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.

Sero-prevalence of Taenia spp. infections in cattle and pigs in rural farming communities in Free State and Gauteng provinces, South Africa.

The aim of this study was to determine sero-prevalence of bovine and porcine cysticercosis in cattle and pigs in rural farming communities in Free State and Gauteng Provinces, Republic of South Africa. Blood samples were collected for a period of twelve months from live cattle (n=1315; 1159) and pigs (n=436; 240) and the serum extracted and stored before analysis by a monoclonal antibody based (HP10) antigen detection ELISA. Results revealed a generally high sero-prevalence and wide distribution throughout the two provinces with Free State having a higher sero-prevalence in both cattle and pigs (23% and 34%) than Gauteng province (15% and 14%). Consumption of infected meat that is either not inspected/missed at meat inspection; poor livestock management practices and limited sanitation in rural communities might have contributed to the occurrence of Taenia spp. infections in the two provinces. It is therefore, recommended that cysticercosis status of animals be established before slaughter. This would assist in ensuring that infected animals are not slaughtered for human consumption or zoonosis preventive measures are taken. Furthermore, public awareness programs on life cycles of T. saginata, T. solium and T. hydatigena and the use of more sensitive diagnostic tools are recommended as part of effective control strategies against taeniid infections.

Histological and molecular biology diagnosis of neurocysticercosis in a patient without history of travel to endemic areas: case report.

BACKGROUND: in endemic areas, neurocysticercosis appears mainly as a single, large, spherical and non-enhancing intracranial cyst. CASE PRESENTATION: an atypical case of neurocysticercosis (NCC) in a French Caucasian, without history of travel to endemic areas, was confirmed by histology and molecular speciation. Imaging was atypical, showing several hook-bearing scolices visible in the cyst, while the serology employed was non-contributary. CONCLUSIONS: NCC should be considered when multiple taeniid scolices are observed within the same cystic lesion.

Neurocysticercosis: detection of Taenia solium DNA in human cerebrospinal fluid using a semi-nested PCR based on HDP2.

Human neurocysticercosis (NC) is caused by Taenia solium larvae lodged in the central nervous system. This disease is usually diagnosed by radiology but the results are not always clear-cut and so immunological assays are often also used. A semi-nested PCR, based on the non-coding HDP2 sequence of T. saginata, has now been developed for detecting DNA from T. solium cysticerci and confirming NC. This PCR, which amplifies a 171-bp T. solium product, allowed the specific detection of just 174 attograms of T. solium DNA. The efficacy of the PCR was tested using cerebrospinal fluid (CSF) from neurological patients, including 46 confirmed Mexican cases of NC and 32 patients from non-endemic Spain. Eighteen of the confirmed cases [including 10 (71%) of the 14 with vesicular extraparenchymal cysticerci and four (17%) of the 24 with damaged cysticerci] and two (33%) of the six patients with 'uncertain' diagnosis (in whom a diagnosis of NC could not be established by radiological and immunological studies) were found PCR-positive. The 36 patients known to have neurological problems other than NC were found PCR-negative. The HDP2 PCR offers a new tool in the diagnosis of NC and in exploring the pathogenesis of this serious disease.

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.

Molecular identification of Echinococcus granulosus genotypes (G1 and G7) isolated from pigs in Mexico.

With the aim of genotyping Echinococcus granulosus cysts found in Mexican livestock, we collected hydatid cysts from the livers and lungs of pigs in slaughterhouses in the state of Morelos, Central Region of Mexico. DNA was extracted from the parasites and examined by polymerase chain reaction (PCR) of rDNA internal transcribed spacer 1 (ITS1-PCR), Eg9-PCR, Eg16-PCR, and PCR-restriction fragment length polymorphism (PCR-RFLP). In addition, fragments of the genes coding for mitochondrial cytochrome c oxidase subunit 1 (CO1) and NADH dehydrogenase 1 (ND1) were sequenced. Two different genotypes of E. granulosus were unequivocally identified, the common sheep genotype, G1, and the common pig genotype, G7. The G1 genotype of E. granulosus has not been previously demonstrated in Mexico. Because of its recognized infectivity in humans, G1 genotype is a direct threat to human health and its presence in Mexico is consequently of immediate public health importance and epidemiological relevance.

