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1.  Toxoplasma gondii infection in interstate truck drivers: a case–control seroprevalence study 
Parasites & Vectors  2015;8:77.
Background
Infection with Toxoplasma gondii can be acquired via the ingestion of undercooked or raw meat containing tissue cysts, or via ingestion of water contaminated with oocysts. Professional long distance truck driving may have epidemiological importance for food-borne infections since drivers eat out of home and in places where hygiene and cooking practices are uncertain. We aimed to determine whether interstate truck drivers in Durango, Mexico have an increased risk of infection with T. gondii as indicated by seropositivity; and to determine the socio-demographic, work, clinical, and behavioral characteristics associated with T. gondii seropositivity in interstate truck drivers.
Methods
Through a case–control study design, 192 truck drivers and 192 controls from the general population of the same region matched by gender and age were examined with enzyme-linked immunoassays for the presence of anti-Toxoplasma IgG and IgM antibodies. Socio-demographic, work, clinical and behavioral characteristics from the truck drivers were obtained.
Results
Anti-T. gondii IgG antibodies were found in 23 (12.0%) of 192 truck drivers and in 13 (6.8%) of 192 controls (OR = 21.0; 95% CI: 1.23-358.38; P = 0.002). Anti-T. gondii IgM antibodies were found in 7 (3.6%) cases and in 7 (3.6%) controls (P = 1.00). The seroprevalence of T. gondii infection was higher in drivers with reflex impairment than in those without this impairment (4/13, 30.8% vs 19/179, 10.6%, respectively; P = 0.05), and in drivers with hearing impairment than in those without this impairment (3/7, 42.9% vs 20/185, 10.8%, respectively; P = 0.03). Multivariate analysis of work and behavioral characteristics of truck drives showed positive associations of T. gondii exposure with trips to the south of Mexico (OR = 3.11; 95% CI: 1.02-9.44; P = 0.04) and consumption of horse meat (OR = 5.18; 95% CI: 1.62-16.55; P = 0.005).
Conclusions
Results suggest that interstate truck drivers may have an increased risk for T. gondii infection, and that T. gondii exposure may be impacting neurological functions in truck drivers. Contributing factors for T. gondii exposure should be taken into account for the design of optimal prevention measures against T. gondii infection.
doi:10.1186/s13071-015-0690-z
PMCID: PMC4322546  PMID: 25651797
Toxoplasma gondii; Seroprevalence; Truck drivers; Case–control study; Risk factors
2.  Lack of association between Toxoplasma gondii infection and occupational exposure to animals 
The association of infection with Toxoplasma gondii and occupational exposure to animals has been scantly determined. We performed a case-control study with 200 subjects from Durango Province, Mexico, occupationally exposed to animals and 200 age- and gender-matched subjects without this occupation. Sera from all participants were analyzed for anti-T. gondii IgG and IgM antibodies using enzyme-linked immunoassays. The association of seroprevalence with sociodemographic, work, clinical, and behavioral characteristics in cases was determined.
Cases and controls had similar frequencies of anti-T. gondii IgG antibodies (12/200: 6.0% and 11/200: 5.5%, respectively) (OR = 3.0; 95% CI: 0.12–73.64; P = 1.0). The frequency of sera with high (>150 IU/ml) levels of anti-T. gondii IgG antibodies was comparable among cases and controls (P = 0.61). Seroprevalence of anti-T. gondii IgM antibodies was similar in cases (4, 2.0%) than in controls (4, 2.0%) (P = 1.0). Multivariate analysis showed that seropositivity was associated with eating while working (OR = 7.14; 95% CI: 1.91–26.72; P = 0.003) and consumption of duck meat (OR = 5.43; 95% CI: 1.43–20.54; P = 0.01).
No association between seropositivity to T. gondii and occupational exposure to animals was found. However, risk factors for infection found should be taken into account to reduce the exposure to T. gondii.
doi:10.1556/EUJMI-D-14-00024
PMCID: PMC4271814  PMID: 25544890
animals; epidemiology; infection; Mexico; occupational exposure; seroprevalence; Toxoplasma gondii
3.  Toxoplasma Serotype Is Associated With Development of Ocular Toxoplasmosis 
The Journal of Infectious Diseases  2013;208(9):1520-1528.
Background. Worldwide, ocular toxoplasmosis (OT) is the principal cause of posterior uveitis, a severe, life-altering disease. A Toxoplasma gondii enzyme-linked immunoassay that detects strain-specific antibodies present in serum was used to correlate serotype with disease.
