Changes in inflammatory gene expression in brain tissue adjacent and distant to a viable cyst in a rat model for neurocysticercosis.

BACKGROUND: The parasite Taenia solium causes neurocysticercosis (NCC) in humans and is a common cause of adult-onset epilepsy in the developing world. Hippocampal atrophy, which occurs far from the cyst, is an emerging new complication of NCC. Evaluation of molecular pathways in brain regions close to and distant from the cyst could offer insight into this pathology. METHODS: Rats were inoculated intracranially with T. solium oncospheres. After 4 months, RNA was extracted from brain tissue samples in rats with NCC and uninfected controls, and cDNA was generated. Expression of 38 genes related to different molecular pathways involved in the inflammatory response and healing was assessed by RT-PCR array. RESULTS: Inflammatory cytokines IFN-γ, TNF-α, and IL-1, together with TGF-ß and ARG-1, were overexpressed in tissue close to the parasite compared to non-infected tissue. Genes for IL-1A, CSF-1, FN-1, COL-3A1, and MMP-2 were overexpressed in contralateral tissue compared to non-infected tissue. CONCLUSIONS: The viable cysticerci in the rat model for NCC is characterized by increased expression of genes associated with a proinflammatory response and fibrosis-related proteins, which may mediate the chronic state of infection. These pathways appear to influence regions far from the cyst, which may explain the emerging association between NCC and hippocampal atrophy.

Cerebral Coenurosis of a Long-Tailed Goral, Naemorhedus caudatus, in Korea.

We intended to describe a case of cerebral coenurosis in a long-tailed goral, Naemorhedus caudatus, from Hwacheon-gun, Gangwon-do (Province), in the Korea. The goral, a 10-year-old male, was suffering from neurological symptoms, such as turning the circle to one side without lifting the head straight, and died at 30 days after admission to the wildlife medical rescue center in Chuncheon-si, Gangwon-do. A fluid-filled cyst was detected in the left cerebral hemisphere by computed tomography and magnetic resonance imaging. The cyst removed from the deceased goral was transparent, about 3×3 cm in size, contained a clear fluid and approximately 320 protoscolices invaginating from the internal germinal layer. The protoscolex had 4 suckers and a rostellum with 28 hooklets arranged in 2 rows. By the present study, a case of cerebral coenurosis was first confirmed in a long-tailed goral, N. caudatus, from Gangwon-do, in Korea. The residents frequently exposed in the sylvatic environment should be careful the accidental infections of zoonotic metacestode of Taenia multiceps, Coenurus cerebralis, in Korea.

Zoonotic Taenia infections with focus on cysticercosis due to Taenia solium in swine and humans.

Zoonotic taeniasis caused by the adult stage of Taenia solium, Taenia saginata or Taenia asiatica are considered neglected tropical diseases by the World Health Organization. The life cycle of these 3 metazoan species is very similar and includes an intermediate host: pigs in the case of T. solium and T. asiatica, and cattle in the case of T. saginata. By eating meat (pork/T. solium, T. asiatica; beef/T. saginata) containing live cysticerci, humans develop taeniasis, which is practically asymptomatic but is the main risk factor for intermediate hosts to become infected. T. saginata causes bovine cysticercosis, while T. solium and T. asiatica cause swine cysticercosis, of veterinary and economic importance. T. solium cysticerci cause neurological disease in humans: neurocysticercosis. Cysticerci develop after ingesting microscopic eggs released from a human tapeworm carrier. Here we describe the life stages of the parasites, diagnosis, pathogenesis, symptomatology of neurocysticercosis, and prevention and control measures. Highlighting the need to validate diagnostic tools, treatments and vaccination in endemic areas, with the challenge of addressing the most vulnerable populations that lack resources. If people understand the transmission route, avoid eating uncooked or insufficiently cooked meat and have adequate hygienic habits, the life cycle of the 3 zoonotic Taenia species may be interrupted. In addition, we describe the growing field of immune response and immunomodulation elicited by the parasites, which may provide essential tools for diagnosis, treatment, control of taeniasis/cysticercosis, as well as for identification of parasite-derived immunomodulators that could aid in the treatment of emerging inflammatory diseases worldwide.

Performance of Ag-ELISA in the diagnosis of Taenia solium cysticercosis in naturally infected pigs in Tanzania.

