the larvae.

Taenia crassiceps has been used for decades as an experimental model for the study of human and porcine cysticercosis. Even though, its life cycle, tissue organization, ultrastructure and immune response elicited in the host, have been extensively described, there are many other biological questions remaining to be addressed. In the present study we revisited the muscle and neural architecture of cysticerci in two of the most frequently used strains (WFU and ORF), using conventional staining and confocal microscopy imaging, aiming to assemble an updated anatomy. Differences between both strains, including polarization processes during development of the young budding larvae, are emphasized. We also performed a search for genes that have been related to peptidergic neural processes in other related flatworms. These findings can help to understand the anatomical and molecular consequences of the scolex presence or absence in both strains.

Genetic diversity and haplotypes of Cysticercus tenuicollis isolates from slaughtered sheep and goats in Elazig and Bingol provinces of Turkey.

BACKGROUND: The cestode Taenia hydatigena uses canids, primarily dogs, as definitive hosts, whereas the metacestode larval stage cysticercus infects a range of intermediate hosts, including domestic animals such as goats, sheep and pigs. OBJECTIVES: In this study, we aimed to determine the genetic differences and haplotypes of Cysticercus tenuicollis isolates obtained from sheep and goats slaughtered in the Bingol and Elazig provinces of Turkey. METHODS: C. tenuicollis isolates were collected from 44 sheep and 26 goats slaughtered in slaughterhouses in Bingol and Elazig. After the isolation of total genomic DNA from C. tenuicollis isolates, the genetic characterization of the partial mitochondrial cytochrome oxidase 1 (CO1) gene region (866 bp) was amplified using specific primers by polymerase chain reaction (PCR), the products were then sequenced, and haplotype and genetic diversity analyses were carried out. RESULTS: As a result of the haplotype network analyses, 34 different haplotypes were detected around the main haplotype (Hap02) arranged in a star-like configuration and separated from other haplotypes by 1-28 mutation steps and covering 22.85% (16/70) of all isolates. Twenty-seven polymorphic fields were detected, 77.77% (21/27) of which were parsimony-informative, and secondary haplotype and nucleotide diversity were observed. Additionally, we detected high intraspecies haplotype diversity (hd: 0.933) and high nucleotide diversity (π: 0.00383), with 27 different nucleotide variation positions among the haplotypes of the isolates. Tajima's D value was negative, indicating population expansion and/or selection purification. The significantly negative Fu's Fs values indicated recent population expansion or the presence of expected rare haplotypes. CONCLUSION: The results of this study confirmed that C. tenuicollis isolates clustered in one lineage and were closely related to the relevant reference sequences in different countries, confirming the circulation of C. tenuicollis in different geographical regions.

Concordance between two monoclonal antibody-based antigen detection enzyme-linked immunosorbent assays for measuring cysticercal antigen levels in sera from pigs experimentally infected with Taenia solium and Taenia hydatigena.

BACKGROUND: Antigen detection in Taenia solium cysticercosis confirms viable infection in the intermediate host (either pig or human). The reference B158/B60 monoclonal antibody (mAb)-based Ag-enzyme-linked immunosorbent assay (ELISA) has acceptable levels of sensitivity and specificity in human neurocysticercosis with multiple brain cysts, although its sensitivity is lower in cases with single brain cysts, whereas in porcine cysticercosis the assay specificity is affected by its frequent cross-reaction with Taenia hydatigena, another common cestode found in pigs. Our group has produced 21 anti-T. solium mAbs reacting against antigens of the whole cyst, vesicular fluid, and secretory/excretory products, identifying TsW8/TsW5 as the most promising pair of mAbs for an Ag-ELISA. METHODS: We report the use of the TsW8/TsW5 Ag-ELISA to measure cysticercus antigen levels [expressed as optical density (OD) values] in two panels of sera collected from day 0 (baseline) to day 90 postinfection (PI) from pigs experimentally infected with T. solium (n = 26) and T. hydatigena (n = 12). At baseline and on days 28 and 90 PI, we used Bland-Altman (BA) analysis and Lin's concordance correlation coefficients (CCC) to determine the concordance between the TsW8/TsW5 and the B158/B60 Ag-ELISA. RESULTS: The TsW8/TsW5 Ag-ELISA was able to efficiently measure circulating antigen levels in T. solium-infected pigs, similar to that obtained with the B158/B60 Ag-ELISA. Almost all paired log-OD differences between assays were within the limits of agreement (LoA) in the BA analysis at baseline and on days 28 and 90 PI (92.3%, 100%, and 100%, respectively), and a high concordance of log-ODs between assays was also found (Lin's CCC: 0.69, 0.92, and 0.96, respectively, all P < 0.001). In pigs infected with T. hydatigena, almost all paired log-OD differences were within the LoA in the BA analysis, whereas the concordance of log-ODs between assays was low at baseline (Lin's CCC: 0.24) but increased on days 28 and 90 PI (Lins' CCC: 0.88 and 0.98, P < 0.001). CONCLUSIONS/SIGNIFICANCE: The TsW8/TsW5 Ag-ELISA recognizes antigens in pigs with T. solium cysticercosis and is highly concordant with the B158/B60 Ag-ELISA. However, its diagnostic use is hampered by cross-reactions with T. hydatigena, as in other mAb-based Ag-ELISAs.

