Circulating Parasite Antigen Research Articles

Consistent Measurement of Parasite-Specific Antigen Levels in Sera of Patients with Neurocysticercosis Using Two Different Monoclonal Antibody (mAb)-Based Enzyme-Linked Immunosorbent Assays.

Monoclonal antibody (mAb)-based enzyme-linked immunosorbent assay (ELISA) is a complementary diagnosis technique for neurocysticercosis (NCC), which detects circulating parasite antigen (Ag) indicative of viable infection and Ag levels that correlate well with the parasite burden. In this study, we compared the performance of two Ag-ELISA techniques for the detection of NCC. We assessed the agreement between our in-house TsW8/TsW5 Ag-ELISA and the widely used B158/B60 Ag-ELISA for measuring T. solium antigen levels in the sera from 113 patients with calcified, parenchymal, and subarachnoid NCC. Concordance was demonstrated evaluating the limits of agreement (LoAs) stratified by the type of NCC. Both ELISA's detected 47/48 (97.8%) subarachnoid NCC cases. In parenchymal and calcified NCC, the B158/B60 Ag-ELISA detected 19/24 (79.2%) and 18/41 (43.9%) cases, while the TsW8/TsW5 Ag-ELISA detected 21/24 (87.5%) and 13/41 (31.7%), respectively. Parenchymal and calcified NCC obtained a perfect agreement (100%), indicating that all sample results were within the predicted LoA, while for subarachnoid NCC, the agreement was 89.6%. The high concordance between the assays was confirmed by Lin's concordance coefficient (LCC = 0.97). Patients with viable parenchymal NCC (LCC = 0.95) obtained the highest concordance between assays, followed by subarachnoid NCC (LCC = 0.93) and calcified NCC (LCC = 0.92). The TsW8/TsW5 Ag-ELISA and B158/B60 Ag-ELISA showed high Ag measurement correlations across diverse types of NCC.

Adult parasite burden and excretion of first-stage larvae of Angiostrongylus vasorum in dogs: Methodologically relevant diagnostic aspects and associations with serological detection of parasite antigen and specific antibodies

Angiostrongylus vasorum is a widely distributed cardiopulmonary parasite of canids in Europe. Clinical signs in dogs can be highly variable and diagnostically challenging. A correct and early diagnosis is hence indispensable to adequately manage affected patients. First-stage larvae (L1) are excreted in the faeces of definitive hosts and conventionally identified using the Baermann technique. Moreover, ELISAs for the detection of circulating antigen and specific antibodies have been presented. The current study aimed at i) quantitatively assessing larval migration in the Baermann funnel after 12 h and 24 h; ii) investigating the influence of sample storage at 4 °C over the course of three days on the number of detected L1; iii) evaluating potential associations of adult worm burdens with larval shedding in dogs and ELISA optical density (OD) values for circulating parasite antigen and specific antibodies. Faecal samples were obtained from naturally infected dogs (n = 21) and Baermann funnels were set up in duplicate over the course of four consecutive days (days 0–3) starting with the day of sample collection. Funnels were harvested on days 1–4 after 12 and 24 h, respectively, and the number of L1 per gram faeces (LPG) was determined. The LPG did not differ between larval harvest after 12 h from harvest after 24 h. Storage of faecal samples at 4 °C for two and three days entailed a considerable decrease in LPG. Adult worm burdens and larval excretion data from previous experiments demonstrated a correlation between worm burden and LPG. In contrast, no correlations between worm burden and the level of parasite antigen and specific antibody OD values, respectively, were identified. Thus, OD values of both antigen and antibody ELISA did not allow for conclusions on infection intensity reflected by the number of adult parasites. For the detection of L1 in faeces, 12 or 24 h of larval migration time was not discriminating for A. vasorum positivity. Thus, early processing of faecal samples is essential, since larval detection and hence sensitivity of the approach considerably decreased over the course of three days of storage. Therefore, the common recommendation to collect faecal samples for three consecutive days and to subsequently analyse them needs to be reconsidered. The results of this study can be readily translated into precise recommendations for daily practice to adequately assess A. vasorum infected dogs.

