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.

Relative economic benefits of tactical anthelmintic treatment and urea-molasses block supplementation of Boer goats raised under extensive grazing conditions at Onderstepoort, Pretoria, South Africa.

The potential economic benefits of combining tactical anthelmintic treatment for gastrointestinal nematodes and nutritional supplementation with urea-molasses blocks were examined in Boer goats raised under extensive grazing conditions in the summer rainfall area of South Africa. Eight groups of nine goats were monitored over a 12-month period from 1 October 2002 to 9 October 2003. Ad libitum nutritional supplementation with urea-molasses blocks was provided when the goats were housed at night, during the summer (wet season--December 2002 to February 2003), and/or the winter (dry season--June 2003 to August 2003). All the goats were treated symptomatically for Haemonchus contortus infection when deemed necessary by clinical examination of the conjunctiva for anaemia using the FAMACHA system. Half the groups were tactically treated for gastrointestinal nematodes in mid-summer (28 January 2003). Under the symptomatic treatment, climatic and extensive grazing conditions encountered during the trial, feed supplementation in the winter dry season had the greatest economic benefit and is therefore recommended. Tactical anthelmintic treatment afforded no additional advantage, but the nematode challenge was low.

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.

Benefits of urea-molasses block supplementation and symptomatic and tactical anthelmintic treatments of communally grazed indigenous goats in the Bulwer area, Kwazulu-Natal Province, South Africa.

This study was carried out with the cooperation of farmers owning communally grazed indigenous goats in southwestern KwaZulu-Natal Province, South Africa, where farmers had identified poor reproductive performance in their herds as one of their major problems. The aim was to quantify the effects of 3 interventions and the interaction between these interventions on goat productivity and gastrointestinal nematode infection. The interventions were: urea-molasses block supplementation during the dry winter seasons of 2004 and 2005, tactical anthelmintic treatment with ivermectin (400 microg/kg) during the wet summer period (on 3 January 2005) and symptomatic treatment with ivermectin (400 microg/kg) of all goats judged anaemic throughout the entire study period. The FAMACHA system was used as a gauge of anaemia. It was noted that goats considered anaemic tended to remain so throughout the study period. The tactical anthelmintic treatment was effective as it markedly reduced (P = 0.066) the summer peak in faecal egg counts and is therefore recommended. By contrast, while the urea-molasses block supplementation appeared to reduce the faecal egg counts immediately following the 2004 supplementation (P < 0.05), this did not hold true in 2005. Interestingly, in the tactically treated anaemic goats, the improvement in the number of kids suckled per doe year-on-year tended to be greater than in the non-anaemic goats. It is considered that the routine symptomatic treatment of anaemic goats may have been a key factor. More detailed investigations into the routine symptomatic treatment of anaemic goats are therefore recommended.

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.

The serum glucose and beta-hydroxybutyrate levels in sheep with experimental Fasciola hepatica and Fasciola gigantica infection.

The influence of Fasciola hepatica and Fasciola gigantica infection on serum glucose and beta-hydroxybutyrate (beta-HOB) in sheep was evaluated. This was done by setting up two groups of sheep. The first group (n=13) was split in two sub-groups, one experimentally infected with F. hepatica (n=9) and the other (n=4) as uninfected control. A second group consisting of a sub-group experimentally infected with F. gigantica (n=9) the other sub-group (n=6) left as uninfected control was also set up. The results of weight gain, parasitological and serum liver enzymes activity (glutamate dehydrogenase [GLDH] and gamma glutamyltransferase [gamma-GT]) used in monitoring the infection showed that all infected animals developed fasciolosis. It was observed that a reduction in serum glucose levels was significantly lower (p<0.05) in F. hepatica infected sheep than in uninfected control sheep starting from 5 weeks post-infection (wpi) to the end of the experiment. Similar reduction was recorded in F. gigantica infected sheep between 8 and 19 wpi. In contrast, serum beta-HOB levels were elevated in F. hepatica infected sheep between 6 and 16 wpi and in F. gigantica infected sheep between 7 and 15 wpi. It would appear from these serum glucose and beta-HOB levels that fasciolosis does lead to energy deficiency (low glucose) and ketosis (increased beta-HOB). The decrease in serum glucose and increase in serum beta-HOB levels in infected sheep may help in understanding the interaction between fasciolosis and nutritional status of infected ruminants especially in young growing animals.

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.

Serum antibody isotype responses of Fasciola-infected sheep and cattle to excretory and secretory products of Fasciola species.

This study investigated the immunoglobulin isotype responses of sheep and cattle chronically infected with Fasciola hepatica and Fasciola gigantica to adult F. hepatica excretory/secretory products (Fh-ES) or F. gigantica excretory/secretory products (Fg-ES), respectively. An antibody enzyme-linked immunosorbent assay (Ab-ELISA) was used to determine serum antibody (total Ig, IgG(1), IgM, IgG(2) and IgA) responses. At necropsy, the mean number of flukes recovered was lower in cattle than in sheep. All F. hepatica and F. gigantica infected sheep and cattle showed an increased total Ig levels from 3 to 4 weeks post-infection (wpi). Among isotypes IgG(1) was most dominant while IgM was the earliest (2 wpi) to be detected in both sheep and cattle infected with both F. hepatica and F. gigantica animals. IgG(2) response was early (2 wpi) in sheep infected by F. hepatica but there was no response in sheep infected with F. gigantica. There was a late and strong IgG(2) response in cattle infected with both flukes. The IgA isotype showed an early and a clear biphasic response in sheep with F. hepatica but was less pronounced in F. gigantica infected sheep. While IgA response to Fh-ES was noticed 5 wpi in F. hepatica infected cattle, it appeared much later (21 wpi) in those infected with F. gigantica. The dominance of IgG(1) isotype in infected sheep and cattle suggest an associated Th2 response. This early response to adult Fasciola spp. ES antigen suggests an early exposure to the antigen presumably through the cross-reacting ES products of juvenile flukes. There is clearly difference in IgG(2) isotype response in cattle (resistant) compared to sheep (susceptible). The late IgG(2) response in cattle may suggest late Th1 involvement in bovine cellular responses to adult Fh-ES/Fg-ES.

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.

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.