BACKGROUND: Taenia solium is a zoonotic parasite responsible for neurocysticercosis-a major cause of late-onset acquired epilepsy in humans. Lack of affordable, specific and sensitive diagnostic tools hampers control of the parasite. This study assessed the performance of an antigen detection enzyme-linked immunosorbent assay (Ag-ELISA) in the diagnosis of viable T. solium cysticercosis in naturally infected slaughter-age pigs in an endemic area in Tanzania. METHODS: A total of 350 pigs were bled before they were slaughtered and their carcases examined. Serum was analyzed for circulating antigens by using a monoclonal antibody-based B158/B60 Ag-ELISA. Each carcase was examined for the presence of Taenia hydatigena cysticerci and half carcase musculature together with the whole brain, head muscles, tongue, heart and diaphragm were sliced with fine cuts (< 0.5 cm) to reveal and enumerate T. solium cysticerci. Half carcase dissection can detect at least 84% of infected pigs. Prevalence and their 95% confidence intervals (CI) were calculated in Stata 12. Sensitivity, specificity, predictive values and likelihood ratios were determined. RESULTS: Twenty-nine pigs (8.3%, 95% CI: 5.6-11.7%) had viable T. solium cysticerci while 11 pigs had T. hydatigena cysticerci (3.1%, 95% CI: 1.6-5.5%). No co-infection was observed. Sixty-eight pigs (19.4%, 95% CI: 15.4-20%) tested positive on Ag-ELISA; of these, 24 had T. solium cysticerci and 7 had T. hydatigena cysticerci. Sensitivity and specificity were determined to be 82.7% and 86.3%, respectively. Positive and negative predictive values were 35.2% and 98.2%, respectively. Likelihood ratios for positive and negative Ag-ELISA test results were 6.0 and 0.2, respectively. There was a significant positive correlation between the titre of circulating antigens and intensity of T. solium cysticerci (r(348) = 0.63, P < 0.001). CONCLUSIONS: The Ag-ELISA test characteristics reported in this study indicate that the test is more reliable in ruling out T. solium cysticercosis in pigs, than in confirming it. Hence, a negative result will almost certainly indicate that a pig has no infection, but a positive result should always be interpreted with caution. Estimates of T. solium prevalence based on Ag-ELISA results should, therefore, be adjusted for test performance characteristics and occurrence of T. hydatigena.

Taenia solium proteins: a beautiful kaleidoscope of pro and anti-inflammatory antigens.

Background: Neurocysticercosis (NCC) is an acquired infection of central nervous system associated with epileptic seizures. The parasite 'Taenia solium' causes this disease and has a complex life cycle and molts into various stages that influence the host-parasite interaction. The disease has a long asymptomatic phase with viable cyst and degeneration of cyst and leaking cyst fluid has been associated with symptomatic phase. The parasite proteome holds the answers and clues to this complex clinical presentation and hence unraveling of proteome of parasite antigens is needed for better understanding of host-parasite interactions. Objective: To understand the proteome make-up of T. solium cyst vesicular fluid (VF) and excretory secretory proteins (ESPs). Methodology: The VF and ESPs for the study were prepared from cyst harvested from naturally infected swine. The samples were prepared for nano LC-MS by in-tube digestion of proteins. The spectra obtained were annotated and enrichment analysis was performed and in silico analysis was done. Results:T. solium VF and ESPs have 206 and 247 proteins of varied make-up including pro-inflammatory and anti-inflammatory nature. Conclusions: Due to varied make-up of VF and ESPs it can generate complex humoral and cellular immune response.

Coenurosis of Yak, Bos grunniens, caused by Taenia multiceps: A Case Report with Molecular Identification in Qinghai Tibetan Plateau Area, China.

Coenurosis is an important zoonotic helminthic disease caused by the larval stage of the tapeworm Taenia multiceps. This parasite typically infects the brain of the intermediate hosts, including sheep, goat, cattle and even humans. We report a case of T. multiceps infection in a yak confirmed by clinical symptoms, morphological characteristics, and molecular and phylogenetic analyses. The coenurus was thin-walled, whitish, and spherical in shape with a diameter of 10 cm. The parasite species was identified as T. multiceps by PCR amplification and sequencing of the 18S rRNA, cox1 and nad1 genes. Three gene sequences all showed high homology (all above 97%) with the reference sequences from different hosts. Moreover, phylogenetic reconstructions with the 3 published Taenia gene sequences confirmed that the Qinghai yak isolate was closely related to T. multiceps. Although there are advanced diagnosis and treatment methods for coenurosis, early infection is difficult to diagnose. Importantly, the findings of yak infection case should not be ignored due to its zoonotic potential.