Porcine model of neurocysticercosis by intracarotid injection of Taenia solium oncospheres: Dose assessment, infection outcomes and serological responses.

BackgroundNeurocysticercosis (NCC) is the infection of the human central nervous system (CNS) by Taenia solium larvae that cause significant neurological morbidity. Studies on NCC pathophysiology, host-parasite interactions or therapeutic agents are limited by the lack of suitable animal models. We have previously reported that carotid injection of activated T. solium oncospheres directs parasites into the CNS and consistently reproduces NCC. This study assessed the minimal dose required to consistently obtain NCC by intracarotid oncosphere injection and compared antigen and antibody response profiles by dose-group.Methods/Principal findingsThree groups of pigs were infected with either 2500 (n = 10), 5000 (n = 11), or 10000 (n = 10) oncospheres. Two pigs died during the study. Necropsy exam at day 150 post-infection (PI) demonstrated viable NCC in 21/29 pigs (72.4%), with higher NCC rates with increasing oncosphere doses (4/9 [44.4%], 9/11 [81.8%] and 8/9 [88.9%] for 2500, 5000, and 10000 oncospheres respectively, P for trend = 0.035). CNS cyst burden was also higher in pigs with increasing doses (P for trend = 0.008). Viable and degenerated muscle cysticerci were also found in all pigs, with degenerated cysticerci more frequent in the 2500 oncosphere dose-group. All pigs were positive for circulating parasite antigens on ELISA (Ag-ELISA) from day 14 PI; circulating antigens markedly increased at day 30 PI and remained high with plateau levels in pigs infected with either 5000 or 10000 oncospheres, but not in pigs infected with 2500 oncospheres. Specific antibodies appeared at day 30 PI and were not different between dose-groups.Conclusion/SignificanceIntracarotid injection of 5000 or more oncospheres produces high NCC rates in pigs with CNS cyst burdens like those usually found in human NCC, making this model appropriate for studies on the pathogenesis of NCC and the effects of antiparasitic treatment.

Immunodiagnostic monoclonal antibody-based sandwich ELISA of fasciolosis by detection of Fasciola gigantica circulating fatty acid binding protein.

Up to now, parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Hence, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. In the present study, a monoclonal antibody (MoAb) against recombinant Fasciola gigantica fatty acid binding protein (rFgFABP) has been produced. As well, a reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) has been developed for the detection of circulating FABP in the sera of mice experimentally and cattle naturally infected with F. gigantica. MoAb 3A3 and biotinylated rabbit anti-recombinant FABP antibody were selected due to their high reactivities and specificities. The lower detection limit of sandwich ELISA was 5 pg mL-1, and no cross-reaction with other parasite antigens was observed. This assay could detect F. gigantica infection from day 1 post infection. In experimental mice, the sensitivity, specificity and accuracy of this assay were 93·3, 100 and 98·2%, while in natural cattle they were 96·7, 100 and 99·1%. Hence, this sandwich ELISA method showed high efficiencies and precisions for diagnosis of fasciolosis by F. gigantica.

Immunodiagnosis of Fasciola gigantica Infection Using Monoclonal Antibody-Based Sandwich ELISA and Immunochromatographic Assay for Detection of Circulating Cathepsin L1 Protease.

