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.

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.

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.

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.

Distribution and histopathological changes induced by cysts of Taenia solium in the brain of pigs from Tanzania.

Neurocysticercosis (NCC) caused by Taenia solium cysts is a frequent but neglected parasitic disease of the central nervous system (CNS) worldwide. The aim of this study was to describe anatomical locations of cysts in the CNS and the corresponding inflammation. A total of 17 naturally infected pigs were used to evaluate the distribution of cysts and, of these, seven were used to evaluate the corresponding inflammation further, through histopathology. Clinical signs in the pigs included dullness, sluggishness, somnolence, apathy and loss of consciousness. Cysts were distributed in all cerebral lobes, i.e. 39.7% in the frontal lobe, 20.3% in the parietal lobe, 20.0% in the occipital lobe and 19.7% in the temporal lobe, and only 0.4% in the cerebellum. No cysts were found in the spinal cord. Cysts were localized as follows: 47.9% in the dorsal subarachnoid, 46.9% in the parenchyma, 4.4% in the subarachnoid base and 0.9% in the ventricles. The results of the histopathology revealed lesions in an early inflammatory stage, i.e. stage I, in all anatomical locations except for two, which showed more of an inflammatory reaction, stage III, in one pig. It was concluded that clinical signs in pigs were neither pathognomonic nor consistent. These signs, therefore, cannot be used as a reliable indicator of porcine NCC. Furthermore, T. solium cysts were found to be in abundance in all cerebral lobes, and only a few were found in the cerebellum. Regarding the inflammatory response, no significant differences were found in the location and total number of cysts. Thus, further studies are needed to explain the determinants of cyst distribution in the CNS and assess in detail clinical signs associated with porcine NCC.

Control of Taenia solium taeniasis/cysticercosis: past practices and new possibilities.

Neurocysticercosis continues to be a major health burden on humans living in many regions of the world, despite the availability of highly effective taeniacides and identification of the cause, Taenia solium, as being potentially eradicable. Several T. solium control trials have been undertaken, generally achieving limited success and none that has been fully documented has achieved what was demonstrated to be a sustainable level of disease control. Pigs act as intermediate hosts for T. solium and two new control tools have become available for application in pigs - single-dose oxfendazole treatment of porcine cysticercosis and the TSOL18 vaccine. Three potential intervention scenarios for pigs are compared for control of cysticercosis, using either oxfendazole or vaccination. A control scenario involving vaccination plus oxfendazole treatment delivered at 4 monthly intervals was predicted to achieve the best outcome, with no pigs slaughtered at 12 months of age having viable T. solium cysticerci. Now that new control tools are available, there are opportunities to concentrate research attention on evaluation of novel control scenarios leading to the implementation of effective and sustainable control programmes and a reduction in the global burden of neurocysticercosis.

Disruption of the blood-brain barrier in pigs naturally infected with Taenia solium, untreated and after anthelmintic treatment.

Neurocysticercosis is a widely prevalent disease in the tropics that causes seizures and a variety into of neurological symptoms in most of the world. Experimental models are limited and do not allow assessment of the degree of inflammation around brain cysts. The vital dye Evans Blue (EB) was injected to 11 pigs naturally infected with Taenia solium cysts to visually identify the extent of disruption of the blood-brain barrier. A total of 369 cysts were recovered from the 11 brains and classified according to the staining of their capsules as blue or unstained. The proportion of cysts with blue capsules was significantly higher in brains from pigs that had received anthelmintic treatment 48 and 120h before the EB infusion, indicating a greater compromise of the blood-brain barrier due to treatment. The model could be useful for understanding the pathology of treatment-induced inflammation in neurocysticercosis.

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.

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.

The immune response in Taenia solium neurocysticercosis in pigs is associated with astrogliosis, axonal degeneration and altered blood-brain barrier permeability.

Immunohistochemistry was used to examine the immuno-pathological changes and the extent of neuronal damage caused by either viable or dead Taenia solium cysticerci during porcine neurocysticercosis. Thirty pig brains with cerebral cysticercosis and 5 brains from T. solium free pigs were used in this study. Results revealed extensive astrogliosis, neuronal and mostly axonal damage in both early (grade I) and late (grades III and V) lesions as evidenced by an increased expression of glial fibrillary acidic protein (GFAP) and neurofilament protein (NFP). In many late lesions, astrocyte end-feet formed glial scars that surrounded the dead parasite. Rapid angiogenesis resulted in blood vessels lacking astrocyte end-feet suggesting loss of blood-brain barrier (BBB) hence allowing an influx of peripheral blood immune cells such as eosinophils, macrophages, CD3+ T cells, B lymphocytes and plasma cells into lesions. This study showed that porcine NCC was associated with severe nervous tissue damage, the host response of which is a collaborative effort between the local and peripheral immune responses comparable to that observed in human NCC. Results further implied that porcine NCC could be a useful model for understanding the course of NCC in human as well as provide useful information for therapeutic and/or immune strategies.

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.

