Comparison Between Elisa and Dye Test for Detection of Naturally Acquired Toxoplasma Gondii Antibodies in the Goat

Toxoplasma gondii IgG antibodies were measured in 212 goat sera, comparing the Sabin-Feldman dye-test and a three-layer sandwich enzyme-linked immunosorbent assay (ELISA). With 98 % concordance obtained between these 2 tests, the results are at the same paragon as for human sera. Accordingly, the ELISA sandwich procedure appears to be suitable for large-scale analysis of goat sera. The discordant 2 % were ELISA positive and dye-test negative. One possible explanation of the divergent titres is given using an immunized goat model.     

Plate Hemolysin Test for the Rapid Screening of Toxoplasma Antibodies

A plate hemolysin test was developed to screen serum specimens for the presence of toxoplasma antibodies. When we tested 130 sera by both this test and the standard toxoplasma dye test, we found the plate hemolysin test to be a rapid, sensitive, and economical method for detecting toxoplasma antibodies. In all but one instance it paralleled the dye test. A comparison of the results of testing six sera by the hemolysin, hemagglutination, and dye-test techniques suggested that the hemolytic antibodies were more closely related to hemagglutinating antibodies than to dye-test antibodies. We could not store sheep erythrocytes sensitized with toxoplasma lysate for more than 3 days without altering the sensitivity of the test. Concanavalin A proved to be an effective coupling agent for binding toxoplasma antigens to red-cell membranes, a quality attributed to its affinity for specific polysaccharide-combining sites.     

An Epidemiologic Study of 100 Cases of Toxoplasmic Uveitis: Implications for Diagnosis and Prevention

An epidemiologic study of 100 patients with active chorioretinitis, positive Sabin-Feldman methylene-blue dye-test, and favourable response to antitoxoplasmic drugs (spiramycin) was undertaken to establish a basis for rapid clinical diagnosis and institution of preventive measures for toxoplasmic uveitis. Eighty-two of the 100 patients, but only 19 of 100 controls, gave positive histories of exposure to likely sources of toxoplasmosis, such as hunting and handling of wild game, ingestion of raw meat, etc. Seventy-six of these 82 patients had positive toxoplasmin skin tests, but all subsequently had positive dye-tests. The important practical conclusion was reached that all cases with active chorioretinitis, positive toxoplasmin skin test, or history of exposure to known reservoirs of toxoplasmosis should be treated immediately, as dye-tests in such cases will almost invariably be positive. This clinical approach greatly improves the visual prognosis of toxoplasmic chorioretinitis, since specific therapy need not be delayed for several weeks until the result of the dye-test is known.     

Potential risk factors associated with seropositivity for Toxoplasma gondii among pregnant women and HIV infected individuals in Ethiopia: A systematic review and meta-analysis

BackgroundToxoplasma gondii is an obligate intracellular and neurotropic apicomplexan protozoan parasite infecting almost all warm-blooded vertebrates including humans. To date in Ethiopia, no systematic study has been investigated on the overall effects of potential risk factors associated with seropositivity for Toxoplasma gondii among pregnant women and HIV infected individuals. We intended to determine the potential risk factors (PRFs) associated with seropositivity for Toxoplasma gondii from published data among pregnant women and HIV infected individuals of Ethiopia.MethodologyAn systematic review of the previous reports was made. We searched PubMed, Science Direct, African Journals Online, and Google Scholar for studies with no restriction on the year of publication. All references were screened independently in duplicate and were included if they presented data on at least two risk factors. Meta-analysis using the random or fixed-effects model was made to calculate the overall effects for each exposure.ResultsOf the 216 records identified, twenty-four reports met our eligibility criteria, with a total of 6003 individuals (4356 pregnant women and 1647 HIV infected individuals). The pooled prevalences of anti-Toxoplasma gondii antibodies were found at 72.5% (95% CI: 58.7% - 83.1%) in pregnant women and 85.7% (95% CI: 76.3% - 91.8%) in HIV infected individuals. A significant overall effect of anti-Toxoplasma gondii seropositivity among pregnant women (p < 0.05) was witnessed with age, abortion history, contact with cats, cat ownership, having knowledge about toxoplasmosis, being a housewife and having unsafe water source. Age, cat ownership, and raw meat consumption were also shown a significant effect (p < 0.05) to anti-Toxoplasma gondii seropositivity among HIV infected individuals.ConclusionsThis review showed gaps and drawbacks in the earlier studies that are useful to keep in mind to design accurate investigations in the future. The pooled prevalence of anti-Toxoplasma gondii antibodies was found to be higher among pregnant women and HIV infected individuals. This suggests that thousands of immunocompromised individuals (pregnant women and HIV infected patients) are at risk of toxoplasmosis due to the sociocultural and living standards of the communities of Ethiopia. Appropriate preventive measures are needed to reduce the exposure to Toxoplasma gondii infection. Further studies to investigate important risk factors are recommended to support the development of more cost-effective preventive strategies.     