First report of an outbreak of cerebral coenurosis in Dhofari goats in Oman.

This study documented the first outbreak of cerebral coenurosis in goats in Salalah, southern Oman. Deaths of 130 (16.6%) adult native goats in a herd (n=780) were reported from January to June 2017. Affected goats showed various nervous signs ended by death. Investigations for thiamine deficiency, polioencephalomalacia, caprine arthritis encephalitis, and listeriosis were negative. Upon necropsy, multiple (1-4) thin-walled cysts 2-3.5 cm in diameter containing clear fluid with numerous clusters of protoscolices in the cerebrum and cerebellum had replaced the brain parenchyma, causing space-occupying lesions. Parasitologically, the recovered cysts were Coenurus cerebralis, based on the arrangement of protoscolices, and the number and size of their hooks. Morphologically, each protoscolex had four suckers and a rostellum with double-crown hooks. The large and small hooks were 157.7±0.5 µm and 115±0.6 µm in length, respectively. Histopathologically, the parasite destroyed the affected tissues associated with multifocal to diffuse lymphocytic, non-suppurative meningoencephalitis; ischemic neuronal necrosis; and malacia. This is the first report of cerebral coenurosis in livestock in Oman, which should alert the local public health authorities for the application of prevention and control measures.

First report of an outbreak of cerebral coenurosis in Dhofari goats in Oman / Primeiro relato de um surto de coenurose cerebral em cabras Dhofari em Omã

Abstract This study documented the first outbreak of cerebral coenurosis in goats in Salalah, southern Oman. Deaths of 130 (16.6%) adult native goats in a herd (n=780) were reported from January to June 2017. Affected goats showed various nervous signs ended by death. Investigations for thiamine deficiency, polioencephalomalacia, caprine arthritis encephalitis, and listeriosis were negative. Upon necropsy, multiple (1-4) thin-walled cysts 2-3.5 cm in diameter containing clear fluid with numerous clusters of protoscolices in the cerebrum and cerebellum had replaced the brain parenchyma, causing space-occupying lesions. Parasitologically, the recovered cysts were Coenurus cerebralis, based on the arrangement of protoscolices, and the number and size of their hooks. Morphologically, each protoscolex had four suckers and a rostellum with double-crown hooks. The large and small hooks were 157.7±0.5 µm and 115±0.6 µm in length, respectively. Histopathologically, the parasite destroyed the affected tissues associated with multifocal to diffuse lymphocytic, non-suppurative meningoencephalitis; ischemic neuronal necrosis; and malacia. This is the first report of cerebral coenurosis in livestock in Oman, which should alert the local public health authorities for the application of prevention and control measures.

Resumo Este estudo documentou o primeiro surto de coenurose cerebral em cabras em Salalah, Oman. A morte de 130 (16,6%) caprinos adultos nativos (n=780) foi relatada de janeiro a junho de 2017. As cabras afetadas mostraram distúrbios neurológicos, que culminaram em óbito. Investigações para deficiência de tiamina, polioencefalomalácia, encefalite por artrite caprina e listeriose foram negativas. Na necropsia, múltiplos (1-4) cistos de paredes finas com 2-3,5 cm de diâmetro contendo líquido claro com numerosos aglomerados de protoescólices no cérebro e no cerebelo haviam substituído o parênquima cerebral, causando compressão nas estruturas adjacentes. Os cistos recuperados foram identificados como sendo de Coenurus cerebralis, com base no arranjo dos protoescólices, e no número e tamanho de seus ganchos. Morfologicamente, cada protoescólice tinha quatro ventosas e um rostelo com dupla coroa de ganchos. Os ganchos grandes e pequenos tinham 157,7±0,5 µm e 115±0,6 µm de comprimento, respectivamente. Histopatologicamente, o parasita causou a destruição dos tecidos afetados associada à meningoencefalite linfocítica não-supurativa, que variou de multifocal a difusa, necrose neuronal isquêmica e malacia. Este é o primeiro relato de coenurose em ruminantes no Oman, o que deve servir de alerta para as autoridades locais da área de saúde para a aplicação de medidas de prevenção e controle.

