A Novel Disease of Big-Leaf Mahogany Caused by Two Fusarium Species in Mexico.

Big-leaf mahogany (Swietenia macrophylla) is valued for its high-quality wood and use in urban landscapes in Mexico. During surveys of mango-producing areas in the central western region of Mexico, symptoms of malformation, the most important disease of mango in the area, were observed on big-leaf mahogany trees. The objectives of this research were to describe this new disease and determine its cause. Symptoms on big-leaf mahogany at four sites in Michoacán, Mexico resembled those of the vegetative phase of mango malformation, including compact, bunched growth of apical and lateral buds, with greatly shortened internodes and small leaves that curved back toward the supporting stem. Of 163 isolates that were recovered from symptomatic tissues, most were identified as Fusarium pseudocircinatum (n = 121) and F. mexicanum (n = 39) using molecular systematic data; two isolates represented unnamed phylospecies within the F. incarnatum-equiseti species complex (FIESC 20-d and FIESC 37-a) and another was in the F. solani species complex (FSSC 25-m). However, only F. mexicanum and F. pseudocircinatum induced malformation symptoms on 14-day-old seedlings of big-leaf mahogany. The results indicate that F. mexicanum and F. pseudocircinatum, previously reported in Mexico as causal agents of mango malformation disease, also affect big-leaf mahogany. This is the first report of this new disease and the first time that F. mexicanum was shown to affect a host other than mango.

Two glutathione transferase isoforms isolated from juvenile cysts of Taenia crassiceps: identification, purification and characterization.

We identified and characterized the first two glutathione transferases (GSTs) isolated from juvenile cysts of Taenia crassiceps (EC 2.5.1.18). The two glutathione transferases (TcGST1 and TcGST2) were purified in a single-step protocol using glutathione (GSH)-sepharose chromatography in combination with a GSH gradient. The specific activities of TcGST1 and TcGST2 were 26 U mg-1 and 19 U mg-1, respectively, both at 25°C and pH 6.5 with 1-chloro-2,4-dinitrobenzene (CDNB) and GSH as substrates. The Km(CDNB) and Kcat(CDNB) values for TcGST1 and TcGST2 (0.86 µm and 62 s-1; 1.03 µm and 1.97 s-1, respectively) and Km(GSH) and Kcat(GSH) values for TcGST1 and TcGST2 (0.55 µm and 11.61 s-1; 0.3 µm and 32.3 s-1, respectively) were similar to those reported for mammalian and helminth GSTs. Mass spectrometry analysis showed that eight peptides from each of the two parasite transferases were a match for gi|29825896 glutathione transferase (Taenia solium), confirming that both enzymes are GSTs. The relative molecular masses were 54,000 ± 0.9 for the native enzymes and 27,500 ± 0.5 for the enzyme subunits. Thus, TcGST1 and TcGST2 are dimeric proteins. Optimal TcGST1 and TcGST2 activities were observed at pH 8.5 in the range of 20-55°C and pH 7.5 at 35-40°C, respectively. TcGST1 and TcGST2 were inhibited by cibacron blue (CB), bromosulphophthalein (BST), rose bengal (RB), indomethacin and haematin (Hm) with 50% inhibitory concentrations (IC50) in the µm range. TcGST1 was inhibited in a non-competitive manner by all tested inhibitors with the exception of indomethacin, which was uncompetitive. The discovery of these new GSTs facilitates the potential use of T. crassiceps as a model to investigate multifunctional GSTs.

Evaluation of the non-catalytic binding function of Ts26GST a glutathione transferase isoform of Taenia solium.

