L.J.S. Harrison

Limitations of current diagnostic procedures for the diagnosis of Taenia solium cysticercosis in rural pigs

The aim of the present study was to evaluate diagnostic procedures for porcine cysticercosis. Sera were obtained from 32 pigs reared in commercial farms, 47 pigs before and after experimental infection, 42 carefully necropsied rural pigs and 191 slaughtered pigs from rural communities in which the presence of the Taenia solium metacestode was assessed by tongue dissection. Sera were analyzed by ELISA to detect antibodies against T. solium antigens and to detect parasite antigens. Most sera from the necropsied rural pigs were also evaluated by the Western blot method. Antigen and antibody ELISA detection assays showed high sensitivity and specificity when applied to sera from pigs reared in commercial farms. In contrast, all methods (Ag-ELISA, Ab-ELISA assays, EITB and tongue inspection) showed lower sensitivity and specificity when applied to the generally lightly infected rurally reared pigs. The probability distribution of cysts in carcasses were also determined. These results emphasize the difficulties in detecting cysticercosis in rural pigs with low levels of cyst burdens.

Prevalence of Taenia solium cysticercosis in swine from a community-based study in 21 villages of the Eastern Cape Province, South Africa

The pork tapeworm, Taenia solium, causative organism of porcine cysticercosis and human neurocysticercosis is known to occur in areas of South Africa including Eastern Cape Province but, despite increasing reports of its occurrence throughout the subregion, the prevalence is yet to be clearly established. The parasite presents a potentially serious agricultural problem and public health risk in endemic areas. The human populations considered to be at highest risk of infection with this zoonotic helminth are people living in rural areas most of whom earn their livelihood wholly or partially through livestock rearing. Here we report on initial results of a community-based study of pigs owned by resource-poor, emerging pig producers from 21 villages in the Eastern Cape Province. Lingual examination (tongue palpation) in live pigs, two enzyme-linked immunosorbent assays (ELISAs), which detect parasite antigen (B158/B60 Ag-ELISA and HP10 Ag-ELISA) and an enzyme immunotransfer blot (EITB) assay, which detects antiparasite antibody, were used to verify endemicity and estimate apparent prevalence. In the absence of a gold standard true prevalence was obtained, using a Bayesian approach, with a model that uses both available data and prior information. Results indicate that the parasite is indeed present in the study villages and that true prevalence was 64.6%. The apparent prevalences as measured by each of the four tests were: 11.9% for lingual examination, 54.8% for B158/B60 Ag-ELISA, 40.6% for HP10 Ag-ELISA and 33.3% for EITB. This base-line knowledge of the prevalence of T. solium in pigs provides information essential to the design and monitoring of sustainable and appropriate interventions for cysticercosis prevention and control.

Diagnosis of porcine cysticercosis: a comparative study of serological tests for detection of circulating antibody and viable parasites

Epidemiological studies of porcine cysticercosis require identification of pigs harbouring viable Taenia solium cysticerci and estimates of the degree of exposure to the parasite in the pig population destined for human consumption. Identification of infected pigs with viable larvae is achieved through detection of their secretory products. However, detectable levels of circulating antibody may also be present in the absence of viable larvae. In this study, both types of tests have been evaluated in groups of pigs experimentally infected with T. solium. Detection of viable cysticerci was achieved using a monoclonal antibody-based (HP10) antigen capture assay. HP10 epitope-bearing antigens have now been demonstrated in T. solium and T. crassiceps cyst fluid and excretion/secretions. Serum antibodies were measured in ELISA assays using two parasite preparations as antigens; T. solium cyst fluid and T. crassiceps cyst fluid antigens bearing the HP10 epitope. Low-background values were obtained with sera from non-infected animals in all the assays used. In heavily infected pigs, both antigens and antibodies were detected at least 29 days and up to 200 days post-infection (pi), while in lightly infected pigs antigen and antibodies were first observed between 61-97 days pi. Thus, the levels of the serum antigen and antibody varied with the intensity of the infection.

