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The use of L protein coupled magnetic particles for the concentration and purification of immunoglobulin M (mIgM) monoclonal antibodies against Taenia solium was evaluated. Three concentration methods and different elution times were evaluated and the ratio of particles to the ratio of mIgM was optimized. It is demonstrated that: 1) with the use of magnetic particles, a previous concentration of mIgM is not required, which reduces the manipulation of the antibodies and improves the recovery, 2) the use of a binding buffer can be omitted, since the pH of most cell culture supernatants are neutral, and 3) longer elution times (~ 45 minutes) are needed to increase recovery to a level greater than 80%. The study demonstrates that the use of L protein-coupled magnetic particles is a simple and efficient tool for mIgM concentration and purification.

Babesia duncani is the causative agent of babesiosis in the western United States. The indirect fluorescent antibody (IFA) assay is the diagnostic test of choice for detection of B. duncani-specific antibodies. However, this test requires parasitized red blood cells harvested from infected hamsters, and test results are often difficult to interpret. To simplify serological testing for B. duncani, a proteomics approach was employed to identify candidate immunodiagnostic antigens. Several proteins were identified by electrospray ionization mass spectrometric analysis, and four recombinant protein constructs were expressed and used in a multiplex bead assay (MBA) to detect B. duncani-specific antibodies. Two antigens, AAY83295.1 and AAY83296.1, performed well with high sensitivities and specificities. AAY83295.1 had a higher sensitivity (100%) but lower specificity (89%) than AAY83296.1, which had a sensitivity of 90% and a specificity of 96%. Combining these two antigens did not improve the performance of the assay. This MBA could be useful for diagnosis, serosurveillance, and blood donor screening for B. duncani infection.

Babesia duncani is the causative agent of babesiosis in the western United States. The indirect fluorescent antibody (IFA) assay is the diagnostic test of choice for detection of B. duncani-specific antibodies. However, this test requires parasitized red blood cells harvested from infected hamsters, and test results are often difficult to interpret. To simplify serological testing for B. duncani, a proteomics approach was employed to identify candidate immunodiagnostic antigens. Several proteins were identified by electrospray ionization mass spectrometric analysis, and four recombinant protein constructs were expressed and used in a multiplex bead assay (MBA) to detect B. duncani-specific antibodies. Two antigens, AAY83295.1 and AAY83296.1, performed well with high sensitivities and specificities. AAY83295.1 had a higher sensitivity (100%) but lower specificity (89%) than AAY83296.1, which had a sensitivity of 90% and a specificity of 96%. Combining these two antigens did not improve the performance of the assay. This MBA could be useful for diagnosis, serosurveillance, and blood donor screening for B. duncani infection.

Objective To evaluate the diagnostic performance of two commercially available ELISA kits, Novalisa® and Ridascreen® , for the detection of antibodies to Taenia solium, compared to serological diagnosis of neurocysticercosis (NCC) by LLGP-EITB (electro-immunotransfer blot assay using lentil-lectin purified glycoprotein antigens). Methods Archive serum samples from patients with viable NCC (n = 45) or resolved, calcified NCC (n = 45), as well as sera from patients with other cestode parasites (hymenolepiasis, n = 45 and cystic hydatid disease, n = 45), were evaluated for cysticercosis antibody detection using two ELISA kits, Novalisa® and Ridascreen® . All NCC samples had previously tested positive, and all samples from heterologous infections were negative on LLGP-EITB for cysticercosis. Positive rates were calculated by kit and sample group and compared between the two kits. Results Compared to LLGP-EITB, the sensitivity of both ELISA assays to detect specific antibodies in patients with viable NCC was low (44.4% and 22.2%), and for calcified NCC, it was only 6.7% and 4.5%. Sera from patients with cystic hydatid disease were highly cross-reactive in both ELISA assays (38/45, 84.4%; and 25/45, 55.6%). Sera from patients with hymenolepiasis cross-reacted in five cases in one of the assays (11.1%) and in only one sample with the second assay (2.2%). Conclusions The performance of Novalisa® and Ridascreen® was poor. Antibody ELISA detection cannot be recommended for the diagnosis of neurocysticercosis.

Baylisascaris procyonis roundworms can cause potentially fatal neural larva migrans in many species, including humans. However, the clinical spectrum of baylisascariasis is not completely understood. We tested 347 asymptomatic adult wildlife rehabilitators for B. procyonis antibodies; 24 were positive, suggesting that subclinical baylisascariasis is occurring among this population.

Infection with this parasite should be included in differential diagnosis of fever and anemia after blood transfusion or organ transplantation., Babesia microti, an intraerythrocytic parasite, is tickborne in nature. In contrast to transmission by blood transfusion, which has been well documented, transmission associated with solid organ transplantation has not been reported. We describe parasitologically confirmed cases of babesiosis diagnosed ≈8 weeks posttransplantation in 2 recipients of renal allografts from an organ donor who was multiply transfused on the day he died from traumatic injuries. The organ donor and recipients had no identified risk factors for tickborne infection. Antibodies against B. microti parasites were not detected by serologic testing of archived pretransplant specimens. However, 1 of the organ donor’s blood donors was seropositive when tested postdonation and had risk factors for tick exposure. The organ donor probably served as a conduit of Babesia parasites from the seropositive blood donor to both kidney recipients. Babesiosis should be included in the differential diagnosis of unexplained fever and hemolytic anemia after blood transfusion or organ transplantation.

The use of L protein coupled magnetic particles for the concentration and purification of immunoglobulin M (mIgM) monoclonal antibodies against Taenia solium was evaluated. Three concentration methods and different elution times were evaluated and the ratio of particles to the ratio of mIgM was optimized. It is demonstrated that: 1) with the use of magnetic particles, a previous concentration of mIgM is not required, which reduces the manipulation of the antibodies and improves the recovery, 2) the use of a binding buffer can be omitted, since the pH of most cell culture supernatants are neutral, and 3) longer elution times (~ 45 minutes) are needed to increase recovery to a level greater than 80%. The study demonstrates that the use of L protein-coupled magnetic particles is a simple and efficient tool for mIgM concentration and purification.Se evaluó el uso de partículas magnéticas acopladas a proteína L para la concentración y purificación de anticuerpos monoclonales inmunoglobulina M (mIgM) contra Taenia solium. Se evaluaron tres métodos de concentración y diferentes tiempos de elución y se optimizó la proporción de partículas a la proporción de mIgM. Demostramos que: 1) con el uso partículas magnéticas no se requiere de una concentración previa de mIgM, lo que disminuye la manipulación de los anticuerpos y mejora la recuperación, 2) se puede omitir el uso de un tampón de unión, ya que el pH de la mayoría de los sobrenadantes de cultivo celular son neutros, y 3) se necesitan tiempos de elución más largos (~45 minutos) para aumentar la recuperación a un nivel mayor a 80%. El estudio demuestra que el uso de partículas magnéticas acopladas a proteína L es una herramienta simple y eficiente para la concentración y purificación de mIgM.

BACKGROUND Babesiosis is an emerging tick-borne infection in humans. The increasing numbers of reported cases of transfusion-associated babesiosis (TAB), primarily caused by Babesia microti, represents a concern for the safety of the US blood supply. STUDY DESIGN AND METHODS This study investigated kinetics of parasitemia and innate immune responses and dynamics of antibody responses during B. microti infection in rhesus macaques (RMs) using blood smears, quantitative polymerase chain reaction (qPCR), flow cytometry, and indirect fluorescent antibody testing. A total of six monkeys were transfused with either hamster or monkey-passaged B. microti–infected red blood cells (two and four monkeys, respectively) simulating TAB. RESULTS The prepatent period in monkeys inoculated with hamster-passaged B. microti was 35 days compared with 4 days in monkeys transfused with monkey-passaged B. microti; the latter monkeys also had markedly higher parasitemia levels. The duration of the window period from the first detected parasitemia by qPCR analysis to the first detected antibody response ranged from 10 to 17 days. Antibody responses fluctuated during the course of the infection. Innate responses assessed by the frequencies of monocytes and activated B cells correlated with the kinetics and magnitude of parasitemia. On Day 14, additional activation peaks were noted for CD14+CD16+ and CD14–CD16+ monocytes and for CD11c+ myeloid dendritic cells, but only in animals transfused with monkey-passaged B. microti. Parasitemia persisted in these immunocompetent animals, similar to human infection. CONCLUSION The results suggest that transfusion-associated transmission of B. microti leads to rapid onset of parasitemia (Day 4) in RMs, detectable antibody response 14 days later, and persistent parasitemia.