TNF-α blockade suppresses pericystic inflammation following anthelmintic treatment in porcine neurocysticercosis.

BACKGROUND: Neurocysticercosis (NCC) is an infection of the brain with the larval cyst of the tapeworm, Taenia solium. Cysticidal treatment induces parasite killing resulting in a post inflammatory response and seizures, which generally requires corticosteroid treatment to control inflammation. The nature of this response and how to best control it is unclear. We investigated the anti-inflammatory effects of pretreatment with etanercept (ETN), an anti-tumor necrosis factor agent, or dexamethasone (DEX), a high potency corticosteroid, on the post treatment inflammatory response in naturally infected pigs with neurocysticercosis after a single dose of the cysticidal drug praziquantel (PZQ). METHODOLOGY/PRINCIPAL FINDINGS: We followed the methods from a previously developed treatment model of NCC in naturally infected swine. The four study groups of infected pigs included 3 groups treated with PZQ on day 0: PZQ-treated alone (100 mg/kg PO; n = 9), pretreated with dexamethasone (DEX, 0.2 mg/kg IM administered on days -1, +1 and +3; n = 6), and pretreated with etanercept (ETN, 25 mg IM per animal on days -7 and 0; n = 6). The fourth group remained untreated (n = 3). As measured by quantitative RT-PCR, ETN pretreatment depressed transcription of a wide range of proinflammatory, regulatory and matrix protease encoding genes at 120 hr post PZQ treatment in capsules of cysts that demonstrated extravasated Evans Blue (EB) (a measure of blood brain barrier dysfunction) compared to animals not receiving ETN. Transcription was significantly depressed for the proinflammatory genes tumor necrosis factor (TNF)-α, and interferon (IFN)-γ; the inflammation regulating genes cytotoxic T-lymphocyte-associated protein (CTLA)4, interleukin (IL)-13 and transforming growth factor (TGF)-ß; the tissue remodeling genes matrix metalloprotease (MMP)1 and 9, tissue inhibitors of metalloproteases (TIMP)1 and 2, and the genes regulating endothelial function vascular endothelial growth factor (VEGF)1, angiopoietin (Ang)1, Ang 2, and platelet endothelial cell adhesion molecule (PECAM)-1. In contrast, transcription was only modestly decreased in the DEX pretreated pigs compared to PZQ alone, and only for TNF-α, IL-6, IFN-γ, TGF-ß and Ang1. IL-10 was not affected by either ETN or DEX pretreatments. The degree of inflammation, assessed by semi-quantitative inflammatory scores, was modestly decreased in both ETN and DEX pretreated animals compared to PZQ treated pigs whereas cyst damage scores were moderately decreased only in cysts from DEX pretreated pigs. However, the proportion of cysts with EB extravasation was not significantly changed in ETN and DEX pretreated groups. CONCLUSIONS/SIGNIFICANCE: Overall, TNF-α blockade using ETN treatment modulated expression of a large variety of genes that play a role in induction and control of inflammation and structural changes. In contrast the number of inflammatory cells was only moderately decreased suggesting weaker effects on cell migration into the inflammatory capsules surrounding cysts than on release of modulatory molecules. Taken together, these data suggest that TNF-α blockade may provide a viable strategy to manage post-treatment pericystic inflammation that follows antiparasitic therapy for neurocysticercosis.

Fasciclin-calcareous corpuscle binary complex mediated protein-protein interactions in Taenia solium metacestode.