BackgroundTropical fasciolosis caused by Fasciola gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Africa and Asia including Thailand. Parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Therefore, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden.MethodsIn this study, we have produced a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1 (rFgCatL1), and developed both sandwich enzyme-linked immunosorbent assay (sandwich ELISA) and immunochromatographic (IC) test for rapid detection of circulating cathepsin L1 protease (CatL1) in the sera from mice experimentally and cattle naturally infected with Fasciola gigantica. MoAb 4E3 and biotinylated rabbit anti-recombinant CatL1 antibody were selected due to their high reactivities and specificities.ResultsThe lower detection limits of sandwich ELISA and IC test were 3 pg/ml and 0.256 ng/ml, respectively. Sandwich ELISA and IC test could detect F. gigantica infection from day 1 to 35 post infection. In experimental mice, the sensitivity, specificity and accuracy were 95%, 100% and 98.6% (for sandwich ELISA), and 93%, 100% and 98.2% (for IC test), while in natural cattle they were 98.3%, 100% and 99.5% (for sandwich ELISA), and 96.7%, 100% and 99.1% (for IC test).ConclusionsThese two assay methods showed high efficiencies and precisions for diagnosis of fasciolosis by F. gigantica.

What is the mechanism of action of praziquantel and how might resistance strike?

Schistosomiasis, also known as bilharzia, is a neglected tropical disease affecting over 249 million people of which approximately 45% are school-aged children. Although there are significant numbers of people with the disease in Asia and South America, over 90% of those infected reside in Africa. The Global Burden of Disease Study 2010 estimated 11,700 deaths and 2,986,000 years lost to disability due to schistosomiasis in that year [1,2]. In 2012, the World Health Organization produced a ‘roadmap’ whose ultimate goal was the “...elimination of NTDs or reductions in their impacts to levels at which they are no longer considered public-health problems” [3]. Schistosomiasis was listed as a disease for potential elimination in the Eastern Mediterranean, Caribbean, Indonesia and Mekong River basin by 2015 and Brazil by 2020. Understandably, a more conservative note was sounded for Sub-Saharan Africa where worry over availability of medication was cited as the reason for failing to schedule elimination by 2020. The best-case scenario that could be made was with widespread availability of medication and strong political support schistosomiasis could be “...eliminated as a ‘public health problem’ in multiple countries in Africa by 2020.” The causative agents of schistosomiasis are digenetic trematode worms belonging the genus Schistosoma with Schistosoma mansoni, S. japonicum and S. haematobium being responsible for the majority of cases. Unfortunately, there is no vaccine available for the treatment of human schistosomiasis nor is the development of one on the horizon. Praziquantel (PZQ) is the only widely available drug for treatment of the disease and is thus at the heart of all control programs. It has been available for over 30 years yet the mechanism by which it kills schistosomes remains unknown and worries persist over the potential for mass administration campaigns to eventually diminish its usefulness. Here, we provide an introduction to schistosomiasis and discuss our current understanding of the mechanism of action of PZQ and the potential for widespread drug resistance.

Aptamer Based, Non-PCR, Non-Serological Detection of Chagas Disease Biomarkers in Trypanosoma cruzi Infected Mice

Chagas disease affects about 5 million people across the world. The etiological agent, the intracellular parasite Trypanosoma cruzi (T. cruzi), can be diagnosed using microscopy, serology or PCR based assays. However, each of these methods has their limitations regarding sensitivity and specificity, and thus to complement these existing diagnostic methods, alternate assays need to be developed. It is well documented that several parasite proteins called T. cruzi Excreted Secreted Antigens (TESA), are released into the blood of an infected host. These circulating parasite antigens could thus be used as highly specific biomarkers of T. cruzi infection. In this study, we have demonstrated that, using a SELEx based approach, parasite specific ligands called aptamers, can be used to detect TESA in the plasma of T. cruzi infected mice. An Enzyme Linked Aptamer (ELA) assay, similar to ELISA, was developed using biotinylated aptamers to demonstrate that these RNA ligands could interact with parasite targets. Aptamer L44 (Apt-L44) showed significant and specific binding to TESA as well as T. cruzi trypomastigote extract and not to host proteins or proteins of Leishmania donovani, a related trypanosomatid parasite. Our result also demonstrated that the target of Apt-L44 is conserved in three different strains of T. cruzi. In mice infected with T. cruzi, Apt-L44 demonstrated a significantly higher level of binding compared to non-infected mice suggesting that it could detect a biomarker of T. cruzi infection. Additionally, Apt-L44 could detect these circulating biomarkers in both the acute phase, from 7 to 28 days post infection, and in the chronic phase, from 55 to 230 days post infection. Our results show that Apt-L44 could thus be used in a qualitative ELA assay to detect biomarkers of Chagas disease.