Taenia solium glutathione transferase isoform of 26.5 kDa (Ts26GST) was observed to bind non-catalytically to porphyrins, trans-trans-dienals, bile acids and fatty acids, as assessed by inhibition kinetics, fluorescence spectroscopy and competitive fluorescence assays with 8-anilino-1-naphthalene sulfonate (ANS). The quenching of Ts26GST intrinsic fluorescence allowed for the determination of the dissociation constants (KD) for all ligands. Obtained data indicate that Ts26GST binds to all ligands but with different affinity. Porphyrins and lipid peroxide products inhibited Ts26GST catalytic activity up to 100% in contrast with only 20-30% inhibition observed for bile acids and two saturated fatty acids. Non-competitive type inhibition was observed for all enzyme inhibitor ligands except for trans-trans-2,4-decadienal, which exhibited uncompetitive type inhibition. The dissociation constant value KD = 0.7 µM for the hematin ligand, determined by competitive fluorescence assays with ANS, was in good agreement with its inhibition kinetic value Ki = 0.3 µM and its intrinsic fluorescence quenching KD = 0.7 µM. The remaining ligands did not displace ANS from the enzyme suggesting the existence of different binding sites. In addition to the catalytic activity of Ts26GST the results obtained suggest that the enzyme exhibits a ligandin function with broad specificity towards nonsubstrate ligands.

Limits of the therapeutic properties of synthetic S3Pvac anti-cysticercosis vaccine.

The S3Pvac synthetic vaccine, composed of three peptides (GK1, KETc1 and KETc12) effectively protects against cysticercosis under experimental and field conditions. Additionally, S3Pvac vaccine can effectively damage early-established cysticerci in experimentally lightly infected young pigs. This study was designed to explore if also fully-developed cysticerci that eluded immunity induced by the infection can be damaged by S3Pvac-induced immunity in naturally, heavily infected adult pigs. Fourteen pigs identified as cysticercotic by tongue inspection from rural communities were purchased and moved to controlled conditions in the Faculty of Veterinary Medicine. Half of these pigs were treated once a month three times with S3Pvac plus saponin, and the other half received only saponin (controls). Twelve months later pigs were euthanized, and the number of cysticerci, their macro and microscopic status and their capacity to transform into tapeworms were determined. S3Pvac failed to damage fully-developed muscle cysticerci of naturally, heavily infected adult pigs. To explore possible factors involved in the failure of the therapeutic capacity pooled sera from control and treated cysticercotic pigs were added to mice mononuclear peripheral cells. Pooled sera from non-infected pigs were also tested. Sera from control and treated infected pigs almost completely suppressed the T cell proliferative responses, pointing to the presence of suppressor factors. In conclusion, S3Pvac vaccine failed to damage fully-developed cysticerci in pigs in which a host parasite relationship had evolved after months of infection with immunological implications.

First Report of Powdery Mildew on Carrot Caused by Erysiphe heraclei in Michoacan, Mexico.

Carrot (Daucus carota L. subsp. sativus (Hoffm.) Arcang.) is planted as a home-grown vegetable in the central region of Michoacan, Mexico. Powdery mildew was observed on carrot plants cv. Nantesa at several locations near Morelia, Michoacan during March 2009. Affected plants had abundant, white, superficial conidia and mycelium on leaves and stems. All plants at each of five locations surveyed had powdery mildew symptoms with percent foliage coverage ranging from 50 to 80%. Mycelial growth was amphigenous, mainly on the upper leaf surface, covering the whole leaf and with irregular patches on inflorescences and stems. Hyphae were ectophytic with lobed appressoria. Conidiophores presented foot cells 22.5 to 35 (30) × 5.75 to 7 (6.3) µm followed by two cells, one shorter and one longer than the foot cell. Conidia were produced singly, most subcylindric to cylindric, lacked fibrosin bodies, and measured 31.2 to 42 (36.2) × 8.7 to 11.2 (10.5) µm. The teleomorph was not observed. Genomic DNA was extracted from infected leaves; sequences of the internal transcribed spacers (ITS) inclusive of 5.8S rDNA were amplified using previously described primers specific for Erysiphales (3). The ITS sequences shared 100% homology to Erysiphe heraclei specimen VPRI41227 from carrot in Australia (GenBank Accession No. EU371725). On the basis of the morphological characteristics observed and the ITS rDNA sequences, the pathogen was identified as E. heraclei DC. The ITS sequence was deposited in NCBI as Accession No. GU252368. Pathogenicity tests were conducted twice on a total of 10 healthy 8-week-old carrot plants cv. Nantesa. Infected plants were placed in close proximity to healthy plants and maintained in a greenhouse at 27 ± 5°C. Initial signs and symptoms were observed 3 weeks after inoculation and appeared as small, white colonies, which later coalesced and covered most of the foliage. Microscopic examination of the conidia and mycelial morphology matched the originally described pathogen, E. heraclei. Powdery mildew caused by this pathogen has been extensively reported on diverse species and genera of the Apiaceae in Europe and remains one of the most important diseases of carrot (2). The appearance of E. heraclei in diverse regions on a variety of umbelliferous crops indicates that formae speciales have spread, infecting different and specific hosts (1-3). Recently, E. heraclei has been reported on parsley in Puebla, Mexico (4). To our knowledge, this is the first report of E. heraclei causing powdery mildew on carrot in Michoacan, Mexico. This pathogen should be considered as a threat to commercial carrot crops in Mexico. Other crops in the Apiaceae may not be at risk in this area if this powdery mildew is specific for carrots. References: (1) B. J. Aegerter. Page 22 in: Compendium of Umbelliferous Crop Diseases. The American Phytopathological Society, St. Paul, MN, 2002. (2) U. Braun. The Powdery Mildew (Erysiphales) of Europe. Gustav Fischer-Verlag. Jena, Germany, 1995. (3) J. H. Cunnington et al. Australas. Plant Pathol. 32:421, 2003. (4) M. J. Yáñez-Morales et al. Schlechtendalia 19:47, 2009.