BACKGROUND: Neurocysticercosis (NC) caused by Taenia solium metacestode (TsM) is a serious neurological disease of global concern. Diverse bioactive molecules involved in the long-term survival of TsM might contribute to disease progression. Fasciclin (Fas) is an extracellular protein that mediates adhesion, migration and differentiation of cells by interacting with other molecules. We hypothesized that TsMFas might bind to calcareous corpuscle (CC) through its adhesive property and participate in crucial protein-protein interactions, thus contributing to the creation of a symbiotic interactome network. METHODS: Two paralogous TsMFas (TsMFas1 and TsMFas2) were isolated, and their molecular properties were characterized. The co-localization pattern of TsMFas1 and TsMFas2 with CC was determined. CC-TsMFas binary complex was generated by incubating CC with recombinant proteins (rTsMFas1 and 2). In vitro binding assay of CC-rTsMFas1 or CC-rTsMFas2 binary complex with TsM cellular proteins extracted from scolex and neck was conducted. Their binding partners were identified through proteomic analysis. Integrated protein-protein interaction networks were established. RESULTS: TsMFas1 (6072 bp long) was composed of 15 exons (841 amino acid polypeptide) interrupted by 14 introns. TsMFas2 (5201 bp long) comprised of 11 exons (597 amino acids) and 10 intervening introns. These proteins displayed 22% amino acid sequence identity to each other, but tightly conserved Fas-related domains. Several isoforms of Fas1 and Fas2 proteins might have been expressed through post-translational modifications. They showed adhesion activity with other cells. TsMFas proteins were largely distributed in parenchymal regions of the scolex and bladder wall. These molecules were co-localized with CC, a unique organelle found in platyhelminths. Subsequent proteome analysis of CC-Fas binary complex mediated protein-protein interactions revealed seven protein ligands in the TsM cellular proteins. Their functions were mainly segregated into carbohydrate metabolism (enolase, phosphoenolpyruvate carboxykinase, phosphoglycerate kinase and glyceraldehyde 3-phosphate dehydrogenase) and cytoskeleton/cellular motility (actin, paramyosin and innexin nuc-9). Those proteins had direct (physical) and/or indirect (functional) relationships along with their biochemical properties and biological roles. CONCLUSION: Protein repertoires strongly suggest that TsMFas and CC may symbiotically mediate protein-protein interactions during biological processes to maintain efficacious homeostatic functions and ensure the prolonged survival of TsM in the host.

Adhesion molecules, chemokines and matrix metallo-proteinases response after albendazole and albendazole plus steroid therapy in swine neurocysticercosis.

The treatment of neurocysticercosis (NCC) varies with location, number and stage of the Taenia solium cysticerci (cysts). Albendazole (ABZ) effectively kills cysticerci, and subsequently induces neuro-inflammation facilitated by leukocyte infiltration. We hypothesize that immune response varies around drug responder (degenerating/dying) and non-responder (viable) cysts after ABZ and ABZ plus steroid (ABZS) therapy, which may determine the disease pathogenesis. Twenty cysticercotic swine were treated with ABZ (n = 10; group1) and ABZS (n = 10; group2). Expression of adhesion molecules, chemokines and matrix metallo-proteinases (MMPs) was measured by qRT-PCR (quantitative reverse transcriptase-polymerase chain reaction) and ELISA. Gelatin gel zymography was performed to detect the activity of MMP-2 and -9. In group1, ABZ therapy induced higher expressions of ICAM-1 (intercellular adhesion molecule-1), VCAM-1 (vascular cell adhesion molecule-1), E-selectin, MCP-1 (monocyte chemotactic protein-1), Eotaxin-1, MIP-1α (macrophage inflammatory protein-1α), RANTES (regulated on activation, normal T cell expressed and secreted), MMP-2 and MMP-9 around ABZ responder (AR) cysts. Three pigs with cyst burdens ≥10 died following ABZ therapy. However, in group2, moderate expressions of ICAM-1, VCAM-1, E-selectin, RANTES and MMP-9 were associated with ABZS responder (ASR), whereas low expressions of these molecules were associated with ABZS non-responder (ASNR) cysts. In conclusion, ABZ alone therapy is not safe since it causes death of pigs due to higher inflammatory immune response around dying cysts. However, combination therapy is an effective treatment regimen even with the high cyst burden.

Radiological evolution of porcine neurocysticercosis after combined antiparasitic treatment with praziquantel and albendazole.