Detection of secreted cysticercal antigen: a useful tool in the diagnosis of inflammatory neurocysticercosis

Neurocysticercosis is a common parasitic disease of the human central nervous system. It is particularly prevalent in developing countries, where it has a serious public health and economic impact. A major diagnostic problem with neurocysticercosis is its pleomorphic nature. Conventional diagnosis of neurocysticercosis still requires brain-computed tomography and/or magnetic resonance imaging, which are definitive but often prohibitively expensive and inaccessible in endemic areas. Herein, the monoclonal antibody HP10 antigen-trapping enzyme-linked immunosorbent assay, which has been used successfully to detect viable Taenia solium cysticercosis, was evaluated using cerebrospinal fluid (CSF) from Mexican neurocysticercosis patients with various defined pathologies. Sensitivity was higher in cases of inflammatory compared with non-inflammatory disease (94.1% vs. 33.3%) and in cases of multiple- compared with single-cyst cysticercosis (85% vs. 33.3%). Positivity was a strong indicator of active, inflammatory, multiple-cyst neurocysticercosis detecting 100% (15/15) of such cases. The overall specificity, as determined using CSF samples from patients with other neurological symptoms, was 97.7% (42/43). Since the assay only detects viable infection, it is of known value in the follow-up of treated patients to determine whether treatment has been successful. Thus, antigen detection may be of particular value in the assessment of symptomatic patients, who may potentially benefit from rapid treatment.

Diagnosis of Taenia saginata cysticercosis in Kenyan cattle by antibody and antigen ELISA

Sera from calves, either experimentally or naturally infected with Taenia saginata, were screened for an antibody response to T. saginata, and for parasite antigen, by enzyme-linked immunosorbent assays (ELISAs). An antibody response was detected by 3 weeks post infection (p.i.), rose to a peak at 10-12 weeks p.i., and was still in evidence 1 year p.i. Parasite antigen was first detected 4-7 weeks p.i. and persisted until the end of the experiment, over 1 year p.i. In the experimentally infected animals, cattle with 14 or more live cysticerci had detectable levels of parasite antigen in their sera at slaughter, while animals with live cyst burdens ranging from 0 to 4 were negative. Furthermore, levels of circulating antigen were positively correlated with live cysticercus burden in the experimental animals. In naturally infected cattle, 83% (5/6) of those with 30 or more live cysts, and 22% (5/23) of those with 1-29 live cysts, could be detected by the ELISA for parasite antigen, although no significant correlation between antigen level and live cyst burden could be detected. Antibody levels were not found to be associated with cyst burdens in either experimentally or naturally infected cattle. In slaughterhouse cattle, the antigen assay was almost three times as sensitive as meat inspection. However, there was no agreement between cattle found positive at meat inspection and those found positive by the antigen detection ELISA. One possible reason is that the ELISA only detects live cysts, while lesions left by dead cysts are more noticeable at meat inspection. The mouse monoclonal antibody-based antigen detection ELISA is of value for the diagnosis of naturally occurring, viable, T. saginata cysticercosis in live cattle and has an immediate application for field based epidemiological studies designed to determine prevalence.

Ag-ELISA and PCR for monitoring the vaccination of cattle against Taenia saginata cysticercosis using an oncospheral adhesion protein (HP6) with surface and secreted localization.

ATaenia saginata oncosphere-derived adhesion protein (HP6) with surface and secreted localization was used to successfully vaccinate calves against oral challenge withT. saginata eggs. In contrast, vaccination using a combination ofT. saginata oncosphere-derived peptides, selected on the basis of their antigenic index, and including three derived from the HP6 molecule (HP6-1, HP6-2 and HP6-3), was unsuccessful. This either indicated that the wrong peptides were selected or, in the case of the HP6 protein, that the protective epitope is conformational in nature. The protection experiments were monitored using a parasite antigen detection ELISA (HP10 Ag-ELISA), which allowed the early determination of the success of the vaccination protocol, subsequently confirmed at autopsy. PCR assays were used for the first time to confirm the presence ofT. saginata DNA in lesions recovered at autopsy and thus verify the parasite origin of the lesions.