Using the model cestode Taenia crassiceps for the study of cysticercosis.

Taenia solium is the aetiological agent of taeniasis/cysticercosis, one of the most severe neglected tropical diseases (NTD) according to the World Health Organization (WHO). The life cycle of T. solium alternates between pigs (intermediate host) and humans (definitive host). In addition, humans can act as accidental intermediate hosts if they ingest infective eggs. In this case, the most severe condition of the disease occurs when parasites invade the central nervous system, causing neurocysticercosis (NCC). The complexity of the life cycle of T. solium imposes a barrier to study this pathogen thoroughly. Thus, related species, such as T. crassiceps are commonly used. Due to its capacity to multiply asexually, T. crassiceps can be maintained by serial passage in laboratory mice in standard biosecurity level facilities. In addition, an in vitro system to generate cysticerci in the presence of feeder cells has been recently developed. Despite model species display biological differences with their zoonotic counterparts, they have historically helped to understand the biology of the related pathogenic species and hence, generate improvements in NTD detection and control. In this chapter, we describe the procedures to carry out both in vivo and in vitro systems for T. crassiceps in the laboratory.

The infection of Cysticercus fasciolaris in natural rats (Rattus species) residing in human residence areas, Nakhon Si Thammarat, Thailand.

Cysticercus fasciolaris (C. fasciolaris) is the larval stage of a cestode parasite named Taenia taeniaeformis (T. taeniaeformis). C. fasiolaris is found in small rodents, especially rats. Rattus species are listed as intermediate hosts of this parasite, and cats are the main definitive host of C. fasiolaris. The objective of this study was to study the pathological, microscopic, and molecular aspects of C. fasciolaris in rodents residing in human residence areas. One hundred and two rodents were trapped in human settlements and dissected for larva-containing cyst examinations in the body cavity. The larvae of C. fasciolaris were investigated using histopathological examination, microscopic observations under a stereomicroscope and scanning electron microscope, and molecular detection using polymerase chain reaction. The prevalence of hepatic cysts containing larvae was 8.91% (95% CI = 4.16-16.24). In addition, the older larvae also had longer micropapillae. Histopathological investigation revealed normal hepatic tissue containing larvae and a scanty fluid cyst. The cyst capsule contains mostly mononuclear cells and spindle cells in all infected rats. The molecular detection using two primer sets revealed the amplicons were similar to the clade of C. fasciolaris. In the future, more investigation is necessary to fully understand the parasite's molecular pathogenesis and virulent molecules, which are less obvious.

A rare case of Cysticercus tenuicollis infection in a neonate lamb: Evidence of prenatal transmission.

Cysticercosis develops in lambs following a Cysticercus tenuicollis infestation, which is the larval stage of Taenia hydatigena. A 7-day-old lamb was examined for depression, anorexia, fever (40.5°C), congested mucus membranes, reluctance to move, and a hunched back. Upon necropsy, congestion was noted in the intestines and brain, and the heart had a loose consistency. Soft and pulpy kidneys were evident coupled with watery intestinal contents. Epsilon toxin (Clostridium perfringens type D toxin) was detected using enzyme-linked immunosorbent assay. A transparent cystic structure was incidentally found attached to the pancreas, within which a scolex was well demonstrated upon histopathology. Chronic active peritonitis was diagnosed at the cyst attachment site. C. tenuicollis was confirmed by polymerase chain reaction and genome sequencing. This report describes prenatal transmission of C. tenuicollis in the present lamb, although this condition is quite rare.