Purification, characterization and kinetic properties of the Taenia solium glutathione S-transferase isoform 26.5 kDa.

Glutathione S-transferases are major phase II detoxification enzymes. Taenia solium, a parasite of humans and pigs, is exposed to toxic products. The aim of this work was to purify and characterize a T. solium glutathione S-transferase isoform of 26.5 kDa (SGST26.5) in order to obtain its kinetic parameters. Homogeneous SGST26.5 was obtained by a simple purification procedure. SGST26.5 showed a p I of 7.07, and a native Mr of 60 kDa with 26.5 kDa subunits. The optimum activity for SGST26.5 was found at pH 6.5-7.0 in the range 10-42 degrees C. SGST26.5 had a specific enzyme activity of 78, 7.1, 6.6, and 0.7 microM min(-1) mg(-1) with CDNB, 1,2-dichloro-4-nitrobenzene, 2,4-hexadienal and trans-2-nonenal as substrates, respectively. It also had a kcat/ K(mCDNB)=2.15 x 10(3) M(-1 )s(-1), kcat/ KmGSH)=4.5 x 10(3) M(-1 )s(-1) and Vmax for GSH and CDNB=74 and 77 microM min(-1) mg(-1), respectively. SGST26.5 was inhibited in a noncompetitive form by cibacron blue, bromosulfophthalein and triphenyltin chloride. Inhibition studies as a function of inhibitor concentration show that the enzyme is a homodimer. Bireactant system analysis show that it follows an ordered sequential mechanism.

Evaluation of tongue inspection and serology for diagnosis of Taenia solium cysticercosis in swine: usefulness of ELISA using purified glycoproteins and recombinant antigen.

Evaluation of serology using glycoproteins (GPs) purified by preparative isoelectric focusing (pH 8.8) and recombinant chimeric antigen (RecTs) of Taenia solium was carried out using (1) blood samples on filter papers from pigs infected with different doses of eggs of T. solium in Mexico, (2) serum samples from pigs found infected naturally in Vietnam and Ecuador and (3) serum samples from pigs suspected to be infected with T. solium by tongue inspection in Tanzania. Antibody responses (IgG) were detectable in experimentally infected pigs confirmed harbouring 16 or more cysts at necropsy from 30 days after egg inoculation. One of three pigs naturally infected and harbouring 2.5 cysts/kg muscle and most of pigs harbouring=5.0 cysts/kg were also seropositive by ELISA. Although pigs may be infected with other taeniid species such as Taenia hydatigena, pigs harbouring this parasite were negative in ELISA. Approximately, 76 and 78% of sera from pigs having nodule(s) in the tongue (positive tongue inspection) were serologically positive by both ELISA and immunoblot, respectively. Furthermore, approximately 34 and 18% of sera from pigs having no nodules in the tongue (negative tongue inspection) were also seropositive by ELISA and immunoblot, respectively. ELISA using the two antigens was more sensitive than immunoblot and reliable for differentiation of pigs infected with cysticerci of T. solium from those either uninfected or infected with other taeniid species. Pigs without nodule by tongue inspection should be checked serologically in endemic areas.