BACKGROUND: The onset of anthelmintic treatment of neurocysticercosis (NCC) provokes an acute immune response of the host, which in human cases is associated with exacerbation of neurological symptoms. This inflammation can occur at the first days of therapy. So, changes in the brain cysts appearance may be detected by medical imaging. We evaluated radiological changes in the appearance of brain cysts (enhancement and size) on days two and five after the onset of antiparasitic treatment using naturally infected pigs as a model for human NCC. METHODS AND RESULTS: Contrast T1-weighted magnetic resonance imaging with gadolinium was performed before and after antiparasitic treatment. Eight NCC-infected pigs were treated with praziquantel plus albendazole and euthanized two (n = 4) and five (n = 4) days after treatment; another group of four infected pigs served as untreated controls. For each lesion, gadolinium enhancement intensity (GEI) and cyst volume were measured at baseline and after antiparasitic treatment. Volume and GEI quantification ratios (post/pre-treatment measures) were used to appraise the effect of treatment. Cysts from untreated pigs showed little variations between their basal and post treatment measures. At days 2 and 5 there were significant increases in GEI ratio compared with the untreated group (1.32 and 1.47 vs 1.01, p = 0.021 and p = 0.021). Cyst volume ratios were significantly lower at days 2 and 5 compared with the untreated group (0.60 and 0.22 vs 0.95, p = 0.04 and p = 0.02). Cysts with lower cyst volume ratios showed more marked post-treatment inflammation, loss of vesicular fluid and cyst wall wrinkling. CONCLUSION/SIGNIFICANCE: A significant and drastic reduction of cyst size and increased pericystic enhancement occur in the initial days after antiparasitic treatment as an effect of acute perilesional immune response. These significant changes showed that early anthelmintic efficacy (day two) can be detected using magnetic resonance imaging.

Clinical, pathological and molecular evaluations and CT scan screening of coenurosis (Coenurus cerebralis) in sheep and calves / Avaliação clínica, patológica e molecular e tomografia computadorizada de cenurose (Coenurus cerebralis) em carneiros e bezerros

Abstract The aims of this study were to diagnose coenurosis by means of computerized tomography (CT) scan imaging and molecular characterization of the CO1 gene using the polymerase chain reaction (PCR). Sheep and calves were necropsied, and CT scans on the cephalic region were performed on the animals. Sections of brain tissue infected with parasites were then stained with hematoxylin and eosin for microscopic examination. Material collected from brain cysts was fixed in 70% ethanol. PCR amplification was carried out using the CO1 mitochondrial gene. A total of 60 calves and 80 sheep were examined clinically and, of these, 15 calves and 38 sheep showed signs of depression, with counterclockwise circling movements and altered head carriage. Four sheep and one calf were necropsied, and C. cerebralis cysts were detected in all of them. A hypodense cyst was monitored in the right cerebellar hemisphere on a CT scan on one sheep. A cyst was found in the left frontal lobe on a CT scan on one calf. Microscopically, C. cerebralis cysts were surrounded by a fibrous or epithelial wall that presented necrosis on cerebral sections of both the sheep and the cattle. The CO1-PCR assay yielded a 446 bp band, which was sequenced and phylogenetically analyzed: the results confirmed the presence of T. multiceps. This study reports the first use of CT imaging on naturally infected calves and sheep for diagnosing coenurosis.

Resumo Os objetivos deste estudo foram diagnosticar cenurose por tomografia computadorizada (CT) por imagem de digitalização e caracterização molecular do gene CO1, usando a Reação em Cadeia da Polimerase (PCR). Ovelhas e bezerros foram necropsiados, e uma tomografia computadorizada da região cefálica foi realizada nos animais. Em seguida, cortes microscópicos de cérebro infectado com parasitas foram corados com hematoxilina e eosina e posterior avaliação ao microscópio de luz. Em seguida, o material recolhido de cada cisto cerebral foi fixado em etanol a 70%. A amplificação pela PCR foi realizada utilizando-se o gene mitocondrial CO1. Um total de 60 bezerros e 80 ovelhas foram clinicamente examinados e, desses, 15 bezerros e 38 ovelhas apresentaram sinais de depressão, com movimentos circulares em sentido anti-horário, e desvio da cabeça. Quatro carneiros e uma vitela foram necropsiados, e cistos de C. cerebralis foram detectados nos animais. Um cisto hipodenso foi monitorado no hemisfério cerebelar direito por imagem do CT de um carneiro. O cisto foi encontrado no lobo frontal esquerdo por imagem do CT de um bezerro. Microscopicamente, cistos de C. cerebralis foram envolvidos por uma parede fibrosa ou epitelial, apresentando necrose em ambos os cortes cerebrais de ovinos e de bovinos. O ensaio CO1-PCR produziu uma banda de 446 pb, sequenciado e submetido à filogenia, confirmou ser T. multiceps. Este estudo relata a primeira utilização de imagens de CT em bezerros e ovelhas naturalmente infectados para o diagnóstico de coenurosis.

Clinical, pathological and molecular evaluations and CT scan screening of coenurosis (Coenurus cerebralis) in sheep and calves.