Parasite antigen in serum predicts the presence of viable brain parasites in patients with apparently calcified cysticercosis only.

Computed tomography (CT) remains the standard neuroimaging screening exam for neurocysticercosis, and residual brain calcifications are the commonest finding. Magnetic resonance imaging (MRI) is more sensitive than CT but is rarely available in endemic regions. Enzyme-linked immunoelectrotransfer blot (EITB) assay uses antibody detection for diagnosis confirmation; by contrast, enzyme-linked immunosorbent assay (ELISA) antigen detection (Ag-ELISA) detects circulating parasite antigen. This study evaluated whether these assays predict undetected viable cysts in patients with only calcified lesions on brain CT. Serum samples from 39 patients with calcified neurocysticercosis and no viable parasites on CT were processed by Ag-ELISA and EITB. MRI was performed for each patient within 2 months of serologic testing. Conservatively high ELISA and EITB cutoffs were used to predict the finding of viable brain cysts on MRI. Using receiver operating characteristic-optimized cutoffs, 7 patients were Ag-ELISA positive, and 8 had strong antibody reactions on EITB. MRI showed viable brain cysts in 7 (18.0%) patients. Patients with positive Ag-ELISA were more likely to have viable cysts than Ag-ELISA negatives (6/7 vs 1/32; odds ratio, 186 [95% confidence interval, 1-34 470.0], P < .001; sensitivity 85.7%, specificity 96.9%, positive likelihood ratio of 27 to detect viable cysts). Similar but weaker associations were also found between a strong antibody reaction on EITB and undetected viable brain cysts. Antigen detection, and in a lesser degree strong antibody reactions, can predict viable neurocysticercosis. Serological diagnostic methods could identify viable lesions missed by CT in patients with apparently only calcified cysticercosis and could be considered for diagnosis workup and further therapy.

USO DEL ELISA DE CAPTURA DE ANTÍGENO EN EL MONITOREO DE PORCINOS INFECTADOS NATURALMENTE CON Cysticercus cellulosae Y TRATADOS CON OXFENDAZOL

Taenia solium cysticercosis is a serious public health problem in developing countries around the world. At the present time, treatment with anthelmintic drugs is possible, but once applied the treatment the monitoring of the animal is limited by the high cost of the radiological techniques. Moreover, the available immunodiagnostic assays are not useful for monitoring because they detect the presence of antibodies against C. cellulosae, which remain in circulation after cysts are dead. The objective of the present study was to demonstrate the usefulness of a monoclonal antibody-based capture ELISA (AgELISA) in the monitoring of the persistence of circulating parasite antigens in animals treated against cysticercosis. For this purpose, serum samples corresponding to the monitoring of pre and post treatment of 6 pigs, naturally infected with cysticercosis and treated with oxfendazol, were evaluated. Necropsy data and evagination records were also used. It was found that the time in which the antigen titles were lower to the cut off OD values occurred on day 59 post treatment and that the mean survival time was 71.5 (59.3-83.8) days. These results seem to demonstrate that the Ag-ELISA would allow the quantitative monitoring of the behavior and persistence of circulating antigens in naturally infected animals that are treated with an antiparasitic drug. However, the monitoring of additional treated pigs are required to obtain conclusive results.