Purification of Taenia solium cysticerci superoxide dismutase and myoglobin copurification.

Superoxide dismutase from Taenia solium cysticerci ( Ts SOD) was purified by sequential ion exchange chromatography on quaternary-amino-ethyl-cellulose (QAE) followed by hydrophobic interaction on phenyl sepharose (PS) and chromatofocusing on a polybuffer exchanger 94 (PBE). Ts SOD is a 30 kDa molecular weight dimeric enzyme with 15 kDa monomers. It is partially negative, hydrophilic, with 6.3 isoelectric point and has 2,900 U/mg activity. Bovine erythrocyte SOD antibodies cross react with Ts SOD. This enzyme is 80% inhibited by 10 mM of KCN suggesting that it has a Cu/Zn active site. Furthermore, Ts SOD totally loses its activity at 100 degrees C for 4 min. The first 25 amino acids from the Ts SOD N-terminal are (M K A V X V M R G E E G V K G V V H F T Q A G D A). This sequence is 76% similar to the Schistosoma mansoni Cu/Zn SOD. By chance, myoglobin (Mb) was also found during the purification process. A 16 kDa band was recognized in immunoblotting by horse heart Mb antibodies in QAE, PS and PBE, the last-mentioned being found at pH 7.0. The first 15 amino acids from the amino terminal group (G L S D G E W Q L V L N V W G) in this 16 kDa protein are identical to several other Mbs which have been reported.

Prevalence and risk of cysticercosis and taeniasis in an urban population of soldiers and their relatives.

To determine markers of Taenia solium transmission and risk factors in an urban community, we studied 1,000 soldiers from a military camp in Mexico City and their relatives. Serum samples were used to detect antigens and antibodies and fecal specimens were examined for Taenia coproantigens and helminth eggs. Prevalences of 12.2% and 5.8% for cysticercosis were found among soldiers and their relatives, respectively. Taeniasis was found in 0.5% and none of the groups, respectively. Relatives of soldiers positive for cysticercosis and taeniasis markers ate more pork from street stores than restaurants or markets compared with relatives of soldiers without these indicators of infection. Also, 12.0% of the relatives of positive soldiers had a history of expelling tapeworm proglottids in the feces in contrast to 3.7% of the family members of the control group. Prevalence values and risk factors in this urban population are similar to those of previous studies performed in rural populations.

Characterization of GP39-42 and GP24 antigens from Taenia solium cysticerci and of their antigenic GP10 subunit.

Diagnosis of neurocysticercosis is performed by Western blotting with an enriched fraction of glycoproteins (LLGP). GP39-42 and GP24 are immunodominant antigens. These antigens were electroeluted and characterized by biochemical methods. When GP39-42 or GP24 were reduced, a band of 10 kDa (named GP10) appeared; this band was also analyzed. The most abundant amino acids in the three GPs were lysine, phenylalanine, asparagine, glycine, and leucine. The amino terminal portion was sequenced, and the following order was obtained for the three GPs: EKNKPKNVAXSTKKGYEYVXEF. The glycan portion was 8.4%, 18.2%, and 18.3% in GP39-42, GP24, and GP10, respectively. The three GPs contained mannose, N-acetyl-D-glucosamine, and galactose. GP39-42 and GP24 seem to be oligomeric forms of GP10. When reduced LLGP was reacted with samples obtained from patients with neurocysticercosis or pigs with cysticercosis, a band corresponding to GP10 was always observed. Furthermore, hyperimmune serum from rabbits immunized with GP39-42 or with GP24 recognized GP10 as well as GP39-42 and GP24. The data obtained in this paper suggest that GP10 might be a useful tool for diagnosis.