Baylisascaris procyonis (raccoon roundworm) infection is common in raccoons and can cause devastating pathology in other animals, including humans. Limited information is available on the frequency of asymptomatic human infection. We tested 150 adults from California, USA, for B. procyonis antibodies; 11 were seropositive, suggesting that subclinical infection does occur.

In Africa, urbanization is happening faster than ever before which results in new implications for transmission of infectious diseases. For the zoonotic parasite Taenia solium, a major cause of acquired epilepsy in endemic countries, the prevalence in urban settings is unknown. The present study investigated epidemiological, neurological, and radiological characteristics of T. solium cysticercosis and taeniasis (TSCT) in people with epilepsy (PWE) living in Dar es Salaam, Tanzania, one of the fastest growing cities worldwide. A total of 302 PWE were recruited from six health centers in the Kinondoni district of Dar es Salaam. Serological testing for T. solium cysticercosis-antigen (Ag) and -antibodies (Abs) and for T. solium taeniasis-Abs was performed in all PWE. In addition, clinical and radiological examinations that included cranial computed tomography (CT) were performed. With questionnaires, demographic data from study populations were collected, and factors associated with TSCT were assessed. Follow-up examinations were conducted in PWE with TSCT. T. solium cysticercosis-Ag was detected in three (0.99%; 95% CI: 0–2.11%), -Abs in eight (2.65%; 95% CI: 0.84–4.46%), and taeniasis-Abs in five (1.66%; 95% CI: 0.22–3.09%) of 302 PWE. Six PWE (1.99%; 95% CI: 0.41–3.56%) were diagnosed with neurocysticercosis (NCC). This study demonstrates the presence of TSCT in Dar es Salaam, however, NCC was only associated with a few cases of epilepsy. The small fraction of PWE with cysticercosis- and taeniasis-Abs may suggest that active transmission of T. solium plays only a minor role in Dar es Salaam. A sufficiently powered risk analysis was hampered by the small number of PWE with TSCT; therefore, further studies are required to determine the exact routes of infection and risk behavior of affected individuals., Author summary Taenia solium cysticercosis and taeniasis is a zoonotic disease complex which affects thousands of people in sub-Saharan Africa. This parasite has a human-pig life cycle and has been considered a public health problem mainly in rural areas. As African towns and suburbs grow rapidly and disproportionally, adequate infrastructure such as sewage systems and clean water often lack while population density, trade, and travel increase. This may lead to the appearance of parasitic diseases formerly considered `rural´ in urban settings. In this study, we searched for evidence of T. solium infections in the Kinondoni district of Dar es Salaam, Tanzania. We focused on people with epilepsy (PWE) since epilepsy is one of the most common and severe disorders associated with T. solium neurocysticercosis and tested all of them serologically for T. solium cysticercosis and taeniasis. We further investigated neurological and radiological characteristics. Our findings show that in our study area in Dar es Salaam 2.65% of PWE had contracted T. solium infection at some stage. Neurocysticercosis, as confirmed by neuroimaging, was found only in 1.99% of PWE. This, in combination with the relatively small number of PWE detected with taeniasis antibodies (1.66%), points towards the fact that active transmission of T. solium seems to play only a minor role in this urban setting, suggesting that infections may mainly be contracted in rural areas. Further large-scale studies are required to investigate the infection pathways and risk behavior related to T. solium infections within urban areas of sub-Saharan Africa.

Objectives To identify immunodominant antigens of Toxocara canis recognised by Toxocara-infected sera as recombinant reagents for immunodiagnosis of toxocariasis. Methods Pooled sera from human cases of toxocariasis were used to identify immunodominant antigens by immunoscreening a T. canis larval expression cDNA library. The positive clones were sequenced to reveal the identity of the antigens. The recombinant proteins were expressed in E. coli and then used to confirm their immunoreaction with sera of humans with toxocariasis. Two chosen antigens were also used to differentiate Toxocara infection from other helminth infections in mice. Results Eleven antigens with immunodiagnostic potential were identified, including two C-type lectins (CTLs) that reacted strongly with the Toxocara-positive serum pool. The first CTL (Tc-CTL-1) is the same as TES-32, previously identified as a major immunodominant component of TES; the second CTL (Tc-CTL-2) is a novel C-type lectin sharing 83% amino acid sequence identity within the functional domain of Tc-CTL-1. The E. coli-expressed recombinant Tc-CTL-1 was strongly recognised by the Toxocara-positive serum pool or sera from animals experimentally infected with T. canis. Reactivity with recombinant Tc-CTL-1 was higher when the unreduced protein was used in an enzyme-linked immunosorbent assay (ELISA), dot-blot assay or Western blot test compared to the protein under reduced condition. Both recombinant Tc-CTL-1- and Tc-CTL-2-based ELISAs were able to differentiate T. canis infection from other helminth infections in experimentally infected mice. Conclusions Both Tc-CTL-1 and Tc-CTL-2 were able to differentiate Toxocara infection from other helminth infections and could potentially be used as sensitive and specific immunodiagnostic antigens. Objectifs Identifier des antigenes immunodominants de Toxocara canis reconnus par des serums infectes par Toxocara, comme recombinants reactifs pour le diagnostic immunologique de la toxocarose. Methodes Des serums pooles provenant de cas humains de toxocarose ont ete utilises pour identifier des antigenes immunodominants par immunocriblage d'une banque d'expression d’ADNc de larves de T. canis. Les clones positifs ont ete sequences pour reveler l'identite des antigenes. Les proteines recombinantes ont ete exprimees dans E. coli et ensuite utilisees pour confirmer leur reaction immunologique avec des serums humains de toxocarose. Deux antigenes ont egalement ete utilises pour differencier la toxocarose d'autres helminthiases chez la souris. Resultats 11 antigenes a potentiel d'immunodiagnostic ont ete identifies, dont deux lectines de type C (CTL) qui ont reagi fortement avec le pool de serums positifs pour Toxocara. La premiere CTL (Tc-CTL-1) est la meme que TES-32, precedemment identifiee comme une composante immunodominante majeure de TES; la deuxieme CTL (Tc-CTL-2) est une nouvelle lectine de type C partageant une identite de 83% avec la sequence d'acides amines dans la region fonctionnelle de Tc-CTL-1. Le recombinant Tc-CTL-1 exprime dans E. coli a ete fortement reconnu par le pool de serums positifs pour Toxocara ou des serums provenant d'animaux infectes experimentalement avec T. canis. La reactivite avec le recombinant Tc-CTL-1 etait plus forte lorsque la proteine non reduite a ete utilisee dans des essais ELISA, Dot-blot ou Western-blot par rapport a la proteine sous sa forme reduite. Les ELISA bases sur les deux recombinants Tc-CTL-1 et Tc-CTL-2 ont ete en mesure de differencier l'infection a T. canis d'autres helminthiases chez la souris infectee experimentalement. Conclusions Tc-CTL-1 et Tc-CTL-2 ont ete en mesure de differencier la toxocarose d'autres helminthiases et pourraient potentiellement etre utilises comme antigenes d'immunodiagnostic sensibles et specifiques. Objetivos Identificar antigenos inmunodominantes de Toxocara canis reconocidos por sueros infectados con Toxocara como reactivos recombinantes para el inmunodiagnostico de la toxocariasis. Metodos Se utilizo un pool de sueros de casos de toxocariasis humana para identificar los antigenos inmunodominantes mediante inmunoscreening en una biblioteca de ADNc de larvas de T. canis. Los clones que dieron positivo fueron secuenciados para identificar los antigenos. Las proteinas recombinantes se expresaron en E. coli y se utilizaron para confirmar su inmunoreactividad con sueros de humanos con toxocariasis. Dos antigenos especificos se utilizaron tambien para diferenciar la infeccion por Toxocara de otras infecciones por helmintos en raton. Resultados Se identificaron 11 antigenos con potencial inmunodiagnostico, incluyendo dos lectinas del tipo C (CLT) que reaccionaban fuertemente con el pool de sueros Toxocara-positivos. La primera LTC (Tc- CLT -1) es la misma que TES-32, previamente identificada como el principal componente inmunodominante de TES; la segunda LTC (Tc- CLT -2) es una nueva lectina tipo C que comparte un 83% de la secuencia de aminoacidos dentro del dominio funcional de Tc- CLT -1. La Tc- CLT -1 recombinante expresada en E. coli era altamente reconocida por el pool de sueros Toxocara-positivos o sueros de animales infectados experimentalmente con T. canis. La reactividad con el Tc- CLT -1 recombinante era mayor cuando se utilizaba la proteina bajo condiciones no reductoras en un ELISA, Dot blot o Western blot comparado con la proteina en condiciones reductoras. Las ELISAs en las que se utilizaban la Tc- CLT -1 recombinante y la Tc- CLT -2 recombinante eran capaces de diferenciar la infeccion por T. canis de otras infecciones helminticas en ratones infectados experimentalmente. Conclusiones Tanto Tc- CLT-1 como Tc- CLT-2 eran capaces de diferenciar la infeccion por Toxocara de otras infecciones helminticas y potencialmente podrian utilizarse como antigenos sensibles y especificos para inmunodiagnostico.