Detection of HP10 antigen in serum for diagnosis and follow-up of subarachnoidal and intraventricular human neurocysticercosis.

INTRODUCTION: Neurocysticercosis (NC), a parasitic disease caused by Taenia solium, may be either asymptomatic or show a mild to severe clinical picture with intracranial hypertension. The most severe form of the disease is caused when viable cysticerci are localised in the ventricles or in subarachnoidal cisterns at the base of the skull. Detection of the secreted metacestode antigen HP10 in cerebrospinal fluid is a sensitive and specific method for the diagnosis of these severe NC cases. OBJECTIVE AND METHODS: To evaluate the validity of HP10 antigen detection ELISA when applied to serum, using paired serum and cerebrospinal fluid samples from 116 radiologically and clinically characterised NC patients. RESULTS: The HP10 antigen assay exhibited a similarly high sensitivity in identifying severe NC cases from sera (84.8%) and CSF (91.3%). In contrast, HP10 antigen was rarely detected in asymptomatic or mild NC cases (3 of 57). Importantly, the HP10 antigen assay applied to serum showed high specificity (94%) when used in 126 serum samples of non-NC subjects from an endemic community with a confirmed coproparasitological diagnosis of intestinal parasitic infections. Finally, the HP10 assay also proved to be of value in the follow-up of treated patients. CONCLUSION: This study confirms that detection of the metacestode HP10 antigen in serum is a useful tool for diagnosis and follow-up of patients with severe forms of NC treated with cysticidal drugs.

Differential molecular identification of Taeniid spp. and Sarcocystis spp. cysts isolated from infected pigs and cattle.

In the present work, the species-specific identification of Taeniid spp. cysticerci and sarcocystis cysts isolated from infected pigs and cattle was achieved by PCR. In particular: (i) multiplex-PCR derived from HDP2 DNA fragment, specific for Taenia saginata/Taenia solium; (ii) PCRs and PCR-RFLPs of the rDNA internal transcribed spacers 1 and 2 (ITS1 and ITS2) for the differential diagnosis of taeniids; (iii) PCR derived from the 18S rRNA gene and sequencing, specific for Sarcoystis spp. The combined application of these three PCR protocols provided an unequivocally specific diagnosis of T. saginata, T. solium, T. hydatigena, Sarcocystis hominis and Sarcocystis suihominis, and may have practical application in the identification of calcified degenerating or morphologically dubious cysts, for example in the slaughter house situation or in human biopsy samples.

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.

A Taenia saginata oncosphere-derived adhesion protein (HP6) with surface and secreted localization was used to successfully vaccinate calves against oral challenge with T. saginata eggs. In contrast, vaccination using a combination of T. 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 of T. saginata DNA in lesions recovered at autopsy and thus verify the parasite origin of the lesions.

Control of Taenia saginata by post-mortem examination of carcasses.

BACKGROUND: A study to curb transmission cycle of a zoonotic Taenia cestodiasis between humans and cattle is presented. OBJECTIVE: To evaluate the reliability of meat inspection procedure in detecting carcasses of cattle with T. saginata cysticercosis. METHODS: A total of 55 cattle divided into two groups of artificially (n=30) and naturally (n= 25) infested animals were utilized. Total dissection method was used as a gold standard of validity. RESULTS: Meat inspection insensitively revealed cysticerci in 12 carcasses in each group compared with 24 and 23 carcasses revealed by total dissection in natural and artificial infestations, respectively. Sites of oncosphere invasion showed great variations with the two groups of cattle. In the predilection sites, most cysticerci were found in the heart, Triceps brachii, tongue and head muscles in that order. However, non-predilection sites (neck and back, hind limbs, chest, pelvic and lumbar regions, lungs and liver) considerably harboured high numbers of cysticerci. Observations indicated that except for the dead, degenerate or calcified cysticerci a careless meat inspector will most likely miss out quite a number of viable cysticerci, which blend the pinkish-red colour of the meat and be passed on for human consumption, becoming the source of bovine cysticercosis. CONCLUSIONS: The results confirmed that in spite of the time and efforts taken by meat inspectors looking for cysticerci at specified predilection sites of carcasses, this method is insensitive and inaccurate. To effectively improve meat inspection procedures, there is need to increase the area and number of predilection sites observed during inspection and vary them according to the nature of the animals, their husbandry history and the target human population for consumption. In addition, other control approaches such as vaccination, chemotherapy and immunodiagnosis should be developed and implemented to complement meat inspection procedures.