Virulence without catalysis: how can a pseudokinase affect host cell signaling?

A hallmark of the pathogenic lifestyle is the secretion of enzymes and other effectors that dysregulate host signaling. Intriguingly, the most potent virulence locus identified in the intracellular parasite Toxoplasma gondii encodes a family of related catalytically inactive protein kinases, or pseudokinases. Toxoplasma has in its kinome among the highest percentage of pseudokinases among all sequenced organisms, and the majority of these appear to be secreted into the host cell. We posit that the pseudokinase fold represents a particularly well-suited domain for functional diversification, discuss the relevance of gene expansion at these loci, and outline potential mechanisms by which a pseudokinase might affect host signaling.     

Hematology of Southern Beaufort Sea Polar Bears (2005–2007): Biomarker for an Arctic Ecosystem Health Sentinel

Arctic temperatures are increasing in response to greenhouse gas forcing and polar bears have already responded to changing conditions. Declines in body stature and vital rates have been linked to warming-induced loss of sea-ice. As food webs change and human activities respond to a milder Arctic, exposure of polar bears and other arctic marine organisms to infectious agents may increase. Because of the polar bear’s status as arctic ecosystem sentinel, polar bear health could provide an index of changing pathogen occurrence throughout the Arctic, however, exposure and monitoring protocols have yet to be established. We examine prevalence of antibodies to Toxoplasma gondii, and four morbilliviruses (canine distemper [CDV], phocine distemper [PDV], dolphin morbillivirus [DMV], porpoise morbillivirus [PMV]) including risk factors for exposure. We also examine the relationships between antibody levels and hematologic values established in the previous companion article. Antibodies to Toxoplasma gondii and morbilliviruses were found in both sample years. We found a significant inverse relationship between CDV titer and total leukocytes, neutrophils, monocytes, and eosinophils, and a significant positive relationship between eosinophils and Toxoplasma gondii antibodies. Morbilliviral prevalence varied significantly among age cohorts, with 1–2 year olds least likely to be seropositive and bears aged 5–7 most likely. Data suggest that the presence of CDV and Toxoplasma gondii antibodies is associated with polar bear hematologic values. We conclude that exposure to CDV-like antigen is not randomly distributed among age classes and suggest that differing behaviors among life history stages may drive probability of specific antibody presence.     

Morbillivirus and <Emphasis Type="Italic">Toxoplasma</Emphasis> Exposure and Association with Hematological Parameters for Southern Beaufort Sea Polar Bears: Potential Response to Infectious Agents in a Sentinel Species