Vaccination against Taenia solium cysticercosis in pigs using native and recombinant oncosphere antigens.

Pigs were immunised with antigens derived from Taenia solium oncospheres or with a pool of three recombinant antigens from Taenia ovis, and subsequently challenged with T. solium eggs. The native oncosphere antigens induced 83% protection against viable, and 89% protection against the total number of cysticerci established following the challenge infection. Immunisation with the recombinant T. ovis antigens induced 93% protection against the establishment of viable cysticerci, and 74% protection against the total number of cysticerci. These results, and those achieved elsewhere with Taenia saginata and T. ovis, support the possibility of developing a practical vaccine to assist in the control of transmission of T. solium through pigs.

Taenia solium cysticercosis: immunity in pigs induced by primary infection.

The present study demonstrates that pigs experimentally infected with Taenia solium eggs develop resistance to reinfection that lasts at least five months. Thirteen 2-month-old piglets were infected with eggs of Taenia solium. After 5 months, two pigs were euthanized and five were challenged with eggs from a second tapeworm. Nine months after the first infection, six pigs were challenged with a third tapeworm. All 11 challenged pigs were euthanized 2 months after reinfection. In order to confirm the infectivity of the eggs, several piglets were inoculated with each taenia. Two of the five pigs reinfected after 5 months did not develop metacestodes, two showed few caseous non-infective forms and in the fifth pig, 14% of the metacestodes were vesicular and 86% colloidal and caseous. In the six animals challenged 9 months after the first infection, three were heavily infected with vesicular metacestodes and the other three showed only colloid and caseous forms in muscles. All parasites found in brains were vesicular. We conclude that immunity due to primary infection lasts at least 5 months. At 2 months of infection antigens of 24 and 39-42 kDa were the most frequently recognised. In those pigs with only a few caseous cysts in muscles and/or vesicular ones in brains no antibodies were detected.

ELISA and immunoblot using purified glycoproteins for serodiagnosis of cysticercosis in pigs naturally infected with Taenia solium.

The establishment of reliable serological methods for cysticercosis in pigs is important for the surveillance, control and prevention of taeniosis/cysticercosis in humans as well as in pigs to prevent economic loss. Both ELISA and immunoblot using glycoproteins (GPs) purified by a single step of preparative iso-electric focusing, which are highly useful for human cysticercosis, have been applied for a serological study in pigs naturally infected with Taenia solium. All sera from pigs showed similar responses to those in human cysticercosis. Therefore, it is expected that both ELISA and immunoblots using GPs would be useful in differentiating infected pigs from uninfected ones.

Novel antigens for neurocysticercosis: simple method for preparation and evaluation for serodiagnosis.

Neurocysticercosis (NCC), which is caused by infection with the larval stage of the pork tapeworm (Taenia solium), is now recognized as a major cause of neurologic diseases in countries where the infection is endemic. Migration of persons from these countries is resulting in diagnosis and local transmission in nonendemic countries at increasing rates. In the present study, immunoblotting and an ELISA were carried out using antigens of T. solium cysticerci fractionated by isoelectric focusing and serum samples from patients with NCC, alveolar (AE) or cystic echinococcosis (CE), and other diseases. Immunoblot analysis revealed antigens fractionated by isoelectric focusing (pH 9.2-9.6) either from cyst fluid of T. solium cysticerci or from intact cysts had unique components (glycoproteins) highly specific and sensitive for detection of NCC exclusively. All confirmed NCC serum samples (53 of 53) recognized at least three major bands of 10-26-kD of fractions with pH 9.2-9.6 from either intact cysts or cyst fluid. These bands were not recognized by sera from patients with other parasitic diseases including AE (0 of 34), CE (0 of 36), or other heterologous parasitoses (0 of 77), patients with hepatoma (0 of 19) or sarcoidosis (0 of 11), or sera from healthy controls (0 of 29). The ELISA using the antigens showed the same sensitivity and specificity for differentiation of NCC (53 of 53) from other diseases (0 of 107) or healthy individuals (0 of 29). Both immunoblotting and the ELISA using the fractionated antigens readily differentiated all NCC from AE or CE in a blind test of 29 serum samples of persons with NCC, CE, and AE. Antigens fractionated from cyst fluid of T. solium cysticerci by a simple, single-step isoelectric focusing (pH 9.2-9.6) are highly specific and sensitive for differential serodiagnosis of NCC in immunoblotting and/or an ELISA.