Methods. Toxoplasma serotypes in consecutive serum samples from German uveitis patients with OT were compared with non-OT seropositive patients with noninfectious autoimmune posterior uveitis. OT patients were tested for association of parasite serotype with age, gender, location, clinical onset, size, visual acuity, or number of lesions (mean follow-up, 3.8 years) to determine association with recurrences.
Results. A novel, nonreactive (NR) serotype was detected more frequently in serum samples of OT patients (50/114, 44%) than in non-OT patients (4/56, 7%) (odds ratio, 10.0; 95% confidence interval 3.4–40.8; P < .0001). Non-OT patients were predominantly infected with Type II strains (39/56; 70%), consistent with expected frequencies in Central Europe. Among OT patients, those with NR serotypes experienced more frequent recurrences (P = .037). Polymerase chain reaction detected parasite DNA in 8/60 OT aqueous humor specimens but failed to identify Type II strain alleles.
Conclusions. Toxoplasma NR and Type II serotypes predominate in German OT patients. The NR serotype is associated with OT recurrences, underscoring the value of screening for management of disease.
doi:10.1093/infdis/jit313
PMCID: PMC3789564  PMID: 23878321
Diagnosis; ocular inflammatory disease; serotype; toxoplasmosis; uveitis
4.  Intracellular transport of Toxoplasma gondii through the blood–brain barrier 
Journal of neuroimmunology  2010;232(0):119-130.
Toxoplasma gondii establishes latent infection in the central nervous system of immunocompentent hosts. Toxoplasmic encephalitis is a life threatening reactivation of latent infection in the brain of immunocompromised patients. To further understand the mechanisms of entry into the brain of T. gondii we investigated host molecules and cells involved in the passage of the parasite through the blood–brain barrier. First, using microarrays brain endothelial cells were found to upregulate, among others, chemokines and adhesion molecules following infection with tachyzoites. Using flow cytometry we observed upregulated ICAM-1 expression on the surface of brain endothelial cells following infection; ICAM-1 expression was further increased after pre-incubation with IFN-γ. Compared to RH tachyzoites, ME49 tachyzoites induced a stronger upregulation of ICAM-1 and an earlier and stronger IL-6 and MCP-1 secretion by brain endothelial cells. Using an in vitro coculture model of the BBB (primary glia cells and brain endothelial cells) we found a stronger migration of infected antigen-presenting cells compared to lymphocytes (4.63% vs. 0.6% of all cells) across the BBB. Among all antigen-presenting cells CD11b+/CD11c+ cells showed the highest infection rate, whereas the majority of infected cells that migrated through the blood–brain barrier were CD11b+/CD11c− cells. Infection of PBMCs with type I or type II Toxoplasma strains resulted in similar patterns of cell migration across the in vitro BBB model.
In conclusion, these results suggest that T. gondii modulates gene expression of brain endothelial cells to promote its own migration through the blood–brain barrier in a ‘Trojan horse’ manner. Cells expressing CD11b either with or without CD11c are likely candidate cells for the intracellular transport of T. gondii across the BBB. T. gondii type I and type II strains induced similar migration patterns of antigen-presenting cells across the in vitro BBB.
doi:10.1016/j.jneuroim.2010.10.029
PMCID: PMC4116595  PMID: 21106256
Toxoplasma gondii; Blood–brain barrier; Neuroinvasion; Transendothelial migration
5.  Seroprevalence of Toxoplasma gondii infection and associated risk factors in Huicholes in Mexico 
Parasites & Vectors  2014;7:301.
Background
Very little is known about the seroepidemiology of Toxoplasma gondii infection in ethnic groups in Mexico. Huicholes are an indigenous ethnic group living in a remote mountainous region in Mexico. We sought to determine the prevalence of anti-Toxoplasma IgG and IgM antibodies in Huicholes; and to determine the association of Toxoplasma seropositivity with socio-demographic, behavioral, and clinical characteristics of Huicholes.
Methods
We performed a cross sectional survey in Huicholes from September 2013 to January 2014. A convenience sampling method was used. We investigated the prevalence of anti-Toxoplasma IgG and IgM antibodies in 214 Huicholes using enzyme-linked immunoassays. A standardized questionnaire was used to obtain the characteristics of the Huicholes. Bivariate and multivariate analyses were used to assess the association of Toxoplasma exposure and Huicholes’ characteristics.