Despite the global distribution and public health consequences of Taenia tapeworms, the life cycles of taeniids infecting wildlife hosts remain largely undescribed. The larval stage of Taenia serialis commonly parasitizes rodents and lagomorphs, but has been reported in a wide range of hosts that includes geladas (Theropithecus gelada), primates endemic to Ethiopia. Geladas exhibit protuberant larval cysts indicative of advanced T. serialis infection that are associated with high mortality. However, non-protuberant larvae can develop in deep tissue or the abdominal cavity, leading to underestimates of prevalence based solely on observable cysts. We adapted a non-invasive monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) to detect circulating Taenia spp. antigen in dried gelada urine. Analysis revealed that this assay was highly accurate in detecting Taenia antigen, with 98.4% specificity, 98.5% sensitivity, and an area under the curve of 0.99. We used this assay to investigate the prevalence of T. serialis infection in a wild gelada population, finding that infection is substantially more widespread than the occurrence of visible T. serialis cysts (16.4% tested positive at least once, while only 6% of the same population exhibited cysts). We examined whether age or sex predicted T. serialis infection as indicated by external cysts and antigen presence. Contrary to the female-bias observed in many Taenia-host systems, we found no significant sex bias in either cyst presence or antigen presence. Age, on the other hand, predicted cyst presence (older individuals were more likely to show cysts) but not antigen presence. We interpret this finding to indicate that T. serialis may infect individuals early in life but only result in visible disease later in life. This is the first application of an antigen ELISA to the study of larval Taenia infection in wildlife, opening the doors to the identification and description of infection dynamics in reservoir populations., Author summary Although tapeworm parasites of the genus Taenia are globally distributed and inflict enormous socioeconomic and health costs on their hosts, which include humans, little is known about taeniid tapeworms that infect wildlife. This gap in knowledge prevents an assessment of the potential for these parasites to infect humans and production animals and is largely due to the difficulty of conducting standard diagnostic tests on wildlife. To address this gap, we adapted a standard diagnostic assay to be used with dried urine samples. We used urine from geladas, primates endemic to Ethiopia, which are frequently infected with the larval stage of a taeniid tapeworm and exhibit protuberant cysts during advanced infection. The use of this diagnostic test in a wild gelada population allowed us to detect that individuals can be infected without exhibiting observable cysts, and that some individuals may control infection in its early stages. This tool provides information about how a neglected tapeworm functions in a wildlife system and opens the door to the non-invasive identification of tapeworm reservoir hosts that may threaten humans.

Cysticercosis is a potentially fatal and preventable neglected parasitic infection caused by the larval form of Taenia solium. Patients with symptomatic disease usually have signs and symptoms of neurocysticercosis, which commonly manifest as seizures or increased intracranial pressure. Although there are many persons living in the United States who emigrated from highly disease-endemic countries and there are foci of autochthonous transmission of the parasite in the United States, little is known about burden and epidemiology of the disease in this country. In addition, despite advances in the diagnosis and management of neurocysticercosis, there remain many unanswered questions. Improving our understanding and management of neurocysticercosis in the United States will require improved surveillance or focused prospective studies in appropriate areas and allocation of resources towards answering some of the key questions discussed in this report.

Schistosomiasis is a debilitating parasitic disease of humans, endemic in tropical areas, for which no vaccine is available. Evidence points to glycan antigens as being important in immune responses to infection. Here we describe our studies on the comparative humoral immune responses to defined schistosome-type glycan epitopes in Schistosoma mansoni-infected humans, rhesus monkeys and mice. Rhesus anti-glycan responses over the course of infection were screened on a defined glycan microarray comprising semi-synthetic glycopeptides terminating with schistosome-associated or control mammalian-type glycan epitopes, as well as a defined glycan microarray of mammalian-type glycans representing over 400 glycan structures. Infected rhesus monkeys generated a high immunoglobulin G (IgG) antibody response to the core xylose/core α3 fucose epitope of N-glycans, which peaked at 8–11 weeks post infection, coinciding with maximal ability to kill schistosomula in vitro. By contrast, infected humans generated low antibody levels to this epitope. At 18 months following praziquantel therapy to eliminate the parasite, antibody levels were negligible. Mice chronically infected with S. mansoni generated high levels of anti-fucosylated LacdiNAc (GalNAcβ1, 4(Fucα1, 3)GlcNAc) IgM antibodies, but lacked a robust response to the core xylose/core α3 fucose N-glycan antigens compared with other species studied, and their sera demonstrated an intermediate level of schistosomula killing in vitro. These differential responses to parasite glycan antigens may be related to the ability of rhesus monkeys to self-cure in contrast to the chronic infection seen in humans and mice. Our results validate defined glycan microarrays as a useful technology to evaluate diagnostic and vaccine antigens for schistosomiasis and perhaps other infections.

The clinical spectrum of human disease caused by Toxocara canis and Toxocara cati ranges from visceral and ocular larva migrans to covert and common toxocariasis. Since the parasite is not typically recovered in affected tissues, detection of parasite-specific antibodies is an important and necessary step in establishing a diagnosis of toxocariasis. Most immunodiagnostic methods have historically used and continue to use the Toxocara excretory-secretory antigens (TES-Ag) in an ELISA format to detect Toxocara-specific antibodies. The TES-Ag ELISA has proven to be specific, robust, and reliable, although questions about specificity and reduced sensitivity leave ample room for improvement in laboratory diagnosis of toxocariasis. Recent advances have focused on development of recombinant protein targets to measure Toxocara-specific antibodies. The potential for development of new diagnostics using recombinant proteins is presented herein, as well as possible roles for antigen and molecular detection methods.

SUMMARYThe primary causative agent of eosinophilic meningoencephalitis (EoM) in endemic regions is the nematode Angiostrongylus cantonensis. The occurrence of EoM was previously restricted to countries in Southeast Asia and the Pacific Islands; however, more recently, it has been reported from other regions, including Brazil. The commonly used diagnosis is detection of specific antibody reactivity to the 31 kDa antigen, which is derived from female worm somatic extracts. Here we report the occurrence of cross-reactivity to this antigen in sera from other parasitic infections, especially those that may cause EoM, such as gnathostomiasis, toxocariasis, hydatidosis and strongyloidiasis. We also demonstrated that the cross-reactivity, in part, is dependent of the concentration of antigen used in Western blot assays. We discuss the importance of these findings on the interpretation of this test.

We tested refugee camp residents on the Thailand–Myanmar border for Taenia solium infection. Taeniasis prevalence was consistent with that for other disease-endemic regions, but seropositivity indicating T. solium taeniasis was rare. Seropositivity indicating cysticercosis was 5.5% in humans, and 3.2% in pigs. Corralling pigs and providing latrines may control transmission of these tapeworms within this camp.