Arctic temperatures are increasing in response to greenhouse gas forcing and polar bears have already responded to changing conditions. Declines in body stature and vital rates have been linked to warming-induced loss of sea-ice. As food webs change and human activities respond to a milder Arctic, exposure of polar bears and other arctic marine organisms to infectious agents may increase. Because of the polar bear’s status as arctic ecosystem sentinel, polar bear health could provide an index of changing pathogen occurrence throughout the Arctic, however, exposure and monitoring protocols have yet to be established. We examine prevalence of antibodies to Toxoplasma gondii, and four morbilliviruses (canine distemper [CDV], phocine distemper [PDV], dolphin morbillivirus [DMV], porpoise morbillivirus [PMV]) including risk factors for exposure. We also examine the relationships between antibody levels and hematologic values established in the previous companion article. Antibodies to Toxoplasma gondii and morbilliviruses were found in both sample years. We found a significant inverse relationship between CDV titer and total leukocytes, neutrophils, monocytes, and eosinophils, and a significant positive relationship between eosinophils and Toxoplasma gondii antibodies. Morbilliviral prevalence varied significantly among age cohorts, with 1–2 year olds least likely to be seropositive and bears aged 5–7 most likely. Data suggest that the presence of CDV and Toxoplasma gondii antibodies is associated with polar bear hematologic values. We conclude that exposure to CDV-like antigen is not randomly distributed among age classes and suggest that differing behaviors among life history stages may drive probability of specific antibody presence.     

Structure of the immunodominant surface antigen from the Toxoplasma gondii SRS superfamily

Toxoplasma gondii is a persistent protozoan parasite capable of infecting almost any warm-blooded vertebrate. The surface of Toxoplasma is coated with a family of developmentally regulated glycosylphosphatidylinositol (GPI)-linked proteins (SRSs), of which SAG1 is the prototypic member. SRS proteins mediate attachment to host cells and interface with the host immune response to regulate the virulence of the parasite. The 1.7 A structure of the immunodominant SAG1 antigen reveals a homodimeric configuration in which the dimeric interface is mediated by an extended beta-sheet that forms a deep groove lined with positively charged amino acids. This basic groove seems to be conserved among SRS proteins and potentially serves as a sulfated proteoglycan-binding site on target cell surfaces, thus rationalizing the promiscuous attachment properties of Toxoplasma to a broad range of host cell types.     

Toxoplasma gondii triggers secretion of interleukin-12 but low level of interleukin-10 from the THP-1 human monocytic cell line

Previous studies using both in vitro and in vivo mouse models have demonstrated that a subtle balance between pro- and anti-inflammatory cytokines, among which interleukin-12 (IL-12) and interleukin-10 (IL-10), respectively is crucial to control Toxoplasma infection. However, the few studies performed with human cell lines highlighted important host-related differences in the immune response to Toxoplasma gondii. The goal of our work was thus to study the production of both IL-12 and IL-10 by the THP-1 human monocytic cell line in response to Toxoplasma. We demonstrated that infection by live parasites (RH strain) triggers secretion of IL-12, but low level of IL-10. IL-12 secretion appeared within 8 h, up to 48 h. We also showed that infection by live parasites is not mandatory since heat-killed parasites, crude tachyzoite lysate as well as excreted/secreted antigens induced significant, yet reduced production of IL-12.     

Sex-dependent toxoplasmosis-associated differences in testosterone concentration in humans

Several lines of indirect evidence suggest that subjects with latent infection of the coccidian parasite Toxoplasma gondii have a higher concentration of testosterone than uninfected controls. Here, we searched for direct evidence of latent toxoplasmosis-associated differences in testosterone concentration among a population of 174 female and 91 male students screened for Toxoplasma infection. We have found Toxoplasma-infected men to have a higher concentration of testosterone and Toxoplasma-infected women to have a lower concentration of testosterone than Toxoplasma-free controls. The opposite direction of the testosterone shift in men compared to women can explain the observed gender specificity of behavioural shifts in Toxoplasma-infected subjects.     

Unique differences in infectivity and seroreactivity of Toxoplasma harvested from mice infected for different lengths of time