[Prevalence of taeniasis and sero - prevalence of anti - cysticercus antibody among residents in Tibetan agricultural areas of Sichuan Province].

OBJECTIVE: To investigate the prevalence and epidemiological characteristics of taeniasis and cysticercosis among residents in Tibetan agricultural areas of Sichuan Province, so as to provide insights for the prevention and control of taeniasis and cysticercosis. METHODS: From 2016 to 2022, Kangding City, Daocheng County, Derong County, Ruoergai County and Muli Tibetan Autonomous County were sampled from Tibetan agricultural areas of Ganzi Tibetan Autonomous Prefecture, Aba Tibetan and Qiang Autonomous Prefecture and Liangshan Yi Autonomous Prefecture in Sichuan Province, and 1 to 6 townships were sampled from each county (district), followed by 4 to 7 villages sampled from each township. Primary school children were sampled using a cluster sampling method, and permanent residents at ages of over 16 years were randomly sampled from each village. Participants' demographics, history of tapeworm excretion during the past year and clinical symptoms and signs of cysticercosis were collected through questionnaire surveys, and participants' stool and venous blood samples were collected. Taenia eggs were detected in stool samples using the direct smear method, and deworming was performed among taeniasis patients with areca nut-squash seeds. The tapeworm species were identified using a multiplex PCR assay, and serum specific IgG antibody against cysticercus was detected using enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 5 249 respondents participated in the questionnaire survey, including 603 respondents (11.5%) with a self-reported history of proglottids secretion during the past year. A total of 3 976 residents were subjected to stool examinations, and the detection of Taenia eggs was 6.5%. Of 258 participants undergoing deworming, there were 403 cases (94.2%) with excretions of Taenia worms or proglottids. The mean prevalence of taeniasis was 10.9% (439/4 043), and there were gender-, age- and region-specific prevalence rates of taeniasis (χ2 = 36.73, 126.31 and 163.41, all P values < 0.05). Multiplex PCR assays detected 41 cases with T. solium infections (12.5%), 197 cases with T. saginata infections (59.9%) and 91 cases with T. asiatica infections (27.6%) among 329 patients undergoing deworming, and there were region-specific prevalence rates of T. solium, T. saginata and T. asiatica infections (χ2 = 45.39, P < 0.05). In addition, the sero-prevalence of anti-cysticercus IgG antibody was 7.0% (345/4 933), and there were age- and region-specific sero-prevalence rates of anti-cysticercus IgG antibody (χ2 = 13.49 and 51.76, both P values < 0.05). CONCLUSIONS: Multiple Taenia species are prevalent in Tibetan agricultural areas of Sichuan Province and the sero-prevalence of anti-cysticercus antibody is high among residents. Monitoring and control of taeniasis and cysticercosis should be strengthened.

Nexus between prevalence of Taenia solium (Cysticercus cellulosae) in pigs and retrospective study of humans cysticercosis cases in Benin hospitals.

The cysticercosis is a major zoonotic disease in many developing countries. This study was carried out to determine the prevalence of porcine cysticercosis and to identify potential risk factors associated and their seroprevalence as well in Benin. The tongue palpation was performed on 4150 pigs from the pig farmers were randomly selected to diagnose cysticercosis and to assess the risk factors associated. In slaughterhouses, 792 pigs were inspected. Serum samples from 460 pigs were tested of the presence of Taenia solium cysticercosis using the HP10 antigen-ELISA (Ag-ELISA). A logistic regression model with a 95% confidence interval was used to analyse the main risk factors associated. The tongue palpation revealed a prevalence of 4.79% at the level of slaughterhouses versus 7.82% in the post-mortem inspection. The Ag-ELISA revealed that more than 40% of the pigs were positive. The risk factors identified were the origin area of pigs, the history of cysticercosis in the farms, the presence or absence of a latrine, the pig-farming systems, the age class of pigs and the source of watering. In addition, 34.48% individuals was tested for taenia versus 13.79% for cysticercosis in the hospitals surveyed. Our results showed the major revelation of porcine cysticercosis and its very high level of risk for a public health in all departments of Benin. Therefore, this study suggests the need to continue epidemiological and sociological surveys with a "One Health" approach in order to design and implement effective control measures in this country of high consumption of pigs.