Purpose The prevalence of epilepsy in Cameroon is higher than that of the industrialized world and other developing countries. Neurocysticercosis due to Taenia solium infestation has been reported as a major cause of epilepsy in some parts of Cameroon although there are some conflicting data. The prevalence of epilepsy is especially high in the Momo division of the North-West Province of Cameroon. We hypothesized that individuals with epilepsy in this region have a higher percentage of seropositivity to T. solium than matched controls. Methods We conducted a case–control study in the Momo subdivision of Ngie. Individuals with epilepsy were recruited from the health centers in Ngie. Control subjects were selected from 19 Ngie villages. Potential cases of people with epilepsy (PWE) were identified through a questionnaire applied by trained field workers, using history of epileptic seizures as a key indicator. Blood samples were taken from all consenting individuals by finger prick, stored in StabilZyme Select, and assayed for antibodies to T. solium in an Atlanta based reference laboratory. Results We accrued 249 patients with epilepsy, of whom 237 met the inclusion criteria, and 245 age-matched controls. There was no significant difference in seropositivity to T. solium between those individuals with epilepsy (5%) and controls (4.9%). Conclusions Our data do not support the hypothesis that epilepsy is associated with seropositivity to T. solium . It is highly unlikely that cysticercosis plays a causative role in the high prevalence of epilepsy in this region of Cameroon.

Infection of mammals by the parasitic helminth Schistosoma mansoni induces antibodies to glycan antigens in worms and eggs, but the differential nature of the immune response among infected mammals is poorly understood. To better define these responses, we used a shotgun glycomics approach in which N-glycans from schistosome egg glycoproteins were prepared, derivatized, separated, and used to generate an egg shotgun glycan microarray. This array was interrogated with sera from infected mice, rhesus monkeys, and humans and with glycan-binding proteins and antibodies to gather information about the structures of antigenic glycans, which also were analyzed by mass spectrometry. A major glycan antigen targeted by IgG from different infected species is the FLDNF epitope [Fucα3GalNAcβ4(Fucα3)GlcNAc-R], which is also recognized by the IgG monoclonal antibody F2D2. The FLDNF antigen is expressed by all life stages of the parasite in mammalian hosts, and F2D2 can kill schistosomula in vitro in a complement-dependent manner. Different antisera also recognized other glycan determinants, including core β-xylose and highly fucosylated glycans. Thus, the natural shotgun glycan microarray of schistosome eggs is useful in identifying antigenic glycans and in developing new anti-glycan reagents that may have diagnostic applications and contribute to developing new vaccines against schistosomiasis.

Background The frequency of Taenia solium, a zoonotic helminth, is increasing in many countries of sub-Saharan Africa, where the prevalence of the human immunodeficiency virus (HIV) is also high. However, little is known about how these two infections interact. The aim of this study was to compare the proportion of HIV positive (+) and negative (−) individuals who are infected with Taenia solium (TSOL) and who present with clinical and neurological manifestations of cysticercosis (CC). Methods In northern Tanzania, 170 HIV+ individuals and 170 HIV– controls matched for gender, age and village of origin were recruited. HIV staging and serological tests for TSOL antibodies (Ab) and antigen (Ag) were performed. Neurocysticercosis (NCC) was determined by computed tomography (CT) using standard diagnostic criteria. Neurological manifestations were confirmed by a standard neurological examination. In addition, demographic, clinical and neuroimaging data were collected. Further, CD4+ cell counts as well as information on highly active antiretroviral treatment (HAART) were noted. Results No significant differences between HIV+ and HIV– individuals regarding the sero-prevalence of taeniosis-Ab (0.6% vs 1.2%), CC-Ab (2.4% vs 2.4%) and CC-Ag (0.6% vs 0.0%) were detected. A total of six NCC cases (3 HIV+ and 3 HIV–) were detected in the group of matched participants. Two individuals (1 HIV+ and 1 HIV–) presented with headaches as the main symptom for NCC, and four with asymptomatic NCC. Among the HIV+ group, TSOL was not associated with CD4+ cell counts, HAART duration or HIV stage. Conclusions This study found lower prevalence of taeniosis, CC and NCC than had been reported in the region to date. This low level of infection may have resulted in an inability to find cross-sectional associations between HIV status and TSOL infection or NCC. Larger sample sizes will be required in future studies conducted in that area to conclude if HIV influences the way NCC manifests itself.

We communicate a case of a middle-aged Brazilian patient with an unusual presentation of fourth ventricular neurocysticercosis: occurrence of two intraventricular cysts at different locations in the brain within 2 years and cognitive decline as the only neurological symptom. Neurocysticercosis was confirmed by magnetic resonance imaging, serology, histology, and genetic analysis. Neurocysticercosis should be considered as a differential diagnosis in cases with atypical neurologic or psychiatric symptoms, atypical neuroimaging, and travel history. Especially, fourth ventricular cysts carry the risk of obstructive hydrocephalus and brainstem compression and therefore should be extirpated completely. If complete removal of the cystic structures cannot be proven in cases with surgically treated neurocysticercosis, anthelminthic therapy and thorough follow-up examinations should be conducted.

In October of 2008, an outbreak of trichinellosis occurred in northern California that sickened 30 of 38 attendees of an event at which meat from a black bear was served. Morphologic and molecular testing of muscle from the leftover portion of bear meat revealed that the bear was infected with Trichinella murrelli, a sylvatic species of Trichinella found in temperate North America. Clinical records revealed a high attack rate for this outbreak: 78% for persons consuming any bear meat and 100% for persons consuming raw or undercooked bear meat. To our knowledge, this report is the first published report of a human trichinellosis outbreak in the United States attributed to T. murrelli, and it is the second such outbreak reported worldwide.

Cysticercosis is an infection caused by a pork tapeworm that creates cysts in different areas of the human body. Sometimes, these parasites can get into the infected patient’s brain and lead to epilepsy or other neurologic disorders. Cysticercosis is most common in developing countries that have poor sanitation and where pigs feed on human waste; however, cases in the United States are increasing. A recent study found that many refugees who settle in the United States, including those from Burma, Laos, Burundi, and Bhutan, have been infected with the tapeworm. The occurrence of cysticercosis among these groups has clinical and public health implications because US physicians might not be familiar with this disease and its symptoms. Cysticercosis should be suspected in refugees who have seizures, headache, or other unexplained neurologic symptoms. Physicians should also be aware that treatment for intestinal parasites, routinely given to refugees before they leave their homeland, can cause serious neurologic reactions in those already infected with the tapeworm., Neurocysticercosis (NCC) is a disease caused by central nervous system infection by the larval stage of the pork tapeworm, Taenia solium. In developing countries, NCC is a leading cause of adult-onset epilepsy. Case reports of NCC are increasing among refugees resettled to the United States and other nations, but the underlying prevalence among refugee groups is unknown. We tested stored serum samples from the Centers for Disease Control and Prevention Migrant Serum Bank for antibodies against T. solium cysts by using the enzyme-linked immunoelectrotransfer blot. Seroprevalence was high among all 4 populations tested: refugees from Burma (23.2%), Lao People’s Democratic Republic (18.3%), Bhutan (22.8%), and Burundi (25.8%). Clinicians caring for refugee populations should suspect NCC in patients with seizure, chronic headache, or unexplained neurologic manifestations. Improved understanding of the prevalence of epilepsy and other associated diseases among refugees could guide recommendations for their evaluation and treatment before, during, and after resettlement.