For use in experiments, Toxoplasma of the RH strain are usually harvested from mouse peritoneal cavities 48 hr (2-day Toxoplasma) or more after intraperitoneal inoculation. In this report we show that Toxoplasma harvested at 24 hr (1-day Toxoplasma) after inoculation are much more infective for and replicate to a greater degree within mouse resident peritoneal macrophages in vitro and are much more resistant to the cidal activity of activated mouse peritoneal macrophages and resident rat peritoneal macrophages than are 2-day Toxoplasma. Ingestion of 1-day Toxoplasma by macrophages did not trigger the respiratory burst as measured by reduction of nitroblue tetrazolium (NBT), but coating 1-day Toxoplasma with specific antibody did result in reduced NBT. However, coating 1-day Toxoplasma with specific antibody did not markedly decrease infectivity for macrophages in vitro, unlike decreased infectivity observed when 2-day Toxoplasma are coated with specific antibody. Use of 1-day Toxoplasma in the dye test resulted in a 5-fold decrease in titer of specific antibody in human sera. Use of Toxoplasma harvested 24 hr after infection may serve as a new tool to probe virulence factors of Toxoplasma and of host cells' antimicrobial mechanisms.     

Importance of endogenous IFN-gamma for prevention of toxoplasmic encephalitis in mice

The importance of endogenous IFN-gamma for prevention of toxoplasmic encephalitis was studied in mice chronically infected with Toxoplasma gondii by using a mAb to this lymphokine. Control mice chronically infected with the ME49 strain that received saline or normal IgG had slight inflammation in their brains whereas those that received the mAb developed severe encephalitis. In contrast to control mice, the mAb-treated mice had many areas of acute focal inflammation and infiltration of large numbers of inflammatory cells in the meninges and parenchyma of their brains. In the areas of acute focal inflammation, tachyzoites and Toxoplasma Ag were demonstrated by immunoperoxidase staining with the use of rabbit anti-Toxoplasma antibody, suggesting that the focal inflammation was induced by Toxoplasma organisms. Acute inflammation was also observed around cysts of Toxoplasma. Immunohistologic staining revealed tachyzoites and Toxoplasma Ag surrounding the periphery of these cysts suggesting cyst disruption had occurred. Mice treated with mAb against IFN-gamma had five times the numbers of cysts in their brains as did control mice. These results clearly indicate that endogenous IFN-gamma plays a significant and important role in prevention of encephalitis in mice chronically infected with Toxoplasma. The mAb-treated mice had the same Toxoplasma antibody titers and the same degree of macrophage killing of Toxoplasma as did untreated controls. These results suggest that IFN-gamma may have a direct role in preventing cyst rupture and toxoplasmic encephalitis.     

Critical analysis of the topology and rooting of the parabasalian 16S rRNA tree

Between 20% and 60% of the population of most countries are infected with the protozoan Toxoplasma gondii . Subjects with clinically asymptomatic life-long latent toxoplasmosis differ from those who are Toxoplasma free in several behavioral parameters. Case–control studies cannot decide whether these differences already existed before infection or whether they were induced by the presence of Toxoplasma in the brain of infected hosts. Here, we searched for such morphological differences between Toxoxoplasma -infected and Toxoplasma -free subjects that could be induced by the parasite (body weight, body height, body mass index, waist–hip ratio), or could rather correlate with their natural resistance to parasitic infection (fluctuating asymmetry, 2D : 4D ratio). We found Toxoplasma -infected men to be taller and Toxoplasma -infected men and women to have lower 2D : 4D ratios previously reported to be associated with higher prenatal testosterone levels. The 2D : 4D ratio negatively correlated with the level of specific anti- Toxoplasma antibodies in Toxoplasma -free subjects. These results suggest that some of the observed differences between infected and noninfected subjects may have existed before infection and could be caused by the lower natural resistance to Toxoplasma infection in subjects with higher prenatal testosterone levels.     

Toxoplasma polymorphic effectors determine macrophage polarization and intestinal inflammation

European and North American strains of the parasite Toxoplasma gondii belong to three distinct clonal lineages, type I, type II, and type III, which differ in virulence. Understanding the basis of Toxoplasma strain differences and how secreted effectors work to achieve chronic infection is a major goal of current research. Here we show that type I and III infected macrophages, a cell type required for host immunity to Toxoplasma, are alternatively activated, while type II infected macrophages are classically activated. The Toxoplasma rhoptry kinase ROP16, which activates STAT6, is responsible for alternative activation. The Toxoplasma dense granule protein GRA15, which activates NF-κB, promotes classical activation by type II parasites. These effectors antagonistically regulate many of the same genes, and mice infected with type II parasites expressing type I ROP16 are?protected against Toxoplasma-induced ileitis. Thus, polymorphisms in determinants that modulate?macrophage activation influence the ability of Toxoplasma to establish a chronic infection.     