Development and evaluation of a health education intervention against Taenia solium in a rural community in Mexico.

A comprehensive study was undertaken in a rural community in the state of Morelos, Mexico to evaluate health education as an intervention measure against Taenia solium. An educational program was developed to promote recognition and knowledge of the transmission of the parasite and to improve hygienic behavior and sanitary conditions that foster transmission. The effects of educational intervention were evaluated by measuring changes in knowledge and practices and prevalence of human taeniasis and swine cysticercosis before and after the campaign. The health education strategy was implemented with the active participation of the population based on the information obtained from a sociologic study. A questionnaire was designed and used before, immediately after the intervention, and six months later. Statistically significant improvements occurred in knowledge of the parasite, its life cycle, and how it is acquired by humans; however, changes in behavior related to transmission were less dramatic and persistent. The prevalences of cysticercosis in pigs at the start of the education intervention were 2.6% and 5.2% by lingual examination and antibody detection (immunoblot assay), respectively, and approximately one year after the intervention they were 0% and 1.2% (P < 0.05). These changes were accompanied by significant reductions in the reported access of pigs to sources of infection and freedom to roam. We conclude that health education, developed along with community involvement, reduced opportunities for transmission of T. solium in the human-pig cycle.

PIP: Neurocysticercosis is an important health problem in Mexico, as well as in many other countries of Latin America, Asia, and Africa where conditions permit completion of the cestode's life cycle in pigs and humans. A study was conducted in a rural community in the state of Morelos, Mexico, to determine whether health education could be an effective measure against Taenia solium. An educational program was developed with community input to promote recognition and knowledge of the transmission of the parasite and to improve hygienic behavior and sanitary conditions which foster transmission. The effects of the educational intervention were then assessed by measuring changes in knowledge, practices, and the prevalence of human taeniasis and swine cysticercosis before and after the campaign. Statistically significant improvements occurred with regard to knowledge of the parasite, its life cycle, and how it is acquired by humans. However, changes in behavior related to transmission were less marked and persistent. Lingual examination and antibody detection found cysticercosis among 2.6% and 5.2% of pigs, respectively, at the start of the intervention. At approximately 1 year after the intervention, prevalences had declined to 0% and 1.2%. The decline was accompanied by significant reductions in the reported access of pigs to sources of infection and freedom to roam.

Experimental Taenia solium cysticercosis in pigs: characteristics of the infection and antibody response.

Pigs were infected with taeniid eggs to study the susceptibility to infection and reinfection of the animals of mixed breeds and of different ages, the viability and death of the metacestodes in the host tissue, and the antibody response which accompanies these events. Sixteen pigs were infected with Taenia solium eggs for this purpose. At necropsy metacestodes were counted in 2 kg of shoulder muscles and classified as vesicular or caseous, and all the metacestodes in brains were counted and classified. The results show that pigs inoculated at 49 and 60 days of age became infected to different degrees and reacted differently to the presence of parasites. In the brain the metacestodes remain viable for longer periods than in muscles. Enzyme-linked immunosorbent assay (ELISA) showed a significant rise in antibodies after infection, which started to decrease 92 days post-infection (p.i.). Pigs with viable cysts remained seropositive up to the end of the experiment (281 days p.i.). Antibody levels rose further after reinfection or after treatment. The results of Western blot were comparable to those of ELISA. Antigens of 13, 14 and 18 kDa were most frequently recognized in early infections and then started to decrease 92 days p.i., while the antigens of 42, 50 and 24 kDa were recognized during later stages of infection (200 days p.i.). The results suggest that older animals are more resistant to the infection [corrected].

Detection by immunoblot assay of antibodies to Taenia solium cysticerci in sera from residents of rural communities and from epileptic patients in Bali, Indonesia.