Results
Of the 214 Huicholes studied (mean age: 37.98 ± 15.80 years), 71 (33.2%) were positive for anti-T. gondii IgG antibodies and 47 (66.2%) of them were also positive for anti-T. gondii IgM antibodies. Seroprevalence of T. gondii infection did not vary with age, sex, or occupation. However, seroprevalence of anti-T. gondii IgM antibodies was significantly higher in female than in male Huicholes. Multivariate analysis of socio-demographic and behavioral characteristics showed that T. gondii exposure was associated with consumption of turkey meat (OR = 2.28; 95% CI: 1.16-4.46; P = 0.01). In addition, seroprevalence of T. gondii infection was significantly higher in Huicholes suffering from dizziness and memory impairment than those without such clinical characteristics.
Conclusions
Our results demonstrate serological evidence of T. gondii exposure among Huicholes which may be impacting their health. Results of this first study of T. gondii infection in Huicholes may be useful for the design of optimal preventive measures against infection with T. gondii.
doi:10.1186/1756-3305-7-301
PMCID: PMC4226977  PMID: 24984845
Toxoplasma gondii; Seroprevalence; Huicholes; Cross-sectional study
6.  The impact of Toll-like-receptor-9 on intestinal microbiota composition and extra-intestinal sequelae in experimental Toxoplasma gondii induced ileitis 
Gut Pathogens  2014;6:19.
Background
Following peroral Toxoplasma (T.) gondii infection, susceptible mice develop acute ileitis due to a microbiota-dependent Th1 type immunopathology. Toll-like-receptor (TLR)-9 is known to recognize bacterial DNA and mediates intestinal inflammation, but its impact on intestinal microbiota composition and extra-intestinal sequelae following T. gondii infection has not yet been elucidated.
Methods and results
Seven days following peroral infection (p.i.) with 100 cysts of T. gondii ME49 strain, TLR-9-/- and wildtype (WT) mice suffered from comparable ileitis, whereas ileal parasitic loads as well as IFN-γ and nitric oxide levels were higher in TLR-9-/- compared to WT mice. Locally, TLR-9-/- mice exhibited increased ileal CD3+, but not FOXP3+ cell numbers at day 7 p.i.; in mesenteric lymph nodes IFN-γ-producing CD4+ cell numbers and TNF-α and IFN-γ concentrations were also increased in TLR-9-/- compared to WT mice. T. gondii DNA levels, however, did not differ in mice of either genotype. Differences in intestinal microbiota were rather subtle except for bifidobacteria that were virtually absent in both, naïve and T. gondii infected TLR-9-/-, but not WT mice. Extra-intestinally, TLR-9-/- mice displayed less distinct systemic immune responses as indicated by lower serum IL-6, and splenic TNF-α and IFN-γ levels as compared to WT mice despite higher translocation rates of intestinal bacteria to extra-intestinal compartments such as liver, spleen, kidney, and cardiac blood. Most importantly, brains were also affected in this inflammatory scenario as early as day 7 p.i. Remarkably, TLR-9-/- mice exhibited more pronounced inflammatory infiltrates with higher numbers of F4/80+ macrophages and microglia in the cortex and meninges as compared to WT mice, whereas T. gondii DNA levels did not differ.
Conclusion
We here show that TLR-9 is not required for the development of T. gondii induced ileitis but mediates distinct inflammatory changes in intestinal and extra-intestinal compartments including the brain.
doi:10.1186/1757-4749-6-19
PMCID: PMC4057803  PMID: 24932221
Toxoplasma gondii; Acute ileitis; TLR-9; Th1-type immunopathology; Extra-intestinal immune responses; Intracerebral inflammation; Pro-inflammatory cytokines; Intestinal microbiota composition; Bacterial translocation; Systemic inflammatory response; Bifidobacteria; FOXP3; Regulatory T cells; Gut-brain-axis
7.  Lack of association between Toxoplasma gondii infection and hypertensive disorders in pregnancy: a case–control study in a Northern Mexican population 
Parasites & Vectors  2014;7:167.
Background
The outcome of pregnancy is often threatened by hypertension disorders, i.e. eclampsia. Rate of infection with the protozoan parasite Toxoplasma gondii can be as high as 80% in pregnant women, and infection acquired during pregnancy can lead to fetal death. Very little is known about a potential association between infections, i.e. those with Toxoplasma gondii, and hypertensive disorders during pregnancy.
Methods
Through a case–control study design, we investigated the presence of anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies in 146 pregnant women suffering from hypertensive disorders (cases) and 146 age-matched normotensive pregnant women (controls) attending a public hospital in Durango City, Mexico. Obstetric and blood pressure characteristics from cases and controls were also obtained.