Cross-reactions of sera from dogs infected with Angiostrongylus vasorum in commercially available Dirofilaria immitis test kits

BackgroundDirofilaria immitis and Angiostrongylus vasorum are both important potentially fatal canine nematodes with overlapping endemic areas, especially in Europe. The preadult and adult stages of both species are living in the Arteria pulmonalis and the right heart, and diagnostically detectable circulating parasite antigens have been demonstrated for both species. For the detection of D. immitis infections, a variety of commercial tests have been developed, however, they have not been evaluated for cross-reactions against circulating antigens of A. vasorum.MethodsIn this study, potential cross-reactions of sera from 16 dogs, which were experimentally infected with A. vasorum and which had circulating antigens as confirmed by a species-specific ELISA, were evaluated for the detection of A. vasorum antigen in six commercially available D. immitis test kits.ResultsIn three fast tests (Witness® Dirofilaria, SensPERT® Canine Heartworm, SNAP® 4Dx® Plus), all sera were negative. One fast membrane ELISA (SNAP® HTWM RT Test) was positive with four sera (25%), and one serum delivered a non-valid result twice. In the PetChek® HTWM PF Test, depending on the interpretation protocol, 5 or 8 dogs (31.2 – 50%) were positive. With the DiroCHEK®-ELISA, a single A. vasorum-infected dog (6.2%) tested positive.ConclusionsDue to potential cross-reactions with A. vasorum in commercially available test kits for the detection of D. immitis antigen, the simultaneous use of highly specific diagnostic methods for the differentiation of these two canine heart worms is recommended.

Prevalence of porcine cysticercosis and associated risk factors in Homa Bay District, Kenya

BackgroundTaenia solium is an important zoonosis in many developing countries. Cysticercosis poses a serious public health risk and leads to economic losses to the pig production industry. Due to scarcity of data on the epidemiology of porcine cysticercosis in Kenya, the present study was conducted to determine the prevalence and risk factors for porcine cysticercosis within Homa Bay district. A cross-sectional survey was carried out in 2010, and a total of 392 pigs were recruited in a household survey, with all being tested by ante-mortem lingual palpation (together with questionnaire data on pig production, occurrence and transmission of porcine cysticercosis, risk factors and awareness of porcine cysticercosis collected from the households from which pigs were sampled). Sufficient serum was collected from 232 of the pigs to be tested for the presence of circulating parasite antigen using a monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (Ag-ELISA).ResultsSeventy six pigs were found positive by the Ag-ELISA (32.8%, 95% C.I. 26.8-39.2%), while by tongue inspection cysticerci were detected in 22/ 392 pigs (5.6% 95% C.I. 3.6-8.4%).The most important risk factor for porcine cysticercosis in the Homa Bay area was for pigs to belong to a farm where latrine use by members of the household was not evident (OR = 1.9, 95% CI = 1.13–2.37).ConclusionThe present findings indicate that porcine cysticercosis is endemic in Homa Bay District, and that latrine provision, in conjunction with free-range pig keeping contributes significantly to porcine cysticercosis transmission.

Cysticercus cellulosae antigens in the serodiagnosis of neurocysticercosis

Neurocysticercosis (NCC) is difficult to diagnose clinically because of its varied clinical presentation. However, an accurate diagnosis is possible only after suspicion on epidemiological grounds, proper interpretation of the clinical data, analysis of the findings on imaging studies, and specific immunological tests on the serum and cerebrospinal fluid (CSF). The diagnosis of NCC by any single parameter thus continues to remain difficult. In the past, detection of NCC was based on autopsy studies and histological confirmation. In recent times, the advent of imaging methods such as computed tomography and magnetic resonance imaging have provided excellent non-invasive tools for easy detection of NCC. Nevertheless, an imaging technique of the brain, although useful, is not considered as a gold standard for the diagnosis of NCC. Serological tests are being increasingly used in adjunct with imaging techniques, to aid the diagnosis of NCC. Immunodiagnostic techniques include detection methods for specific antibodies and for circulating parasite antigens in the serum and CSF. Currently, many of the immunodiagnostic tests, including the enzyme-linked immunosorbent assay and enzyme immunotransfer blot, use purified native antigens for the immunodiagnosis of NCC. Nevertheless, the main problem with the use of native cysticercal antigens is that the native proteins often show cross reactions with sera from humans infected with other parasites. The preparation of native antigens also demand a constant supply of parasitic material from the intermediate host pig. In order to overcome the problems in using native antigens, the recombinant antigens or synthetic peptides, which can be produced under stable conditions, are being evaluated for the serodiagnosis of NCC.