Anthrax is endemic throughout Africa, causing considerable livestock and wildlife losses and severe, sometimes fatal, infection in humans. Predicting the risk of infection is therefore important for public health, wildlife conservation and livestock economies. However, because of the intermittent and variable nature of anthrax outbreaks, associated environmental and climatic conditions, and diversity of species affected, the ecology of this multihost pathogen is poorly understood. We explored records of anthrax from the Serengeti ecosystem in north-west Tanzania where the disease has been documented in humans, domestic animals and a range of wildlife. Using spatial and temporal case-detection and seroprevalence data from wild and domestic animals, we investigated spatial, environmental, climatic and species-specific associations in exposure and disease. Anthrax was detected annually in numerous species, but large outbreaks were spatially localized, mostly affecting a few focal herbivores. Soil alkalinity and cumulative weather extremes were identified as useful spatial and temporal predictors of exposure and infection risk, and for triggering the onset of large outbreaks. Interacting ecological and behavioural factors, specifically functional groups and spatiotemporal overlap, helped to explain the variable patterns of infection and exposure among species. Synthesis and applications. Our results shed light on ecological drivers of anthrax infection and suggest that soil alkalinity and prolonged droughts or rains are useful predictors of disease occurrence that could guide risk-based surveillance. These insights should inform strategies for managing anthrax including prophylactic livestock vaccination, timing of public health warnings and antibiotic provision in high-risk areas. However, this research highlights the need for greater surveillance (environmental, serological and case-detection-orientated) to determine the mechanisms underlying anthrax dynamics. Keywords: Bacillus anthracis, disease ecology, exposure, infectious disease, multihost, serology, surveillance, susceptibility, zoonosis Introduction Bacillus anthracis (Cohn), the causative agent of anthrax, is a multihost pathogen affecting human, livestock and wildlife populations. The disease has a world-wide distribution, but has declined in many developed countries because of the implementation of livestock vaccination programmes and sanitary measures. However, anthrax is still endemic in Africa, with severe outbreaks causing significant losses in domestic and wild animal populations (Prins & Weyerhaeuser 1987; Shiferaw et al. 2002; Siamudaala 2006; Clegg et al. 2007; Wafula, Patrick & Charles 2008). Non-fatal cutaneous anthrax acquired through contact with infected carcasses or animal products accounts for >95% of all reported human cases globally (World Health Organization 2008). More severe forms, particularly gastrointestinal anthrax because of handling and consumption of inadequately cooked products from infected animals, may also exert a substantial burden that is both underreported and under-diagnosed (Sirisanthana & Brown 2002). Predicting the risk of infection is therefore important from the perspectives of public health, wildlife conservation and livestock economies. Despite the infamous reputation of anthrax, the ecology and transmission of the disease under natural conditions are not well understood (Hugh-Jones & de Vos 2002). The difficulty lies in the intermittent and variable nature of outbreaks, with considerable variation in the species affected, and associated environmental and climatic conditions (Table 1). Table 1 Anthrax outbreaks in Eastern and Southern Africa for which details were available. Predominant species are emboldened, followed by species in which > 5 cases were documented The life cycle of B. anthracis comprises a multiplication phase in the mammalian host and a persistence phase of spores in the soil. Transmission to herbivores largely occurs indirectly, so risk factors are usually associated with exposure to spores, rather than direct animal-to-animal transmission. Survival mechanisms of the pathogen outside the host remain unclear. Bacillus anthracis has been shown to be associated with plant roots (Saile & Koehler 2006), and this is suggested to be an adaptation that increases the likelihood of infecting ungulate hosts (Raymond et al. 2010). Certain environmental factors affect long-term spore survival and thus increase the risk of anthrax (Blackburn et al. 2007). For example, the endemicity of B. anthracis in some areas has been associated with calcium-rich and neutral-to-alkaline soils (van Ness 1971; Dragon et al. 2005; Hugh-Jones & Blackburn 2009), although strain differences exist in soil chemistry preferences (Smith et al. 2000). Strain differences, in fact, may also govern spread in anthrax epidemics (Blackburn et al. 2007; Garofolo et al. 2010). Seasonal incidence patterns suggest climatic factors (precipitation and ambient temperature) play an important role in triggering outbreaks, although these are not consistent between locations (Table 1), and therefore underlying mechanisms are debated. In some African ecosystems, outbreaks have been reported late in the dry season (Prins & Weyerhaeuser 1987; Turnbull et al. 1991; Lindeque & Turnbull 1994; de Vos & Bryden 1996; Shiferaw et al. 2002; Clegg et al. 2007; Muoria et al. 2007), leading to suggestions that transmission may be facilitated by close grazing, nutritional stress and congregation at water holes (Hugh-Jones & de Vos 2002). Elsewhere, outbreaks have been associated with heavy rains (Lindeque & Turnbull 1994; Mlengeya et al. 1998; Wafula, Patrick & Charles 2008; Bellan 2010), which are hypothesized to unearth spores and amplify vector populations (Durrheim et al. 2009; Hugh-Jones & Blackburn 2009; Lewerin et al. 2010). Experimental spore germination and vegetative growth in the rhizosphere provide a mechanism by which bursts of rainfall could spark epidemics (Saile & Koehler 2006). This would parsimoniously explain anthrax occurrences during dry summers in North America and Australia that have been preceded by rains and when animals are often in good body condition (Turner et al. 1999; Hugh-Jones & de Vos 2002; Parkinson, Rajic & Jenson 2003; Hugh-Jones & Blackburn 2009; Epp, Waldner & Argue 2010). Although anthrax infects a wide range of species, outbreaks are typically associated with just a few (Table 1), for example impala Aepyceros melampus, kudu Tragelaphus strepsiceros, buffalo Syncerus caffer and zebra Equus spp. (Prins & Weyerhaeuser 1987; Mlengeya et al. 1998; Clegg et al. 2007; Muoria et al. 2007; Wafula, Patrick & Charles 2008). Even within a single ecosystem, different species may be affected at different times (Lindeque & Turnbull 1994). Reasons for this remain unclear, but could be attributed to host-specific differences in susceptibility and exposure as a result of behavioural and ecological traits as well as differences in pathogen strains. The overall ecological and genetic factors that contribute to environmental persistence of anthrax, and the underlying conditions that initiate outbreaks and underlie patterns of circulation are poorly known. In the Serengeti ecosystem, anthrax has been reported in a variety of species, with sporadic outbreaks occurring in relatively localized endemic foci and mostly affecting a few focal species (Lembo et al. 2011). The ecosystem covers a large area (over 20 000 km2, Sinclair et al. 2008) encompassing a variety of habitats and environmental gradients, which may influence anthrax occurrence. Here, we draw on data gathered opportunistically including serology indicative of exposure and case reports from human, domestic and wild animal populations in the greater Serengeti ecosystem, to explore ecological factors affecting infection patterns in a range of species. Combined, the investigation into environmental and climatic predictors of anthrax and their associations with species-specific patterns of exposure and mortality provides a holistic picture of ecological drivers of anthrax dynamics and identifies priorities for further research in anthrax endemic ecosystems.

The free-living amoebaBalamuthia mandrillariscauses granulomatous amoebic encephalitis (GAE) in humans. Rapid identification of balamuthiasis is critical for effective therapeutic intervention and case management. In the present study we identified target antigens for the development of a serological assay forB. mandrillarisinfection. We demonstrated by silver staining that protein profiles for all eight isolates ofB. mandrillaris, independent of human or animal origin or geographic origin, appeared to be similar except for some minor differences, indicating the molecular homogeneity of these strains. The profiles of all isolates, which ranged from 200 to 10 kDa, were similar, with a prominent protein visible around 30 kDa; all appeared considerably different from protein profiles of the control E6 cells andAcanthamoeba castellaniiandNaegleria fowleriisolates. Western blot analysis with rabbit hyperimmune serum identified the major immunodominant antigens of 25, 50, 75, and 80 kDa; positive human sera reacted strongly with proteins around 25, 40, 50, and 75 kDa. Proteins around 40 kDa detected by human serum were not recognized by hyperimmune rabbit serum. None of the target proteins were detected by uninfected control sera. Reactivities of five patients' sera with 4 different isolates ofB. mandrillaris(2 strains of human and 2 strains of animal origins) revealed that patients' sera reacted slightly differently with differentB. mandrillarisisolates, although major proteins of approximately 25, 50, and 75 kDa were present in all extracts.

Bacillus anthracis, the bacterium that causes anthrax, is responsible for varying death rates among animal species. Difficulties in case detection, hazardous or inaccessible carcasses, and misdiagnosis hinder surveillance. Using case reports and a new serologic assay that enables multispecies comparisons, we examined exposure to and illness caused by B. anthracis in different species in the Serengeti ecosystem in Tanzania during 1996-2009 and the utility of serosurveillance. High seroprevalence among carnivores suggested regular nonfatal exposure. Seropositive wildebeest and buffalo showed that infection was not invariably fatal among herbivores, whereas absence of seropositivity in zebras and frequent detection of fatal cases indicated high susceptibility. Exposure patterns in dogs reflected known patterns of endemicity and provided new information about anthrax in the ecosystem, which indicated the potential of dogs as indicator species. Serosurveillance is a valuable tool for monitoring and detecting anthrax and may shed light on mechanisms responsible for species-specific variability in exposure, susceptibility, and mortality rates.