Human and porcine neurocysticercosis: differences in the distribution and developmental stages of cysticerci.

OBJECTIVE: To describe and compare the clinical impacts of neurocysticercosis (NC) caused by Taenia solium in humans and pigs. METHODS: Comparative study of the brains of 16 asymptomatic pigs and 35 human NC cases (15 asymptomatic and 20 symptomatic). RESULTS: In humans, cysticerci were more frequently located in the ventricles and subarachnoid space at the base of the brain (11.8%vs. 1.6%; P = 0.001 and 25.9%vs. 0%; P < 0.0001, respectively) while in pigs, cysticerci were more frequently found in the parenchyma (44.4%vs. 7.6%; P < 0.0001). In human brains, 75.9% of the cysticerci were calcified, while in pigs all cysticerci were in the vesicular stage. CONCLUSION: The duration of infection and the host-parasite relationship (such as immune reactivity and brain haemodynamics) differ between humans and pigs. This may account for the different distribution and stage of the cysticerci among humans and pigs.

Control of neurocysticercosis by routine medical and veterinary services.

Taenia solium neurocysticercosis and ocular cysticercosis are two of the most devastating parasitic infections, which need to be controlled for medical and economic reasons. This paper discusses why control measures are not implemented adequately in endemic areas and proposes simple operational interventions, based on focus-oriented chemotherapy of T. solium taeniasis using existing health care infrastructure and improved collaboration between medical and veterinary services. These interventions can be quickly and easily implemented (irrespective of other applicable control measures), with medical and veterinary staff being adequately trained, and safe, effective and cheap taenicides made available.

Neurocysticercosis: a possible cause of epileptiform seizures in people residing in villages served by the Bethanie clinic in the North West Province of South Africa.

A study to detect human taeniasis and cysticercosis was conducted in 4 village communities served by the Bethanie clinic in the North West Province, based on reports of people being diagnosed there with epileptiform episodes. Many home owners in the villages rear pigs in small numbers for both meat availability and an immediate income from live pig or pig meat sales. The primary aim of the work was to conduct in the study area a census of all small scale pig producers and a survey of rural village consumers, both by means of a structured questionnaire. The former reviewed pig husbandry practices, slaughter and marketing of pigs and the latter provided information on pork consumption, sanitation as well as people's basic knowledge of Taenia solium. Stool samples from consenting participants were screened by a contracted approved laboratory for IR solium. A descriptive analysis of retrospective data was conducted at the Bethanie clinic to determine the proportional morbidity of neurocysticercosis from the medical records of patients diagnosed with seizures in an attempt to establish possible sources of infection and routes of transmission. In addition, the total pig population in the study area was determined more accurately and the prevalence of cysticercosis investigated in pigs subjected to meat inspection at an approved abattoir. The questionnaires revealed a poor understanding of the disease, poor sanitation and hygiene, poor methods of pig husbandry and poor meat inspection and control in rural smallholder communities. There was no significant statistical difference in the proportion of households reporting evidence of epilepsy and owning pigs and those that did not. There is a strong evidence of a tendency towards an association between epilepsy, consumption habits and some identified epidemiological risk factors.

Two rare clinical manifestations of coenurosis in sheep.

Two rare clinical manifestations of coenurosis in sheep are reported. (i) A case of partial seizure disorder in a ram of 11 months old. During seizure episodes the animal lay down in lateral recumbency displaying initially a stuporous condition and subsequently began to revolve its head from the base of the cervix. At the necropsy of the case, Coenurus cerebralis cyst (young bladder worm) was found dorsally inside the brainstem, in the site of the tectum mesencephaly. (ii) The second-reported manifestation was a bacterial meningoengephalitis that was witnessed in two lambs of 6-7 weeks old. The lambs displayed lateral recumbency with seizure activity. At necropsy, meningoencephalitis with congestion and abscesses were observed in both of them. Interestingly, C. cerebralis cysts were also found in both brains. Streptococcus dysgalactiae was isolated from the abscesses. Possibly, S. dysgalactiae translocation of the blood-brain barrier was facilitated by the migration of the immature stages of C. cerebralis to and through the brain.

A survey of Taenia multiceps coenurosis in Sardinian sheep.

A survey was carried out to assess the occurrence of Coenurus cerebralis infection in Sardinian sheep. A prevalence of 0.35% was observed when 566 regularly slaughtered sheep were examined. However, in 120 sheep with suspected symptoms of coenurosis examined from November 2001 to October 2002, a total of 299 cerebral coenurosis lesions were observed with an incidence of 1% per year. Lesions were classified as migratory, cystic and secondary. Most migratory lesions were found in sheep aged 3-6 months. Cavitary lesions containing cysts in different developing stages were found with high incidence per year in sheep aged 7-12 months. Secondary lesions due to the development of Coenurus were most frequent in sheep aged 19-36 months. Most sheep were found infected in spring and in early summer, between March and June. Most lesions were located in the cortex. The mean number of protoscolices per cyst was 149 (range 10-370).

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.

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.