Prevalence and Risk Factors of Porcine Cysticercosis in Angónia District, Mozambique

Taenia solium is an important zoonosis in many developing countries. Cysticercosis poses a serious public health risk and incurs sizeable economic losses to pig production. Because data on the epidemiology of porcine cysticercosis in Mozambique are scarce, the present study was conducted to determine the prevalence and risk factors for porcine cysticercosis. A cross-sectional survey was carried out in 11 villages in Angónia district, Tete province in northwestern Mozambique. Between September and November, 2007, a total of 661 pigs were tested serologically and examined by tongue inspection. Serum samples were tested for the presence of circulating parasite antigen using a monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (Ag-ELISA). In addition, a questionnaire survey to collect information on pig production, occurrence and transmission of porcine cysticercosis, risk factors and awareness of porcine cysticercosis was conducted in the selected households from which pigs were sampled. Two hundred thirty-one samples (34.9%) were found positive by the Ag-ELISA, while by tongue inspection on the same animals cysticerci were detected in 84 pigs (12.7%). Increasing age (OR = 1.63; 95% CI = 1.13–2.37) and free-range pig husbandry system (OR = 3.81; 95% CI = 2.08–7.06) were important risk factors for porcine cysticercosis in the district. The present findings indicate that porcine cysticercosis is endemic in the region, and that increasing pig age and pig husbandry practices contribute significantly to porcine cysticercosis transmission. Further epidemiological studies on the prevalence and transmission of porcine cysticercosis in rural communities in Mozambique are needed to enable collection of more baseline data and implementation of effective control strategies within the country.

Age-related infection and transmission patterns of human cysticercosis

Neurocysticercosis is recognised as an important but neglected cause of epilepsy in developing countries where the parasite occurs. Data on the transmission dynamics of the parasite in endemic areas are scarce. Individuals living in these areas are likely to be highly exposed to the parasite, but relatively few of them develop active infections. The present study aimed to describe and gain insights into changes in antibody responses and infection patterns related to age and/or gender in a south Ecuadorian rural population by combining antibody and antigen serological data with demographic characteristics. In 25% of the population, antibodies to Taenia solium cysticerci were detected whilst 2.9% had circulating parasite antigens. The proportion of antibody-positive individuals increased significantly until the age of 40 years to become stable in older individuals. A rule-based simulation model was developed to explain these variations and to reflect the dynamics of exposure to, and transmission of, the parasite. In contrast, the proportion of people presenting circulating parasite antigens, reflecting an active infection, was significantly higher in people older than 60 years. Immunosenescence could explain such an observation since a weaker immune system in the elderly would facilitate the establishment and maintenance of viable cysticerci compared with fully immunocompetent younger individuals. This work points out the role of the immune system in the development of cysticercosis within an exposed population and highlights new essential issues in understanding the transmission dynamics of the parasite, its incidence and the resulting immunological response at a population level.

Production of Monoclonal Antibodies Against the 8 kDa Subunit ofEchinococcus granulosusAntigen B (EgAgB8/2) Using DNA Immunization