Taeniasis/cysticercosis caused by Taenia solium is a frequent parasitic infection of the human brain in most of the world. Rapid and simple screening tools to identify taeniasis and cysticercosis cases are needed for control programs, mostly to identify tapeworm carriers which are the source of infection and need to be treated, or as tools for point-of-care case detection or confirmation. These screening assays should be affordable, reliable, rapid, and easy to perform. Immunochromatographic tests meet these criteria. To demonstrate proof of principle, we developed and evaluated two magnetic immunochromatographic tests (MICTs) for detection of human Taenia solium taeniasis antibodies (ES33-MICT) and neurocysticercosis antibodies (T24-MICT). These assays detected stage-specific antibodies by using two recombinant proteins, rES33 for detection of taeniasis antibodies and rT24H for detection of cysticercosis antibodies. The sensitivity and specificity of the ES33-MICT to detect taeniasis infections were 94.5% and 96%, respectively, and those of the T24-MICT to detect cases of human cysticercosis with two or more viable brain cysts were 93.9% and 98.9%, respectively. These data provide proof of principle that the ES33- and T24-MICTs provide rapid and suitable methods to identify individuals with taeniasis and cysticercosis.

One of the best-characterized tests for the diagnosis of neurocysticercosis is the enzyme-linked immunoelectrotransfer blot assay, developed at the CDC, which uses lentil lectin-purified glycoproteins (LLGPs) extracted from Taenia solium cysticerci. The purification of the LLGP antigens has been difficult to standardize, and the polyacrylamide gel system used for the immunoblot assay is not easily transferable to other laboratories. In this study, we developed a multiantigen printing immunoassay (MAPIA) to compare the performance of multiple recombinant Taenia solium proteins with the potential for the detection of cysticercosis and taeniasis. We prepared MAPIA strips using six cysticercosis and two taeniasis diagnostic proteins and compared the performance of the proteins with sera collected from defined cysticercosis and taeniasis cases. Of the six cysticercosis antigens, rT24H performed well in detecting cases with two or more viable cysts in the brain (sensitivity and specificity, 97% and 99.4%, respectively); the use of a combination of cysticercosis antigens did not improve the sensitivity of the test and decreased the specificity. None of the antigens could differentiate the different clinical presentations of cysticercosis. Both of the taeniasis antigens (rES33 and rES38) had the same sensitivity of 99.4% and specificities of 93.9% and 94.5%, respectively. Some crossreactivity against rES33 and rES38 was found, especially with sera from cases infected with Schistosoma mansoni. We conclude that MAPIA is a simple and effective tool that may be used to compare antibody responses to different cysticercosis and taeniasis antigens and, in this case, may be useful for the rapid detection of T. solium cases. Excellent laboratory methods with high specificities and sensitivities for the immunodiagnosis of neurocysticercosis and taeniasis exist. The enzyme immunoelectrotransfer blot (EITB) for cysticercosis is accepted as the “gold standard” assay for the serological identification of cysticercosis (16, 19). Unfortunately, the test employs complex native proteins in immunoblot assay formats, and therefore, the tests are not easily adaptable to field use. Over the last 10 years we systematically purified and cloned the diagnostic glycoproteins expressed in the lentil lectin glycoprotein fraction. We found that the seven diagnostic proteins are members of three antigenic protein families: the GP50, GP24, and 8-kDa families. The recombinant proteins or synthetic peptides identified in the first-generation assays are available for further comparative analysis.

Currently there are no commercially available selective media indicated for the isolation of Burkholderia mallei and Burkholderia pseudomallei. Ashdown's agar, a custom selective medium for isolation of B. pseudomallei, is well described in the literature but unavailable commercially. Three commercially available media, Burkholderia cepacia selective agar (BCSA), oxidative-fermentative-polymyxin B-bacitracin-lactose (OFPBL) agar, and Pseudomonas cepacia (PC) agar are recommended for isolation of B. cepacia from respiratory secretions of cystic fibrosis patients. We evaluated the sensitivity and selectivity of these four media using 20 B. mallei, 20 B. pseudomallei, 20 Burkholderia spp., and 15 diagnostically challenging organisms. Ashdown's agar was the most sensitive medium for the isolation of B. pseudomallei, but it was unable to support growth of B. mallei. Pseudomonas cepacia agar was highly sensitive and selective for both organisms. In non-endemic areas, we suggest the use of the commercially available PC agar for the isolation of B. mallei and B. pseudomallei.

Centers for Disease Control guidelines for schistosomiasis and strongyloidiasis in Sudanese and Somali refugees are not widely implemented. Given limited prevalence data, we conducted a seroprevalence study of schistosomiasis, strongyloidiasis, and loiasis in Sudanese refugees across diverse ages. Sudanese refugees, ages 4-78, were recruited via community organizations. Half of the patients (86/172), were seropositive for schistosomiasis (46/171; 26.9%), strongyloidiasis (56/172; 33%), or both (16/171; 9.4%). No Loa loa infections were detected. Infection rates were similar in adults and children except that no schistosomiasis was detected in children < 4 years of age at the time of immigration to the United States. The high prevalence of schistosomiasis and strongyloidiasis in a community-based sample of Sudanese confirms the urgency for compliance with CDC refugee health guidelines. We detected no co-infection with Loa loa using the most sensitive serologic techniques, allowing use of ivermectin, the most effective treatment of strongyloidiasis.

We evaluated the effectiveness of an overseas pre-departure regimen of five days of albendazole for presumptive treatment of intestinal parasites by examining stool specimens in treated and untreated Montagnard refugees after arrival in the United States. Among 815 refugees evaluated, fully treated refugees had a significantly lower prevalence of helminths (11 [1.4%] of 777), specifically hookworm and Ascaris lumbricoides, than untreated pregnant women (3 [20%] of 15) (P < 0.001). Multivariate analysis showed that treatment was associated with significantly lower rates of infection with helminths but not protozoa. Post-arrival gastrointestinal symptoms were not associated with findings on stool examination. Our evaluation suggests that although additional studies are needed to determine optimal treatment regimens for intestinal parasites, especially among young children and pregnant women, a five-day course of pre-departure albendazole was effective in reducing helminthic infection in treated refugees.

This report describes the first case of naturally acquired inhalation anthrax in the United States since 1976. The patient's clinical course included adjunctive treatment with human anthrax immunoglobulin. Clinical correlation of serologic assays for the lethal factor component of lethal toxin and anti-protective antigen immunoglobulin G are also presented.