The aims of this study were to diagnose coenurosis by means of computerized tomography (CT) scan imaging and molecular characterization of the CO1 gene using the polymerase chain reaction (PCR). Sheep and calves were necropsied, and CT scans on the cephalic region were performed on the animals. Sections of brain tissue infected with parasites were then stained with hematoxylin and eosin for microscopic examination. Material collected from brain cysts was fixed in 70% ethanol. PCR amplification was carried out using the CO1 mitochondrial gene. A total of 60 calves and 80 sheep were examined clinically and, of these, 15 calves and 38 sheep showed signs of depression, with counterclockwise circling movements and altered head carriage. Four sheep and one calf were necropsied, and C. cerebralis cysts were detected in all of them. A hypodense cyst was monitored in the right cerebellar hemisphere on a CT scan on one sheep. A cyst was found in the left frontal lobe on a CT scan on one calf. Microscopically, C. cerebralis cysts were surrounded by a fibrous or epithelial wall that presented necrosis on cerebral sections of both the sheep and the cattle. The CO1-PCR assay yielded a 446 bp band, which was sequenced and phylogenetically analyzed: the results confirmed the presence of T. multiceps. This study reports the first use of CT imaging on naturally infected calves and sheep for diagnosing coenurosis.

Disease behaviours of sows naturally infected with Taenia solium in Tanzania.

Neurocysticercosis (NCC) is a disease caused by the zoonotic parasite Taenia solium lodging in the central nervous system. Both humans and pigs can get NCC. The impact of the disease in pigs has so far been little explored. The aim of this study was to describe the effect of NCC on social and feeding behaviours as well as the pattern of activity as indicators of reduced welfare in naturally infected sows. In total 13 T. solium naturally infected and 15 non-infected control sows were videotaped for 2 consecutive weeks using close circuit television cameras at research facilities at Sokoine University of Agriculture, Morogoro, Tanzania. Videos were analysed at the beginning, in the middle and at the end of the 2 week recording period. For each time point, videos were analysed during feeding, while the enrichment was provided, and by recording every half an hour the sows' behaviours performed over the course of a whole day. Sows with NCC spent significantly less time at the feeding trough, especially during the second half of the feeding period. Infected sows were also more passive e.g. lying and standing still significantly more during a whole day period and showed social isolation compared to non-infected control sows by performing behaviours more distant to their nearest neighbour. Results of this study indicated that NCC changed the behaviour of infected sows. The behavioural changes are indicative of decreased welfare. Efforts to reinforce the animal welfare aspect are needed as this has so far been neglected.

The association between seizures and deposition of collagen in the brain in porcine Taenia solium neurocysticercosis.

Neurocysticercosis caused by infection with Taenia solium is a significant cause of epilepsy and seizures in humans. The aim of this study was to assess the association between seizures and the deposition of collagen in brain tissue in pigs with T. solium neurocysticercosis. In total 78 brain tissue sections from seven pigs were examined histopathologically i.e. two pigs with epileptic seizures and T. solium cysts, four pigs without seizures but with cysts, and one non-infected control pig. Pigs with epileptic seizures had a larger amount of collagen in their brain tissue, showing as large fibrotic scars and moderate amount of collagen deposited around cysts, compared to pigs without seizures and the negative control pig. Our results indicate that collagen is likely to play a considerable part in the pathogenesis of seizures in T. solium neurocysticercosis.

Severe seizures in pigs naturally infected with Taenia solium in Tanzania.

Neurocysticercosis (NCC) caused by Taenia solium is a serious neurological disease. In humans neurological symptoms have been thoroughly studied and documented, however, there is limited information on clinical signs in pigs infected with T. solium cysticerci. Among the scientific community, it is in fact believed that pigs with NCC rarely show neurological signs. The aim of this study was to describe clinical manifestations associated with NCC in pigs and correlate the manifestations to the number and distribution of cysticerci in brains of naturally infected pigs in Tanzania. Sixteen infected and 15 non-infected control pigs were observed for 14 days during daylight hours, and subsequently videotaped for another 14 consecutive days using close circuit television cameras. All occurrences of abnormal behaviour (trembling, twitching, mouth and ear paralysis, ataxia, dribbling, salivating, eye blinking, walking in circles) were recorded. At the end of the recording period, pigs were slaughtered and their brains dissected, cysticerci counted and locations noted. During the recording period, two infected pigs were observed having seizures. Some of the observed autonomic signs during a seizure were chewing motions with foamy salivation and ear stiffening. Motor signs included tonic muscle contractions followed by a sudden diminution in all muscle function leading to collapse of the animal. Stereotypic walking in circles was observed on several occasions. At dissection, both pigs had a high number of brain cysticerci (241 and 247 cysticerci). The two pigs with seizures were also older (36 months) compared to the others (18.3 months, ± 8.2 standard deviation). Results of this study have shown that pigs with NCC can develop clinical signs and suffer from seizures like humans with symptomatic NCC. Results of this study could potentially open up a new experimental pathway to explore the aetiology of neurological symptoms in humans with NCC associated epilepsy.