Toxoplasma modifies macrophage phagosomes by secretion of a vesicular network rich in surface proteins

Modification of macrophage phagosomes begins shortly after formation as Toxoplasma cells secrete membranous vesicles that form a reticulate network within the vacuole. The Toxoplasma-modified compartments then resist normal endocytic processing and digestion. We have used the pronounced Ca++-dependent stability of the intraphagosomal membrane (IPM) network to purify and characterize the structural proteins of this assembly. In addition to the structural matrix, Toxoplasma secretes a discrete set of soluble proteins, including a newly described 22-kD calcium-binding protein. The IPM network adheres to intact Toxoplasma cells after host cell lysis in the presence of 1 mM Ca++; however, the network readily disperses in calcium-free buffer and was purified as vesicles that sedimented at 100,000 g. Purified IPM vesicles were specifically recognized by immune sera from mice with chronic Toxoplasma infection and consisted primarily of a 30-kD protein when analyzed by SDS PAGE. IPM network proteins share a major antigenic component located on the surface of extracellular Toxoplasma cells as shown by immunoperoxidase electron microscopy using a polyclonal antibody prepared against the IPM vesicles. Moreover, in Toxoplasma-infected macrophages, anti-IMP antibody confirmed that the extensive IPM array contains proteins also found on the Toxoplasma cell surface. Our results indicate the IMP network represents a unique structural modification of the phagosome comprised in part of Toxoplasma surface proteins.     

Predator cat odors activate sexual arousal pathways in brains of Toxoplasma gondii infected rats

Cat odors induce rapid, innate and stereotyped defensive behaviors in rats at first exposure, a presumed response to the evolutionary pressures of predation. Bizarrely, rats infected with the brain parasite Toxoplasma gondii approach the cat odors they typically avoid. Since the protozoan Toxoplasma requires the cat to sexually reproduce, this change in host behavior is thought to be a remarkable example of a parasite manipulating a mammalian host for its own benefit. Toxoplasma does not influence host response to non-feline predator odor nor does it alter behavior on olfactory, social, fear or anxiety tests, arguing for specific manipulation in the processing of cat odor. We report that Toxoplasma infection alters neural activity in limbic brain areas necessary for innate defensive behavior in response to cat odor. Moreover, Toxoplasma increases activity in nearby limbic regions of sexual attraction when the rat is exposed to cat urine, compelling evidence that Toxoplasma overwhelms the innate fear response by causing, in its stead, a type of sexual attraction to the normally aversive cat odor.     

Tazonomy of Toxoplasma.

After reviewing reports of the hosts, structure and life cycle of Toxoplasma, the genus is placed in the apicomplexan family Eimeriidae and the folllowing 7 species are recognized: Toxoplasma gondii (Nicolle & Manceaux) (type species) from about 200 species of mammals and birds, with oocysts in felids; Toxoplasma alencari (Da Costa & Pereira) from the frog Leptodactylus ocellatus; Toxoplasma brumpti Coutelen from the iguana Iguana tuberculata; Toxoplasma colubri Tibaldi from the snakes Coluber melanoleucus and Coluber viridiflavus; Toxoplasma hammondi (Frenkel & Dubey) (a new combination for Hammondia hammondi) from the house mouse with oocysts in the domestic cat; Toxoplasma ranae Levine & Nye from the leopard frog Rana Pipiens; and Toxoplasma serpai Scorza, Dagert & Iturriza Arocha from the toad Bufo marinus.     

实验动物血液及精液感染弓形虫动态研究

采用荧光定量PCR技术对实验动物家兔血液及精液感染弓形虫DNA检测实验及动态变化研究,阐明了弓形虫感染的发展和转归机理,为了解弓形虫与宿主动态关系,弓形虫经血液、精液传播提供理论依据.对性传播疾病、特别是艾滋病的预防提供重要参考价值.