Objective To assess the contribution of neurocysticercosis (NCC) to the burden of epilepsy in a rural Tanzanian population. Methods We identified adult people with epilepsy (PWE) in a door-to-door study in an established demographic surveillance site. PWE and community controls were tested for antibodies to Taenia solium, the causative agent of NCC, and all PWE were offered a computed tomography (CT) head scan. Data on household occupancy and sanitation, pig-keeping and pork consumption were collected from PWE and controls and associations with epilepsy were assessed using chi-square or Fisher's exact tests. Results Six of 218 PWE had antibodies to T. solium (2.8%; 95% CI 0.6–4.9), compared to none of 174 controls (Fisher's exact test, P = 0.04). Lesions compatible with NCC were seen in eight of 200 CT scans (4.0%; 95% CI 1.3–6.7). A total of 176 PWE had both investigations of whom two had positive serology along with NCC-compatible lesions on CT (1.1%; 95% 0.3–4.0). No associations between epilepsy and any risk factors for NCC were identified. Conclusions Neurocysticercosis is present in this population but at a lower prevalence than elsewhere in Tanzania and sub-Saharan Africa. Insights from low-prevalence areas may inform public health interventions designed to reduce the burden of preventable epilepsy. Objectif Evaluer la contribution de la neurocysticercose (NCC) a la charge de l’epilepsie dans une population rurale de la Tanzanie. Methodes Nous avons identifie les personnes adultes souffrant d’epilepsie (PSE) dans une etude de porte-a-porte dans un site etabli de surveillance demographique. Les PSE et des temoins communautaires ont ete testes pour les anticorps de Taenia solium, l'agent causal de la NCC et toutes les PSE ont ete offertes une tomodensitometrie (TDM) de la tete. Les donnees sur l'occupation des menages et l'assainissement, l’elevage et la consommation de porc ont ete recueillies chez les PSE et les temoins et les associations avec l’epilepsie ont ete evaluees en utilisant les tests Exacts de Fisher ou de Chi-carre. Resultats Six des 218 PSE avaient des anticorps de T. Solium (2,8%; IC95%: 0,6 a 4,9) compares a aucun des 174 temoins (test Exact de Fisher, p = 0,04). Les lesions compatibles avec la NCC ont ete observees dans 8 des 200 TDM (4,0%; IC95%: 1,3 a 6,7). Les deux investigations ont ete menees chez 176 PSE, dont deux avaient une serologie positive ainsi que des lesions compatibles avec la NCC sur la TDM (1,1%; IC95%: 0,3 a 4,0). Aucune association entre l’epilepsie et les facteurs de risque categorises pour la NCC n'a ete identifiee. Conclusions La NCC est presente dans cette population, mais a un taux de prevalence plus faible qu'ailleurs en Tanzanie et en Afrique subsaharienne. Les donnees des zones a faible prevalence pourraient eclairer les interventions de sante publique visant a reduire la charge evitable de l’epilepsie. Objetivo Evaluar la contribucion de la neurocisticercosis (NCC) a la carga de epilepsia entre la poblacion rural de Tanzania. Metodos Identificamos a adultos con epilepsia (ACE) en un estudio puerta-a-puerta, en un lugar con vigilancia demografica establecida. A los ACE y a los controles comunitarios se les realizaron pruebas en busca de anticuerpos contra Taenia solium, el agente causal de la NCC, y a todos los ACE se les ofrecio una tomografia computarizada de la cabeza (TC). Se recolectaron los datos sobre ocupacion del hogar y saneamiento, la crianza y consumo del cerdo de los ACE y de los controles, y se evaluaron las asociaciones con la epilepsia utilizando las pruebas del Chi-cuadrado y de Fisher. Resultados Seis de 218 PCE tenian anticuerpos contra T. solium (2.8%; IC 95% 0.6 a 4.9), comparado con ninguno de los 174 controles (prueba de Fisher, p=0.04). Las lesiones compatibles con la NCC se observaron en ocho de 200 TC (4.0%; IC95% 1.3 a 6.7). A 176 ACE se les realizaron ambas pruebas, y de estos, dos tenian una serologia positiva junto con lesiones compatibles con NCC en la TC (1.1%; 95% 0.3 a 4.0). No se identificaron asociaciones entre la epilepsia y factores de riesgo categoricos para NCC. Conclusiones La NCC esta presente en esta poblacion, pero en una menor prevalencia que en cualquier otro lugar en Tanzania y Africa subsahariana. La comprension de lo que sucede en areas de baja prevalencia podria informar intervenciones de salud publica disenadas para reducir la carga de epilepsia prevenible.

The clinical spectrum of human disease caused by the roundworms Toxocara canis and Toxocara cati ranges from visceral and ocular larva migrans to covert toxocariasis. The parasite is not typically recovered in affected tissues, so detection of parasite-specific antibodies is usually necessary for establishing a diagnosis. The most reliable immunodiagnostic methods use the Toxocara excretory-secretory antigens (TES-Ag) in ELISA formats to detect Toxocara-specific antibodies. To eliminate the need for native parasite materials, we identified and purified immunodiagnostic antigens using 2D gel electrophoresis followed by electrospray ionization mass spectrometry. Three predominant immunoreactive proteins were found in the TES; all three had been previously described in the literature: Tc-CTL-1, Tc-TES-26, and Tc-MUC-3. We generated Escherichia coli expressed recombinant proteins for evaluation in Luminex based immunoassays. We were unable to produce a functional assay with the Tc-MUC-3 recombinant protein. Tc-CTL-1 and Tc-TES-26 were successfully coupled and tested using defined serum batteries. The use of both proteins together generated better results than if the proteins were used individually. The sensitivity and specificity of the assay for detecting visceral larval migrans using Tc-CTL-1 plus Tc-TES-26 was 99% and 94%, respectively; the sensitivity for detecting ocular larval migrans was 64%. The combined performance of the new assay was superior to the currently available EIA and could potentially be employed to replace current assays that rely on native TES-Ag., Author Summary The roundworms Toxocara canis and Toxocara cati cause a broad spectrum of clinical disease in humans. Children are at particular risk of toxocariasis when they play in areas potentially contaminated with Toxocara eggs, such as playgrounds or sandboxes and ingest embryonated roundworm eggs. Currently, diagnosis for toxocariasis relies on clinical signs, history of exposure to puppies or kittens, laboratory findings (including eosinophilia), and the detection of antibodies to Toxocara antigens. The enzyme immunoassay using T. canis excretory secretory antigens from infective-stage larvae is the most useful diagnostic test for toxocaral visceral larva migrans (VLM) and ocular larva migrans (OLM) and is the preferred assay used by most laboratories in the U.S. and worldwide. Although the EIA has been robust and reliable, improvement should be made in the specificity of the assay and the availability of a consistent antigen source. The crude TES-Ag shows cross-reactivity with antibodies from other common helminth infections of humans which reduces the usefulness of native, unfractionated TES Ag-based serodiagnosis in regions where poly-parasitism is endemic. To improve the assay performance, target antigenic proteins from T. canis excretory secretory antigens were identified using 2D gel electrophoresis. Three antigenic proteins sequences were found, expressed, and developed into Luminex bead-based assays. The combined use of two recombinant antigens (Tc-CTL-1 and Tc-TES-26) represents an improvement over the existing immunodiagnostic methods that rely on native parasite materials. In the future, additional antigens could be added from other parasites that cause larval migrans to form a single method for detecting larval migrans syndromes.

Bacillus cereus is ubiquitous in nature, and while most isolates appear to be harmless, some are associated with food-borne illnesses, periodontal diseases, and other more serious infections. In one such infection, B. cereus G9241 was identified as the causative agent of a severe pneumonia in a Louisiana welder in 1994. This isolate was found to harbor most of the B. anthracis virulence plasmid pXO1 (13). Here we report the characterization of two clinical and one environmental B. cereus isolate collected during an investigation of two fatal pneumonia cases in Texas metal workers. Molecular subtyping revealed that the two cases were not caused by the same strain. However, one of the three isolates was indistinguishable from B. cereus G9241. PCR analysis demonstrated that both clinical isolates contained B. anthracis pXO1 toxin genes. One clinical isolate and the environmental isolate collected from that victim's worksite contained the cap A , B , and C genes required for capsule biosynthesis in B. anthracis . Both clinical isolates expressed a capsule; however, neither was composed of poly- d -glutamic acid. Although most B. cereus isolates are not opportunistic pathogens and only a limited number cause food-borne illnesses, these results demonstrate that some B. cereus strains can cause severe and even fatal infections in patients who appear to be otherwise healthy.

C6786, the clinical isolate of the ‘Oklahoma’ strain of Pseudomonas (now Burkholderia) pseudomallei, was originally isolated in 1973 from a wound infection resulting from a farming accident in Oklahoma, USA. Environmental isolates C7532 and C7533 from the Oklahoma accident site were found to match C6786. These three isolates and a clinical isolate originally identified as B. pseudomallei that was recovered from a person in Georgia, USA, involved in an automobile accident were characterized by biochemical, 16S rRNA gene sequencing, multilocus sequence typing and DNA–DNA hybridization analyses. Results indicated that these strains comprise a novel species. The name Burkholderia oklahomensis sp. nov. is proposed, with strain C6786T (=LMG 23618T=NCTC 13387T=CCUG 51349T) as the type strain.

Cysticercosis has become increasingly recognized as an important infection in the United States in recent decades. Despite its potential impact, there is a lack of comprehensive information on the nationwide burden of disease. To better define the burden of cysticercosis in the United States, we analyzed in-patient records using the Nationwide Inpatient Sample for 1998-2011 to estimate cysticercosis-related hospitalizations and patient/institutional characteristics. There were an estimated 33,060 (95% confidence interval [95% CI] = 29,610.5-36,510.3) cysticercosis-related hospitalizations nationwide, representing a hospitalization rate of 8.03 per million population. The highest proportion of cases were male (54.8%), Hispanic (62.0%), aged 18-44 (58.8%), and occurred in the West (45.1%). An estimated 459 deaths occurred, representing an in-hospital case-fatality rate of 1.4%. These findings indicate the burden of cysticercosis-related hospitalizations in the United States is considerable and may be greater than currently appreciated. Cysticercosis should be a nationally reportable disease.