Serological surveillance of parasitic zoonoses in 13 highlands regions of Peru: Period 2016-2019. / Vigilancia serológica de la zoonosis parasitaria en 13 regiones de la sierra del Perú: Periodo 2016-2019.

OBJECTIVES.: Motivation for the study. To understand the characteristics and distribution of the main parasitic zoonoses in Peru and to generate data for decision making in surveillance, prevention and control. Main findings. These parasitic zoonoses are distributed in areas of extreme poverty in the central and southern highlands of Peru. Fascioliasis seropositivity was found to be higher than for echinococcosis and cysticercosis. In addition, sociodemographic characteristics and lifestyle habits influence the transmission of these zoonoses. Implications. An active search for these zoonoses should be carried out in other risk areas with similar epidemiological characteristics to determine the prevalence of each of these zoonoses and implement multisectoral prevention and control programs. . To determine seropositivity to anti-IgG antibodies against Echinococcus granulosus, Fasciola hepatica and Taenia solium cysticercus infection and to describe the characteristics of the infected patients in 13 regions of the Peruvian highlands between 2016 and 2019. MATERIALS AND METHODS.: Cross-sectional, observational study, in which we analyzed 7811 epidemiological records of laboratory-based surveillance of parasitic zoonoses from 2016 to 2019. Diagnosis was established by detecting IgG type anti-E. granulosus, F. hepatica and T. solium cysticercus antibodies using native antigens by enzyme-linked immunosorbent assay (ELISA) and Immunoblot. We evaluated the difference in the frequency of the cases according to identified characteristics using Pearson's chi-square test and Fisher's exact test. RESULTS.: Seropositivity was 7.9% for fascioliasis, 4.9% for cystic echinococcosis, and 2.3% for T. solium cysticercus. These rates were higher in Cerro de Pasco for cystic echinococcosis (24.5%), in Ayacucho for T. solium cysticercus (4.5%) and in Puno for fascioliasis (40.6%). Regarding the sociodemographic characteristics, we found a statistically significant difference in the frequency of cases for all zoonoses according to age group, occupation, and region of residence. We also found a difference with the consumption of vegetables in emollients, and between clinical-epidemiological characteristics and having a family history of parasitic zoonoses. CONCLUSIONS.: From the 7811 samples, we found that these parasitic zoonoses are distributed in 13 regions of the Peruvian highlands, and represent a major health problem, with frequencies that change according to different characteristics.

OBJETIVOS.: Determinar la seropositividad a anticuerpos anti-IgG por infección de Echinococcus granulosus, Fasciola hepatica y cisticerco de Taenia solium y describir las características de los infectados en 13 regiones de la sierra peruana entre 2016 y 2019. MATERIALES Y MÉTODOS.: Estudio observacional transversal, que analizó 7811 fichas epidemiológicas de la vigilancia basada en laboratorio de las zoonosis parasitarias del periodo 2016-2019. El diagnóstico se realizó mediante la detección de anticuerpos tipo IgG anti E. granulosus, F. hepatica y cisticerco de T. solium utilizando antígenos nativos mediante el ensayo inmunoabsorbente ligado a enzimas (ELISA) e Inmunoblot. La diferencia en la frecuencia de casos de estas zoonosis según características identificadas se realizó mediante la prueba chi-cuadrado de Pearson y prueba exacta de Fisher. RESULTADOS.: Se determinó una seropositividad de 7,9% para fascioliasis, 4,9% para equinococosis quística, y 2,3% para cisticerco de T. solium. Estas frecuencias fueron mayores en Cerro de Pasco para equinococosis quística (24,5%), en Ayacucho para cisticerco de T. solium (4,5%) y en Puno para fascioliasis (40,6%). Entre las características sociodemográficas, se encontró una diferencia estadísticamente significativa en la frecuencia de casos para todas las zoonosis según grupo etario, ocupación, y región de residencia. Además, se encontró diferencia con el consumo de verduras en emolientes, y entre las características clínico-epidemiológicas con tener antecedentes familiares de las zoonosis parasitarias. CONCLUSIONES.: A partir de las 7811 muestras evaluadas, se encontró que estas zoonosis parasitarias están distribuidas en 13 regiones de la sierra del Perú, ocasionando un problema de salud importante, con frecuencias que varían según diversas características.

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)

Oral cysticercosis in the paediatric patient: report of six cases / Cisticercosis oral en el paciente pediátrico: informe de seis casos

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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.