Results
Seroprevalence of anti-Toxoplasma IgG antibodies and IgG titers did not differ significantly in controls (8/146; 5.5%) and cases (9/146; 6.2%). Anti-Toxoplasma IgM antibodies were found in 2 (1.2%) controls and none of the cases. Seroprevalence of T. gondii in controls (5.5%) was similar to seroprevalences found in patients with mild preeclampsia (4/27: 14.8%), severe preeclampsia (5/95: 5.3%), eclampsia (0/16: 0%) and HELLP syndrome (hemolysis, elevated liver enzymes, and low platelet count) (0/8: 0%) (P = 0.23).
Conclusions
Our results suggest that latent infection with T. gondii is not associated with hypertensive disorders in pregnant women in Northern Mexico. Further studies with larger sample sizes are needed to elucidate the association of infection with T. gondii with hypertensive disorders in pregnancy.
doi:10.1186/1756-3305-7-167
PMCID: PMC3976501  PMID: 24708729
Toxoplasma gondii; Seroprevalence; Preeclampsia; HELLP syndrome; Eclampsia; Infection; Epidemiology
8.  Regulation and Migratory Role of P-Selectin Ligands during Intestinal Inflammation 
PLoS ONE  2013;8(4):e62055.
Dendritic cells from mesenteric lymph nodes (MLN) can convert retinal to retinoic acid (RA), which promotes induction of the gut-specific homing receptor α4β7. In contrast, priming within peripheral lymph nodes leads to upregulation of E- and P-selectin ligands (E- and P-lig). Apart from its α4β7 promoting effect, RA was shown to suppress E- and P-lig induction in vitro. However, enhanced frequencies of P-lig+ CD4+ T cells were reported during intestinal inflammation. To understand this contradiction, we first determined whether location of intestinal inflammation, that is, ileitis or colitis, affects P-lig induction. Both conditions promoted P-lig expression on CD4+ T cells; however, P-lig expressed on T cells facilitated Th1 cell recruitment only into the inflamed colon but not into inflamed small intestine induced by oral Toxoplasma gondii infection. A majority of P-lig+CD4+ T cells found within MLN during intestinal inflammation co-expressed α4β7 confirming their activation in the presence of RA. Mesenteric P-lig+CD4+ cells co-expressed the 130 kDa isoform of CD43 which requires activity of core 2 (beta)1,6-N-acetyl-glycosaminyltransferase-I (C2GlcNAcT-I) suggesting that C2GlcNAcT-I contributes to P-lig expression under these conditions. To test whether inflammatory mediators can indeed overrule the inhibitory effect of RA on P-lig expression we stimulated CD4+ T cells either polyclonal in the presence of IL-12 and IFNγ or by LPS-activated MLN-derived dendritic cells. Both conditions promoted P-lig induction even in the presence of RA. While RA impeded the induction of fucosyltransferase-VII it did not affect IL-12-dependent C2GlcNAcT-I induction suggesting that C2GlcNAcT-I can support P-lig expression even if fucosyltransferase-VII mRNA upregulation is dampened.
doi:10.1371/journal.pone.0062055
PMCID: PMC3632518  PMID: 23630623
9.  Performance of the cobas CT/NG Test Compared to the Aptima AC2 and Viper CTQ/GCQ Assays for Detection of Chlamydia trachomatis and Neisseria gonorrhoeae 
Journal of Clinical Microbiology  2012;50(7):2244-2249.
The next-generation amplification test for Chlamydia trachomatis and Neisseria gonorrhoeae (Roche cobas 4800), a fully automated system, was compared head to head, using female samples, to Gen-Probe Aptima Combo 2 and BD ProbeTec using Viper. Endocervical swabs, female urine, and endocervical samples in liquid-based cytology medium were run on at least two of three platforms. A total of 4,316 samples were evaluated, and 281 chlamydial and 69 gonococcal infections were identified. Estimates of sensitivity and specificity were obtained relative to the patient infection standard (PIS) and using latent class analysis (LCA). Chlamydia sensitivity estimates ranged from 86.9 to 95.6% using PIS and 97.6 to 98% using LCA. Specificity was ≥99.6% for all sample types. Sensitivity ranged from 95.6 to 100% using PIS and 96.9 to 100% using LCA for the detection of gonococcal infections. Specificity for gonococcal infections was ≥99.8%. cobas 4800 performance was equivalent to the comparator assays (all P values, >0.05), and the fully automated system provides high laboratory efficiency.
doi:10.1128/JCM.06481-11
PMCID: PMC3405604  PMID: 22518864
10.  Toxoplasma gondii and the blood-brain barrier 
Virulence  2012;3(2):182-192.