Adult and larval stages of Taenia solium cause 2 diseases in humans, i.e., taeniasis and cysticercosis, respectively. Diagnosis and treatment of taeniasis are the ultimate means to eliminate cysticercosis. A serological taeniasis diagnostic test has been developed for laboratory use. However, recombinant forms of the taeniasis diagnostic proteins are required to overcome the limited supply of native proteins and allow the development of a low-cost and field-applicable test with high sensitivity and specificity. Using 2-dimensional electrophoresis of T. solium excretory and secretory (TSES) products from hamster adult tape-worm in vitro cultures, we have identified 5 T. solium-specific protein spots, with molecular weights of 33 kDa (protein isoelectrofocusing point [pI]: 5.6, 5.3, 5.1) and 38 kDa (pI: 4.6, 4.5). Protein sequencing and molecular cloning of these proteins showed that although endowed with different pls, the proteins with the same molecular weights shared the same protein backbone, named TSES33 and TSES38. Their full-length complementary DNAs encode proteins with 267 and 278 amino acids, respectively. TSES33 and TSES38 were expressed in a baculovirus system. Both recombinant proteins were recognized by a panel of taeniasis, but not cysticercosis patient serum samples, indicating that they can potentially replace the native proteins in the development of a more efficacious taeniasis diagnostic test.

There is an increasing demand for diagnostic testing for Giardia intestinalis (G. lamblia) and Cryptosporidium parvum , with a priority being placed on obtaining diagnostic results in an efficient and timely manner. Several commercial companies have developed rapid diagnostic tests that are simple to perform and can be completed in less time than traditional methods for detecting Giardia and Cryptosporidium . We compared one of these rapid tests, the ImmunoCard STAT! (Meridian Bioscience, Inc.) lateral-flow immunoassay, with the MERIFLUOR direct fluorescent-antibody (DFA) test, the ProSpecT EZ microplate assay for Giardia and the ProSpecT microplate assay for Cryptosporidium , and modified Kinyoun's acid-fast stained smears for the detection of Cryptosporidium using 246 specimens. The MERIFLUOR DFA (Meridian Bioscience, Inc.) test detected the largest number of cases (32 Giardia and 37 Cryptosporidium ) infections and was used to calculate the sensitivity and specificity of the other tests. For Giardia , the sensitivities of the ImmunoCard STAT! and the ProSpecT Giardia EZ microplate assay (Alexon-Trend, Inc.) were 81 and 91%, respectively. For detection of Cryptosporidium , the sensitivities of the ImmunoCard STAT!, the ProSpecT Cryptosporidium microplate assay (Alexon-Trend, Inc.), and modified Kinyoun's acid-fast stained smears were 68, 70, and 78%, respectively. Test specificities were equal to or greater than 99%. Specimens with very small numbers of organisms were not detected by the ImmunoCard STAT!, the ProSpecT microplate assay or modified Kinyoun's acid-fast stained smears.

Background Strongyloidiasis is a persistent human parasitic infection caused by the intestinal nematode, Strongyloides stercoralis. The parasite has a world-wide distribution, particularly in tropical and subtropical regions with poor sanitary conditions. Since individuals with strongyloidiasis are typically asymptomatic, the infection can persist for decades without detection. Problems arise when individuals with unrecognized S. stercoralis infection are immunosuppressed, which can lead to hyper-infection syndrome and disseminated disease with an associated high mortality if untreated. Therefore a rapid, sensitive and easy to use method of diagnosing Strongyloides infection may improve the clinical management of this disease. Methodology/Principal Findings An immunological assay for diagnosing strongyloidiasis was developed on a novel diffraction-based optical bionsensor technology. The test employs a 31-kDa recombinant antigen called NIE derived from Strongyloides stercoralis L3-stage larvae. Assay performance was tested using retrospectively collected sera from patients with parasitologically confirmed strongyloidiasis and control sera from healthy individuals or those with other parasitoses including schistosomiasis, trichinosis, echinococcosis or amebiasis who were seronegative using the NIE ELISA assay. If we consider the control group as the true negative group, the assay readily differentiated S. stercoralis-infected patients from controls detecting 96.3% of the positive cases, and with no cross reactivity observed in the control group These results were in excellent agreement (κ = 0.98) with results obtained by an NIE-based enzyme-linked immunosorbent assay (ELISA). A further 44 sera from patients with suspected S. stercoralis infection were analyzed and showed 91% agreement with the NIE ELISA. Conclusions/Significance In summary, this test provides high sensitivity detection of serum IgG against the NIE Strongyloides antigen. The assay is easy to perform and provides results in less than 30 minutes, making this platform amenable to rapid near-patient screening with minimal technical expertise., Author Summary A rapid and sensitive serodiagnostic assay for strongyloidiasis based on a 31-kDa recombinant antigen from Strongyloides stercoralis (NIE) was developed using a novel diffraction-based optical biosensor technology. Assay performance was tested using retrospectively collected sera from patients with parasitologically confirmed strongyloidiasis (n = 54) and control sera from healthy individuals (n = 7) or those with other parasitoses including schistosomiasis, trichinosis, echinococcosis or amebiasis (n = 40). If we consider the control group as the true negative group, the assay readily differentiated S. stercoralis-infected patients from controls detecting 96.3% of the positive cases, and with no cross reactivity observed in the control group. These results were in excellent agreement (κ = 0.98) with results obtained by an NIE-based enzyme-linked immunosorbent assay (ELISA). A further 44 sera from patients with suspected S. stercoralis infection were analyzed and showed 91% agreement with the NIE ELISA. This test provides high sensitivity detection of serum IgG against the NIE Strongyloides antigen. The assay is easy to perform and provides results in less than 30 minutes, making this platform amenable to rapid near-patient screening with minimal technical expertise.

Toxoplasma gondii is a ubiquitous parasite that can cause neurologic and ocular disease. We tested sera from 7,072 people ≥ 6 years of age in the 2009–2010 National Health and Nutrition Examination Survey (NHANES) for immunoglobulin G antibodies and compared these results with two previous NHANES studies. The overall T. gondii antibody seroprevalence among persons ≥ 6 years of age in 2009–2010 was 13.2% (95% confidence limit [CL] 11.8%, 14.5%) and age-adjusted seroprevalence was 12.4% (95% CL 11.1%, 13.7%); age-adjusted seroprevalence among women 15–44 years of age was 9.1% (95% CL 7.2%, 11.1%). In U.S. born persons 12–49 years of age, the age-adjusted T. gondii seroprevalence decreased from 14.1% (95% CL 12.7%, 15.5%) in NHANES III (1988–1994) to 9.0% (95% CL 7.6%, 10.5%) in NHANES 1999–2004 to 6.7% (95% CL 5.3%, 8.2%) in NHANES 2009–2010 (P < 0.001 linear trend). Although T. gondii antibody presence is still relatively common, the prevalence in the United States has continued to decline.

Neglected parasitic infections, including Chagas disease, toxocariasis, cysticercosis, and toxoplasmosis, affect millions of persons in the United States. Relatively few resources have been devoted to surveillance, prevention, and treatment of these diseases. Chagas disease primarily affects Latin American immigrants and can cause heart failure and death if not treated. Immediate antiparasitic treatment is indicated for most patients with acute Chagas disease. Treatment is recommended for patients younger than 18 years who have chronic Chagas disease and is generally recommended for adults younger than 50 years who do not have advanced cardiomyopathy; treatment decisions for other patients should be made on an individual basis. Toxocariasis primarily affects children and can cause gastrointestinal, respiratory, and ophthalmologic disease. Treatment options include albendazole and mebendazole. Patients with ocular infection require referral to an ophthalmologist. Neurocysticercosis, a form of cysticercosis, is the most common infectious cause of seizures in some parts of the United States. Initial treatment should focus on symptom control. Humans generally acquire toxoplasmosis by eating undercooked contaminated meat or ingesting things that have been contaminated with cat feces. Congenital infection can result in miscarriage or adverse fetal effects. Treatment is recommended for immunosuppressed persons, pregnant women, and immunocompetent persons with severe symptoms.