Neurocysticercosis in a Rhesus Macaque (Macaca mulatta).

An 8-y-old, intact, male rhesus macaque (Macaca mulatta) was sedated to undergo MRI in preparation for the implantation of cranial hardware. During imaging, 9 focal lesions were noted in the brain and musculature of the head. The lesions were hyperechoic with hypoechoic rims. The animal was deemed inappropriate for neuroscience research, and euthanasia was elected. Gross examination revealed multiple round, thick-walled, fluid-filled cysts (diameter, approximately 0.5 cm) in multiple tissues: one each in the left caudal lung lobe, left masseter muscle, and the dura overlying the brain and 8 throughout the gray and white matter of the brain parenchyma. Formalin-fixed sections of cyst-containing brain were stained with hematoxylin and eosin. Microscopic examination and molecular analysis of the COX1 (COI) gene recognized the causative organism as Taenia solium at 99.04% identity.

Monitoring the outcomes of interventions against Taenia solium: options and suggestions.

There is an increasing interest in reducing the incidence of human neurocysticercosis, caused by infection with the larval stage of Taenia solium. Several intervention trials are currently assessing various options for control of T. solium transmission. A critical aspect of these trials will be the evaluation of whether the interventions have been successful. However, there is no consensus about the most appropriate or valuable methods that should be used. Here, we undertake a critical assessment of the diagnostic tests which are currently available for human T. solium taeniasis and human and porcine cysticercosis, as well as their suitability for evaluation of intervention trial outcomes. Suggestions are made about which of the measures that are available for evaluation of T. solium interventions would be most suitable, and which methodologies are the most appropriate given currently available technologies. Suggestions are also made in relation to the most urgent research needs in order to address deficiencies in current diagnostic methods.

Inflammation Caused by Praziquantel Treatment Depends on the Location of the Taenia solium Cysticercus in Porcine Neurocysticercosis.

BACKGROUND: Neurocysticercosis (NCC), infection of the central nervous system by Taenia solium cysticerci, is a pleomorphic disease. Inflammation around cysticerci is the major cause of disease but is variably present. One factor modulating the inflammatory responses may be the location and characteristics of the brain tissue adjacent to cysticerci. We analyzed and compared the inflammatory responses to cysticerci located in the parenchyma to those in the meninges or cysticerci partially in contact with both the parenchyma and the meninges (corticomeningeal). METHODOLOGY/PRINCIPAL FINDINGS: Histological specimens of brain cysticerci (n = 196) from 11 pigs naturally infected with Taenia solium cysticerci were used. Four pigs were sacrificed after 2 days and four after 5 days of a single dose of praziquantel; 3 pigs did not receive treatment. All pigs were intravenously injected with Evans Blue to assess disruption of the blood-brain barrier. The degree of inflammation was estimated by use of a histological score (ISC) based on the extent of the inflammation in the pericystic areas as assessed in an image composed of several photomicrographs taken at 40X amplification. Parenchymal cysticerci provoked a significantly greater level of pericystic inflammation (higher ISC) after antiparasitic treatment compared to meningeal and corticomeningeal cysticerci. ISC of meningeal cysticerci was not significantly affected by treatment. In corticomeningeal cysticerci, the increase in ISC score was correlated to the extent of the cysticercus adjacent to the brain parenchyma. Disruption of the blood-brain barrier was associated with treatment only in parenchymal tissue. SIGNIFICANCE: Inflammatory response to cysticerci located in the meninges was significantly decreased compared to parenchymal cysticerci. The suboptimal inflammatory response to cysticidal drugs may be the reason subarachnoid NCC is generally refractory to treatment compared to parenchymal NCC.