Infection with the protozoan parasite Toxoplasma gondii is characterized by asymptomatic latent infection in the central nervous system and skeletal muscle tissue in the majority of immunocompentent individuals. Life-threatening reactivation of the infection in immunocompromized patients originates from rupture of Toxoplasma cysts in the brain. While major progress has been made in our understanding of the immunopathogenesis of infection the mechanism(s) of neuroinvasion of the parasite remains poorly understood. The present review presents the current understanding of blood-brain barrier (patho)physiology and the interaction of Toxoplasma gondii with cells of the blood-brain barrier.
doi:10.4161/viru.19004
PMCID: PMC3396697  PMID: 22460645
Toxoplasma gondii; astrocytes; blood-brain barrier; cytokines; endothelial cells
11.  High seroprevalence of Toxoplasma gondii infection in a subset of Mexican patients with work accidents and low socioeconomic status 
Parasites & Vectors  2012;5:13.
Background
Toxoplasma gondii has been associated with reflex impairment and traffic accidents. It is unknown whether Toxoplasma infection might be associated with work accidents. Therefore, using a case-control seroprevalence study design, 133 patients with a recent work accident and 266 control subjects of the general population from the same region were examined with enzyme-linked immunoassays for the presence and levels of anti-Toxoplasma IgG antibodies and anti-Toxoplasma IgM antibodies. Socio-demographic, work, clinical and behavioral characteristics from each worker were obtained.
Results
Eleven (8.3%) of 133 patients, and 14 (5.3%) of 266 controls had anti-T. gondii IgG antibodies. Anti-T. gondii IgG levels were higher than 150 IU/ml in 8 (6%) patients and 10 (3.8%) controls. Anti-T. gondii IgM antibodies were found in one (0.8%) of the workers, and in 6 (2.3%) of the controls. No statistically significant differences in the IgG seroprevalences, frequencies of high IgG levels, and IgM seroprevalences among patients and controls were found. In contrast, a low socio-economic level in patients with work accidents was associated with Toxoplasma seropositivity (P = 0.01). Patients with work accidents and low socioeconomic status showed a significantly (OR = 3.38; 95% CI: 0.84-16.06; P = 0.04) higher seroprevalence of T. gondii infection than controls of the same socioeconomic status (15.1% vs. 5%, respectively). Multivariate analysis showed a positive association of T. gondii infection with boar meat consumption (OR = 3.04; 95% CI: 1.03-8.94; P = 0.04). In contrast, a negative association between T. gondii infection and national trips (OR = 0.40; 95% CI: 0.17-0.96; P = 0.04), sausage consumption (OR = 0.20; 95% CI: 0.05-0.68; P = 0.01), and ham consumption (OR = 0.16; 95% CI: 0.05-0.51; P = 0.002) was found.
Conclusions
In the study described here seropositivity to T. gondii was associated to work accidents in a subset of patients with low socioeconomic status. This is the first report of an association of T. gondii infection and work accidents. Further studies to confirm our results are needed. Results may help in designing optimal prevention strategies to avoid T. gondii infection.
doi:10.1186/1756-3305-5-13
PMCID: PMC3287140  PMID: 22236539
Toxoplasma gondii; seroprevalence; work accidents; case-control study; epidemiology
12.  Toxoplasma gondii infection in workers occupationally exposed to unwashed raw fruits and vegetables: a case control seroprevalence study 
Parasites & Vectors  2011;4:235.
Background
Through a case control seroprevalence study, we sought to determine the association of Toxoplasma gondii infection with occupational exposure to unwashed raw fruits and vegetables.
Methods
Subjects, numbering 200, who worked growing or selling fruits and vegetables, and 400 control subjects matched by age, gender, and residence were examined by enzyme immunoassays for the presence of anti-Toxoplasma IgG and IgM antibodies. Socio-demographic, clinical, and behavioral characteristics from the study subjects were obtained.
Results
Of the 200 fruit and vegetable workers, 15 (7.5%) of whom, and 31 (7.8%) of the 400 controls were positive for anti-Toxoplasma IgG antibodies (P = 0.96). Anti-Toxoplasma IgM antibodies were found in 2 (1%) of the fruit workers and in 11 (2.8%) of the control subjects (P = 0.23). Seroprevalence of Toxoplasma antibodies increased with age (P = 0.0004). In addition, seropositivity to Toxoplasma was associated with ill status (P = 0.04), chronic tonsillitis (P = 0.03), and reflex impairment (P = 0.03). Multivariate analysis showed that Toxoplasma infection was associated with consumption of raw meat (OR = 5.77; 95% CI: 1.15-28.79; P = 0.03), unwashed raw fruits (OR = 2.50; 95% CI: 1.11-5.63; P = 0.02), and living in a house with soil floors (OR = 3.10; 95% CI: 1.22-7.88; P = 0.01), whereas Toxoplasma infection was negatively associated with traveling abroad (OR = 0.28; 95% CI: 0.12-0.67; P = 0.005).