Taenia solium cDNA sequence encoding a putative immunodiagnostic antigen for human cysticercosis.

A T. solium metacestode cDNA library was prepared and antibody screened to obtain recombinant antigens, which could be used for the neurocysticercosis diagnosis. The F18 clone was selected and sequenced, and the full length cDNA characterised as well as the genomic structure from the gene. F18 is a single copy gene that spans approximately 6.1 kb and contains five exons and four introns. The F18 cDNA has a 690-nucleotide open reading frame that encodes a putative polypeptide of 229 amino acids with a predicted molecular mass of 26.06 x 10(3) M(r). The F18 recombinant protein was obtained and purified by affinity chromatography using pGEX system (G-F18) and pQE system (H-F18). The purified G-F18 fusion protein showed the best results when it was used in ELISA with sera from neurocysticercosis patients.

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.

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.

Circulating parasite antigen in patients with hydrocephalus secondary to neurocysticercosis.

End stages of neurocysticercosis include residual intraparenchymal brain calcifications and hydrocephalus. Although brain calcifications alone have a benign prognosis, hydrocephalus is frequently associated with chronic inflammation and intracranial hypertension, together with a protracted clinical evolution, and may lead to patient deaths. By using a monoclonal-based antigen detection enzyme-linked immunosorbent assay, we measured the levels of circulating parasite antigen in the sera of 56 patients with neurocysticercosis: 27 with calcifications only and 29 with hydrocephalus. The assay gave positive results in 14 of 29 patients with hydrocephalus but was consistently negative in patients with calcifications. Circulating parasite antigen in hydrocephalus secondary to neurocysticercosis indicates the presence of live parasites in these patients and thus a potential benefit from antiparasitic therapy.

Serological evidence for recent exposure to Taenia solium in Venezuelan Amerindians.

This study examined the seroprevalence and serum antibody isotype profile for Taenia solium cysticercosis in an Amerindian community in the Amazonas state of Venezuela. An antigen-trapping enzyme-linked immunosorbent assay (Ag-ELISA) was used to detect viable cysticercosis. Indirect ELISA (Ab-ELISA) and enzyme-linked immunoelectrotransfer blot (EITB) was performed by using antigens prepared from T. solium metacestodes to detect anti-parasite antibodies. The Ag-ELISA and Ab-ELISAs revealed 64.7% and 79.0% seropositivity, respectively, in the Amerindian population. Immunoglobulin (Ig) M was the predominant antibody class, suggesting recent infection. In comparison sera from, clinically defined, hospital neurocysticercosis cases revealed only 27% seropositivity by Ag-ELISA, compared with 86-92% seropositivity by Ab-ELISA, and IgG4 was the predominant antibody subclass detected. The EITB antigen recognition patterns of the hospitalized patients were very similar to that of the Amerindians, confirming exposure to the parasite. These results, combined with the predominance of IgM antibody responses and the marked detection of secreted products of viable parasites, strongly suggest that recent exposure to T. solium had occurred in the Amerindian population.

Serodiagnosis of bovine cysticercosis by detecting live Taenia saginata cysts using a monoclonal antibody-based antigen-ELISA.