Cystic echinococcosis (CE), an endemic cosmopolitan zoonotic helminthic disease caused by the larval stage of Echinococcus granulosus, lacks reliable diagnostic tools that fulfill the criteria of high sensitivity and specificity. Antigen B (AgB), a thermostable lipoprotein that constitutes a considerable fraction of the cystic hydatid fluid (HF), is being considered as a suitable source for vaccination and immunodiagnosis of CE due to its high specificity. Genetic immunization was used to immunize BALB/c mice with the second subunit of antigen B (EgAgB8/2) for the production of monoclonal antibodies (MAb). Fusion products between the spleen cells and myeloma cells produced six MAbs of the following isotypes: IgG2a (two clones), IgG2b (three clones), and IgM (one clone). The MAbs were tested for their specificity to crude sheep hydatid fluid (CSHF) versus other antigens prepared from other helminthic parasites including Toxocara canis, Acanthocheilonema viteae, Fasciola hepatica, Schistosoma mansoni, and Taenia. Five MAbs reacted with E. granulosus antigens, one showed cross reactivity with S. mansonia antigens, and one showed a high reactivity with E. granulosus but was cross reactive with all helminthic antigens tested. Using SDS-PAGE and immunoblotting under reducing conditions, all MAbs identified the four AgB subunits with molecular weights of 8, 16, 24, and 36 kDa. Further work on the specificity and sensitivity of these MAbs as well as their use in detecting circulating parasite antigens and in antigen purification will be assessed in future studies.

Immunoparasitological evaluation of Trichinella spiralis infection during human pregnancy: a small case series

The clinical background, vertical transmission of Trichinella spiralis and helminthotoxic activity of sera were evaluated in four pregnant women infected at different gestation times. Parasitological and immunological parameters were analyzed. Patients presented clinical symptoms and a seroconversion pattern typical of Trichinella infection, independently of the gestation trimester. All patients, including one patient treated with mebendazole, gave birth to healthy infants at full term. Studies performed in placentas and umbilical cords by artificial digestion and/or immunofluorescence did not reveal the presence of parasites or parasite antigens. Specific antibodies were found in placental extraction and maternal sera by immunofluorescence, ELISA and/or immunoelectrotransfer blot assay. Specific IgG, IgE and IgA were found in the umbilical cord sera. One umbilical cord demonstrated immune-complexed specific IgM and circulating parasite antigens were found in the corresponding infant up to 10 months after birth. Patients' sera were able to kill newborn larvae in cytotoxicity assays, even in the absence of specific antibodies; this effect was abrogated by mifepristone. The results suggest that in human trichinellosis during pregnancy there is an enhanced helminthotoxicity to newborn larvae, dependent in part on progesterone, leading to a mild or moderate course of the infection. The results also indicate that the transplacental passage of migrating larvae is possible.

Estudo histológico e imunoistoquímico do sistema nervoso central de cães naturalmente infectados por Leishmania (Leishmania) chagasi

O objetivo do presente estudo foi caracterizar as alterações histopatológicas e detectar por imunoistoquímica, a presença de formas amastigotas de Leishmania no sistema nervoso central de cães com e sem sintomatologia neurológica da doença. Foram utilizados dois grupos de animais; o primeiro foi composto de 18 cães com leishmaniose visceral sem evidência clínica de envolvimento neurológico, e o segundo, composto de 21 cães com leishmaniose visceral e sintomas neurológicos. As mais freqüentes alterações histopatológicas encontradas no sistema nervoso central de cães de ambos os grupos foram degeneração neuronal com neuronofagia, gliose, leptomeningite, congestão vascular, presença de infiltrado linfoplasmocitário perivascular e áreas de microhemorragia. Não foram observadas marcações antigênicas para formas amastigotas de Leishmania em nenhum fragmento do sistema nervoso central de cães de ambos os grupos; entretanto, a maioria deles apresentava marcação em parede de vasos sangüíneos, o que sugere a presença de antígenos parasitários circulantes.