Conclusions
This is the first report of seroprevalence and contributing factors for Toxoplasma infection in workers occupationally exposed to unwashed raw fruits and vegetables, and the results may help in the design of optimal preventive measures against Toxoplasma infection especially in female workers at reproductive age.
doi:10.1186/1756-3305-4-235
PMCID: PMC3259050  PMID: 22177118
Case-control study; fruits workers; occupational exposure; seroprevalence; epidemiology
13.  Prevalence and Risk Factors of Toxoplasmosis among Pregnant Women in Fortaleza, Northeastern Brazil 
We determined the seroprevalence of Toxoplasma gondii and associated risk factors among 963 pregnant women attending an obstetric hospital in Fortaleza, Brazil. Seroprevalences of IgG and IgM against T. gondii were 68.6% (95% confidence interval [CI] = 65.6–71.6%) and 0.5% (95% CI = 0.06–1.0%), respectively. Seroprevalence of IgG was high in women less than 25 years of age (91.7%) and in low-income women (odds Ratio [OR] = 1.40, 95% CI = 1.02–1.90). Multivariate regression analysis showed that consumption of homemade water ice (adjusted OR = 1.49, 95% CI = 1.09–2.04), vegetables washed with untreated water (adjusted OR = 1.43, 95% CI = 1.05–1.94), consumption of chicken (adjusted OR = 1.49, 95% CI = 1.12–2.0), and dog ownership (adjusted OR= 1.46, 95% CI = 1.07–1.98) were factors associated with IgG seropositivity. Young women in northeastern Brazil living under poor socioeconomic conditions are at highest risk for acquiring infection with T. gondii. Oocyst contamination of water and soil must be addressed in future prevention strategies.
doi:10.4269/ajtmh.2010.10-0082
PMCID: PMC2929047  PMID: 20810816
14.  The IFN-γ-Inducible GTPase, Irga6, Protects Mice against Toxoplasma gondii but Not against Plasmodium berghei and Some Other Intracellular Pathogens 
PLoS ONE  2011;6(6):e20568.
Clearance of infection with intracellular pathogens in mice involves interferon-regulated GTPases of the IRG protein family. Experiments with mice genetically deficient in members of this family such as Irgm1(LRG-47), Irgm3(IGTP), and Irgd(IRG-47) has revealed a critical role in microbial clearance, especially for Toxoplasma gondii. The in vivo role of another member of this family, Irga6 (IIGP, IIGP1) has been studied in less detail. We investigated the susceptibility of two independently generated mouse strains deficient in Irga6 to in vivo infection with T. gondii, Mycobacterium tuberculosis, Leishmania mexicana, L. major, Listeria monocytogenes, Anaplasma phagocytophilum and Plasmodium berghei. Compared with wild-type mice, mice deficient in Irga6 showed increased susceptibility to oral and intraperitoneal infection with T. gondii but not to infection with the other organisms. Surprisingly, infection of Irga6-deficient mice with the related apicomplexan parasite, P. berghei, did not result in increased replication in the liver stage and no Irga6 (or any other IRG protein) was detected at the parasitophorous vacuole membrane in IFN-γ-induced wild-type cells infected with P. berghei in vitro. Susceptibility to infection with T. gondii was associated with increased mortality and reduced time to death, increased numbers of inflammatory foci in the brains and elevated parasite loads in brains of infected Irga6-deficient mice. In vitro, Irga6-deficient macrophages and fibroblasts stimulated with IFN-γ were defective in controlling parasite replication. Taken together, our results implicate Irga6 in the control of infection with T. gondii and further highlight the importance of the IRG system for resistance to this pathogen.
doi:10.1371/journal.pone.0020568
PMCID: PMC3117789  PMID: 21698150
15.  Multiplex PCR Allows Rapid and Accurate Diagnosis of Bloodstream Infections in Newborns and Children with Suspected Sepsis▿†§ 
Journal of Clinical Microbiology  2011;49(6):2252-2258.