Several studies from Bali have indicated the presence of Taenia solium. Relatively little has been reported, however, implicating human exposure to this parasite on Bali based upon the prevalence of anti-T. solium antibodies in asymptomatic and epileptic individuals. This study was conducted to determine by immunoblot assay and ELISA the frequency of anti-cysticercus antibodies in two groups of Balinese. Among 746 residents of four ecologic zones, 94 (13%) were positive by immunoblot. Of 74 epileptic patients from throughout the island, 10 (14%) were positive by immunoblot and 8 (11%) by ELISA; however, only 4 (22%) of the 18 sera positive in either test were positive in both assays. The previously defined high specificity and sensitivity of immunoblot indicates that T. solium cysticercosis is well established in Bali and that a significant amount of epilepsy may be due to neurocysticercosis.

Reactivity in ELISA and dot blot of purified GP24, an immunodominant antigen of Taenia solium, for the diagnosis of human neurocysticercosis.

In this paper we report the purification of GP24, one of the seven specific and highly antigenic Taenia solium glycoproteins previously identified by Western blot (WB) with serum, cerebrospinal fluid (CSF) and saliva samples from patients with neurocysticercosis (NC). GP24 was purified and evaluated in ELISA and dot blot for diagnosis. A lentil-lectin-bound glycoprotein fraction (LL-GP) from T. solium cysticerci was submitted to polyacrylamide gel electrophoresis, and the band that corresponded to GP24 was sliced, minced and electroeluted; an aliquot was used to immunize a rabbit, and the antiserum obtained was analysed by WB against LL-GP fraction; only GP24 was detected. ELISA and dot blot were performed with purified GP24 and serum and CSF samples from patients with NC that were previously positive for GP24 in WB and control samples; the latter were negative, while all NC samples were positive. To test for specificity, purified GP24 was incubated in dot blot against 44 sera from patients with other parasitic diseases; no positive reactions were found. Results indicate that GP24 was adequately purified and retained its reactivity; thus in combination with ELISA or dot blot may facilitate immunodiagnosis of NC.

[Diagnosis of human cysticercosis with ELISA]. / El diagnóstico de la cisticercosis humana por ELISA.

An analysis of immunodiagnostic data for human cysticercosis by ELISA in 1,052 cerebrospinal fluid (CSF) and serum samples obtained directly from patients or their physicians from different hospitals during 1989-1991, as a result of the diagnostic support service that has been offered to the medical community since 1976, was performed. Overall positivity was 18 per cent. This value was slightly lower when only CSF was analysed (14%) and lowest in the case of serum samples (4%). Analysis of the CSF positivity in ELISA with respect to imaging findings yielded a predictive value of 93 per cent while in cases of excision of cysticerci by neurosurgery 67 per cent sensitivity was obtained. The high positive predictive value, the specificity and the sensitivity of ELISA indicate that this assay is useful to confirm the imaging diagnosis of neurocysticercosis when CSF is used or it can be used as a diagnostic alternative when imaging studies are not available.

Community-based epidemiological investigations of cysticercosis due to Taenia solium: comparison of serological screening tests and clinical findings in two populations in Mexico.

We compared a plate enzyme-linked immunosorbent assay (ELISA) with an immunoblot--or enzyme-linked immunoelectrotransfer blot--assay (EITB) for the identification of cases of human neurocysticercosis due to Taenia solium and of risk factors for this disease in two Mexican villages. Findings related to age- and sex-specific seroprevalence, risk factors for transmission, and associated morbidity differed significantly according to the assay used. Rates of EITB positivity were significantly higher among persons with a history of convulsions than among those without such a history (29% vs 8%; P < .05); in contrast, ELISA results were negative for all persons with a history of convulsions. The association of seizures with neurocysticercosis in this population was strengthened by the higher rate of abnormal findings compatible with neurocysticercosis on computed tomography of the brain among individuals with a history of convulsions than among those without such a history (70% vs 14%; P < .001). In summary, EITB was more sensitive and specific than ELISA for epidemiological studies of neurocysticercosis.