Taenia saginata derived synthetic peptides with potential for the diagnosis of bovine cysticercosis

Immunity in Taeniids is predominantly antibody mediated and thus many serological immuno-determinants will have potential in both protection and diagnosis. The antigenicity of six peptides derived from four potentially protective molecules cloned from a Taenia saginata oncospheres cDNA library have been evaluated as targets for the specific diagnosis of bovine cysticercosis. The six peptides consist of: two peptides (HP6-2 and HP6-3) derived from the sequence of the 18 kDa surface/secreted oncospheral adhesion antigen identified by McAb-HP6, two peptides (Ts45W-1 and Ts45W-5) derived from the sequence of the T. saginata homologue of the T. ovis 45W protective gene family, one peptide (TS45S-10) derived from a T. saginata sequence with significant similarity to the T. ovis 45S protective antigen, and one peptide (TEG-1) derived from the sequence of the T. saginata homologue of Echinococcus spp. main surface protein. Longitudinal studies indicate that T. saginata infected cattle respond to all six peptides by 3-4 weeks post-infection and that the antibody levels remain high for at least 12 weeks post-infection. As protection against Taeniid parasites is predominantly antibody mediated, some of these six peptides may be of value as immuno-prophylactic tools and hence also in assays to determine resistance to infection with the parasite. For diagnosis, on the other hand, only three peptides (HP6-2, TEG-1 and Ts45S-10) performed with the necessary sensitivity and specificity to determine exposure to infection with T. saginata, and now merit an exhaustive evaluation prior to employment as routine diagnostic tools.

Control of fasciolosis in stall-fed buffaloes by managing the feeding of rice straw.

Fasciolosis, caused by Fasciola gigantica, presents major disease and production problems, particularly in the rice producing areas of South and South-East Asia. In Nepal, buffaloes are commonly stall-fed and the most usual fodder is rice straw, which is produced in large quantities in the area. Here we describe a relatively simple way of controlling fasciolosis in these animals. We demonstrate that infectious metacercariae of F. gigantica are concentrated in the bottom part of the rice straw and that infection can be controlled by cutting the rice straw in half, and then feeding the parasite free top half of the straw first. The bottom part of the rice straw should ideally be stored for at least 4 months after harvest to allow the parasites to die before it is used as fodder.

Risk factors associated with taeniosis-cysticercosis in rural farming communities in Gauteng Province, South Africa

The aim of this study was to determine which of the livestock management and human practices known to be risk factors associated with taeniosis-cysticercosis occur in Gauteng Province. A questionnaire survey was conducted in two regions of Gauteng Province, Germiston and Pretoria. Results revealed that almost 20% of the interviewed farmers do not have toilets, most of them let their animals roam freely during the day for grazing and scavenging, and 47% use streams as the water source for their animals. This may create an infection opportunity through ingestion of Taenia-contaminated herbage or water. Furthermore, 26% mentioned that their animals might have access to human excreta. More than 70% of farmers in the province slaughter cattle and pigs for their own consumption without inspecting meat for cysticercosis. Only a few of the interviewed farmers in both regions were aware of the taeniosis-cysticercosis complex. Backyard slaughtering, consumption of uninspected meat by the public, poor livestock management, and limited sanitation in rural communities of Gauteng Province are identified as risk factors associated with the occurrence of Taenia saginata and Taenia solium infections in the province. Taenia saginata and T. solium are considered to have a global distribution; therefore, these risk factors may be applicable globally, not just in Gauteng Province. Programs on public awareness with regard to transmission and prevention of Taenia infections as well as more detailed studies on risk factors of taeniosis-cysticercosis are recommended.