&lt;i&gt;Taenia saginata&lt;/i&gt; cysticercosis in Oyo State, Nigeria

A retrospective analysis of abattoir records in Oyo State of Nigeria showed that 0.45% the total 661,048 cattle carcasses inspected between 1989 and 1998 had Cysticercus bovis. Participatory meat inspection at Moniya slaughter slab in Ibadan between August and October, 2000 showed that 1.76% of the 1,413 carcasses examined were infected. The predilection sites were tongue, heart, masseter muscle, liver and lung in that order with multiple organs affected in 28% of the carcasses. Seventy-two percent of the infected carcasses had a total of 160 viable cysts. A serological survey was conducted to detect circulating parasite antigens in serum samples of trade cattle using a monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (Ag-ELISA). Thirty-four (12.8%) of the 265 serum samples screened were positive while cysticerci cysts were detected in only 9 (3.4%) of the corresponding carcasses during routine meat inspection. Sero-prevalence was positively correlated with ages of the animals while sex and breed had no significant effect, respectively. Improved meat inspections at abattoirs and with the adoption of the highly sensitive ante-mortem serological diagnosis technique will obviously result in the establishment of the true prevalent status of the infection in Nigeria and also facilitate effective control and eradication of the disease in man and animals. Keywords: Taenia saginata, cattle, Ag-ELISA Tropical Veterinarian Vol. 22(3&4) 2004: 113-117

Evidence for high seroprevalence of Taenia solium cysticercosis in individuals from three rural communities in Venezuela

A serological study was undertaken in 1998 to evaluate levels of Taenia solium cysticercosis in 3 rural Venezuelan communities. Infection with viable metacestodes was diagnosed with a trapping enzyme-linked immunosorbent assay (ELISA) that detects a secreted product of viable parasites. Anti-metacestode antibodies were assayed by ELISA using T. solium vesicular fluid as antigen. A total of 1254 sera was collected from 3 communities (Canoabo, Sanare, and Rio Tocuyo) where previous studies had suggested the presence of T. solium. Our results demonstrate an unusually high seroprevalence of cysticercosis, indicating an attendant risk of transmitting the disease to other areas. The seroprevalence of infection with viable cysts, as indicated by detection of circulating parasite antigen, was 9.1% in Canoabo, 6.1% in Sanare, and 5.7% in Rio Tocuyo. The corresponding frequency of antibodies to T. solium cyst antigens was 36.5% in Canoabo, 36.5% in Sanare, and 4% in Rio Tocuyo. As these communities are probably representative of many others in Venezuela, T. solium cysticercosis may be a significant public health problem and more work is certainly indicated. An important finding was that local knowledge of the disease and its transmission do not necessarily guarantee diminished disease prevalence, indicating a lack of appropriate vigilance towards disease control.

The prevalence of porcine cysticercosis in Eastern and Southern provinces of Zambia.

The objective of this study was to determine the prevalence and importance of porcine cysticercosis in rural areas of Zambia. The study involved an abattoir survey of 1316 pigs at a slaughter slab in Lusaka and two field surveys in villages in Southern and Eastern provinces. Lingual examination of live pigs and visual inspection of their carcass as well as blood sampling for measuring circulating parasite antigen by enzyme-linked immunosorbent assay (Ag-ELISA) were used as parameters to measure infection. In the field surveys, a questionnaire was administered to every household whose pigs were examined to obtain information on pig husbandry practices and to study risk factors for the infection. Out of the 1316 pigs examined at the slaughter slab, 143 (10.9%) and 271 (20.6%) were positive by lingual examination and meat inspection, respectively. Most of the pigs were very heavily infected with predominantly live cysts. The field surveys revealed that eight (8.2%) out of 98 pigs from Southern province and eight (5.2%) out of 151 pigs from Eastern province were positive for cysticercosis by tongue palpation. Using the Ag-ELISA 20 (20.8%) and 14 (9.3%) pigs were positive in Southern and Eastern provinces, respectively. The questionnaire survey revealed poor pig husbandry practices, absence of meat inspection and control, poor knowledge of the disease and poor sanitation in the surveyed villages. The prevalence of pig cysticercosis found in this study ranks among the highest in the southern African region, in Africa and in the world. The current study suggests the presence of human tapeworm carriers and a high risk of human cysticercosis in the surveyed areas as well as in urban centres where pigs from rural areas are increasingly sold, slaughtered and consumed.