Sepsis is a major health problem in newborns and children. Early detection of pathogens allows initiation of appropriate antimicrobial therapy that strongly correlates with positive outcomes. Multiplex PCR has the potential to rapidly identify bloodstream infections, compensating for the loss of blood culture sensitivity. In an Italian pediatric hospital, multiplex PCR (the LightCycler SeptiFast test) was compared to routine blood culture with 1,673 samples obtained from 803 children with suspected sepsis; clinical and laboratory information was used to determine the patient infection status. Excluding results attributable to contaminants, SeptiFast showed a sensitivity of 85.0% (95% confidence interval [CI] = 78.7 to 89.7%) and a specificity of 93.5% (95% CI = 92.1 to 94.7%) compared to blood culture. The rate of positive results was significantly higher with SeptiFast (14.6%) than blood culture (10.3%) (P < 0.0001), and the overall positivity rate was 16.1% when the results of both tests were combined. Staphylococcus aureus (11.6%), coagulase-negative staphylococci (CoNS) (29.6%), Pseudomonas aeruginosa (16.5%), and Klebsiella spp. (10.1%) were the most frequently detected. SeptiFast identified 97 additional isolates that blood culture failed to detect (24.7% P. aeruginosa, 23.7% CoNS, 14.4% Klebsiella spp., 14.4% Candida spp.). Among specimens taken from patients receiving antibiotic therapy, we also observed a significantly higher rate of positivity of SeptiFast than blood culture (14.1% versus 6.5%, respectively; P < 0.0001). On the contrary, contaminants were significantly more frequent among blood cultures than SeptiFast (n = 97 [5.8%] versus n = 26 [1.6%]), respectively; P < 0.0001). SeptiFast served as a highly valuable adjunct to conventional blood culture in children, adding diagnostic value and shortening the time to result (TTR) to 6 h.
doi:10.1128/JCM.02460-10
PMCID: PMC3122766  PMID: 21471340
16.  Toxoplasma gondii infection and liver disease: a case-control study in a Northern Mexican population 
Parasites & Vectors  2011;4:75.
Background
Infection with the protozoan parasite Toxoplasma gondii may cause liver disease. However, the impact of the infection in patients suffering from liver disease is unknown. Therefore, through a case-control study design, 75 adult liver disease patients attending a public hospital in Durango City, Mexico, and 150 controls from the general population of the same region matched by gender, age, and residence were examined with enzyme-linked immunoassays for the presence of anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies. Socio-demographic, clinical and behavioral characteristics from the study subjects were obtained.
Results
Seroprevalence of anti-Toxoplasma IgG antibodies and IgG titers did not differ significantly in patients (10/75; 13.3%) and controls (16/150; 10.7%). Two (2.7%) patients and 5 (3.3%) controls had anti-Toxoplasma IgM antibodies (P = 0.57). Seropositivity to Toxoplasma did not show any association with the diagnosis of liver disease. In contrast, seropositivity to Toxoplasma in patients was associated with consumption of venison and quail meat. Toxoplasma seropositivity was more frequent in patients with reflex impairment (27.8%) than in patients without this impairment (8.8%) (P = 0.05). Multivariate analysis showed that Toxoplasma seropositivity in patients was associated with consumption of sheep meat (OR = 8.69; 95% CI: 1.02-73.71; P = 0.04) and rabbit meat (OR = 4.61; 95% CI: 1.06-19.98; P = 0.04).
Conclusions
Seropositivity to Toxoplasma was comparable among liver disease patients and controls. Further studies with larger sample sizes are needed to elucidate the association of Toxoplasma with liver disease. Consumption of venison, and rabbit, sheep, and quail meats may warrant further investigation.
doi:10.1186/1756-3305-4-75
PMCID: PMC3105944  PMID: 21569516
17.  Multicenter Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test as a Rapid Method for Detection of MRSA in Nasal Surveillance Swabs ▿  
Journal of Clinical Microbiology  2010;48(5):1661-1666.
The rate of methicillin-resistant Staphylococcus aureus (MRSA) infection continues to rise in many health care settings. Rapid detection of MRSA colonization followed by appropriate isolation can reduce transmission and infection. We compared the performance of the new Roche LightCycler MRSA advanced test to that of the BD GeneOhm MRSA test and culture. Double-headed swabs were used to collect anterior nasal specimens from each subject. For both tests, DNA was extracted and real-time PCR was performed according to manufacturer's instructions. For culture, one swab of the pair was plated directly to CHROMagar MRSA. The swab paired with the BD GeneOhm MRSA test was also placed into an enrichment broth and then plated to CHROMagar MRSA. Colonies resembling staphylococci were confirmed as S. aureus by standard methods. Discrepant specimens had further testing with additional attempts to grow MRSA as well as sample amplicon sequencing. Agreement between results for the two swabs was 99.3% for those with valid results. A total of 1,402 specimens were tested using direct culture detection of MRSA as the gold standard; 187 were culture positive for MRSA. The LightCycler MRSA advanced test had relative sensitivity and specificity of 95.2% (95% confidence interval [CI]: 91.1% to 97.8%) and 96.4% (95% CI: 95.2% to 97.4%), respectively. The BD GeneOhm assay had relative sensitivity and specificity of 95.7% (95% CI: 91.7% to 98.1%) and 91.7% (95% CI: 90.0% to 93.2%), respectively. Following discrepancy analysis, the relative sensitivities of the LightCycler MRSA advanced test and the BD GeneOhm MRSA assay were 92.2 and 93.2%, respectively; relative specificities were 98.9 and 94.2%, respectively. Specificity was significantly better (P < 0.001) with the LightCycler MRSA advanced test. The sensitivity of direct culture was 80.4%. The LightCycler MRSA advanced test is a useful tool for sensitive and rapid detection of MRSA nasal colonization.