An ante mortem antigen-ELISA-based diagnosis of Taenia saginata cysticercosis was studied in artificially (n = 24) and naturally (n = 25) infected cattle with the objective of further validating the assay as a field diagnostic test. Based on total dissection as the definitive method of validity, the assay minimally detected 14 live cysticerci in artificially infected calves and 2 in naturally infected steers. In natural infections, the minimum number of live cysticerci consistently detected by Ag-ELISA was 5 while in artificial infections it was above 14. However, other animals with 12 and 17 live cysticerci in artificially infected calves, and 1 and 2 live cysticerci in naturally infected steers, escaped detection for unknown reasons. Animals harbouring dead cysticerci gave negative reactions in the assay as was the case in non-infected experimental control calves. There was a statistically significant positive linear correlation between Ag-ELISA optical density values and burdens of live cysticerci as obtained by total dissection of both artificially infected calves (r = 0.798, n = 24; P < 0.05) and naturally infected steers (r = 0.631, n = 25; P < 0.05). These results clearly show the potential effectiveness of ante mortem monoclonal antibody-based antigen detection ELISA in the diagnosis of bovine cysticercosis in cattle. Its value lies in the diagnosis of infection in cattle as a screening test in a herd, rather than as a diagnostic test at the individual level, due to false positive and negative reactions. In a herd of heavily infected cattle, the assay may, however, provide for individual diagnosis. Nevertheless, more work is recommended to increase its sensitivity so as to be able to diagnose light infections consistently in the field.

Serum antigen detection in the diagnosis, treatment, and follow-up of neurocysticercosis patients.

The efficacy of albendazole (ABZ) treatment for human neurocysticercosis (NCC) was assessed by using a monoclonal antibody-based parasite antigen detection ELISA which specifically detects the products of living cysticerci in human serum. The assay displayed 85% diagnostic sensitivity, detecting 39 of 46 NCC cases. Only patients with a single viable cyst or only enhancing lesions (degenerating parasites) were seronegative. Specificity of the assay was 92% (23/25) when tested in healthy Peruvian volunteers. In 'cured' patients, in whom all parasites died after ABZ therapy, parasite antigen levels fell sharply by 3 months post treatment. This pattern was not observed in patients refractory to treatment. The sensitivity of the assay with serum samples, and its ability to identify successfully treated patients, make this monoclonal antibody-based ELISA the test of choice for the follow-up of NCC cases.

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.

A specific antigen-detection ELISA for the diagnosis of human neurocysticercosis. The Cysticercosis Working Group in Peru.

An enzyme-linked immunosorbent assay (ELISA) for the detection of antigen secreted by viable Taenia solium metacestodes (Ag-ELISA) was applied to 43 pre-treatment and 47 follow-up cerebrospinal fluid (CSF) samples from Peruvian patients with neurocysticercosis demonstrated by computed tomography and enzyme-linked immunoelectrotransfer blot assay. The sensitivity of the assay was 86%. Negative pre-treatment results in the Ag-ELISA test were restricted to patients with only a single live cyst or only enhancing lesions. Patients with hydrocephalus had higher levels of circulating antigen. There was no difference between antigen levels in CSF taken before and immediately after treatment (day 14). Levels of parasite antigen were significantly positively correlated with the number of live cysts detected by tomography and were also proportional to the number and intensity of antibody reactions recognized by the immunoblot diagnostic test. In contrast, there was a negative correlation with the number of enhancing lesions revealed by tomography, supporting the hypothesis that enhancing lesions correspond to a terminal, moribund stage of the parasite. The use of antigen-detection tests specific for viable metacestodes has immediate utility in the clinical context, not only providing important information on the viability of the parasites but also leading to an improved understanding of the pathogenesis of neurocysticercosis before and after drug treatment.

Sequence and preliminary characterisation of a Taenia saginata oncosphere gene homologue of the small heat-shock protein family.

During antibody screening of a Taenia saginata oncosphere cDNA library a clone (R-Tso2) sharing a high degree of homology at both the DNA and amino acid levels with the small heat-shock protein (shsp) family was identified. The R-Tso2 clone was a full-length sequence (1162 bp) with an open reading frame of 945 bp and 314 amino acids, corresponding to a deduced molecular mass of 35.6 kDa and isoelectric point of 5.6. R-Tso2 had the highest degree of homology with the Schistosoma mansoni major egg antigens, showing the characteristic shsp 100 amino-acid sequence motif duplicated. The R-Tso2 expression product was not immuno-precipitated by any serum from a panel of serum samples obtained from bovine, porcine and human hosts suffering from either T. saginata or T. solium cysticercosis.