doi:10.1128/JCM.00003-10
PMCID: PMC2863939  PMID: 20335423
18.  In Vitro and In Vivo Activities of 1-Hydroxy-2-Alkyl-4(1H)Quinolone Derivatives against Toxoplasma gondii▿  
1-Hydroxy-2-dodecyl-4(1H)quinolone (HDQ) was recently identified as a Toxoplasma gondii inhibitor. We describe here two novel 1-hydroxyquinolones, which displayed 50% inhibitory concentrations 10- and 5-fold lower than that of HDQ. In a mouse model of acute toxoplasmosis, these two compounds and HDQ reduced the percentages of infected peritoneal cells and decreased the parasite loads in lungs and livers. Compound B showed a tendency toward lowering parasite loads in brains in a mouse model of toxoplasmic encephalitis.
doi:10.1128/AAC.01001-09
PMCID: PMC2798484  PMID: 19884369
19.  Multiplex PCR To Diagnose Bloodstream Infections in Patients Admitted from the Emergency Department with Sepsis ▿  
Sepsis is caused by a heterogeneous group of infectious etiologies. Early diagnosis and the provision of appropriate antimicrobial therapy correlate with positive clinical outcomes. Current microbiological techniques are limited in their diagnostic capacities and timeliness. Multiplex PCR has the potential to rapidly identify bloodstream infections and fill this diagnostic gap. We identified patients from two large academic hospital emergency departments with suspected sepsis. The results of a multiplex PCR that could detect 25 bacterial and fungal pathogens were compared to those of blood culture. The results were analyzed with respect to the likelihood of infection, sepsis severity, the site of infection, and the effect of prior antibiotic therapy. We enrolled 306 subjects with suspected sepsis. Of these, 43 were later determined not to have infectious etiologies. Of the remaining 263 subjects, 70% had sepsis, 16% had severe sepsis, and 14% had septic shock. The majority had a definite infection (41.5%) or a probable infection (30.7%). Blood culture and PCR performed similarly with samples from patients with clinically defined infections (areas under the receiver operating characteristic curves, 0.64 and 0.60, respectively). However, blood culture identified more cases of septicemia than PCR among patients with an identified infectious etiology (66 and 46, respectively; P = 0.0004). The two tests performed similarly when the results were stratified by sepsis severity or infection site. Blood culture tended to detect infections more frequently among patients who had previously received antibiotics (P = 0.06). Conversely, PCR identified an additional 24 organisms that blood culture failed to detect. Real-time multiplex PCR has the potential to serve as an adjunct to conventional blood culture, adding diagnostic yield and shortening the time to pathogen identification.
doi:10.1128/JCM.01447-09
PMCID: PMC2812289  PMID: 19846634
20.  Interleukin (IL)-23 mediates Toxoplasma gondii–induced immunopathology in the gut via matrixmetalloproteinase-2 and IL-22 but independent of IL-17 
The Journal of Experimental Medicine  2009;206(13):3047-3059.
Peroral infection with Toxoplasma gondii leads to the development of small intestinal inflammation dependent on Th1 cytokines. The role of Th17 cells in ileitis is unknown. We report interleukin (IL)-23–mediated gelatinase A (matrixmetalloproteinase [MMP]-2) up-regulation in the ileum of infected mice. MMP-2 deficiency as well as therapeutic or prophylactic selective gelatinase blockage protected mice from the development of T. gondii–induced immunopathology. Moreover, IL-23–dependent up-regulation of IL-22 was essential for the development of ileitis, whereas IL-17 was down-regulated and dispensable. CD4+ T cells were the main source of IL-22 in the small intestinal lamina propria. Thus, IL-23 regulates small intestinal inflammation via IL-22 but independent of IL-17. Gelatinases may be useful targets for treatment of intestinal inflammation.
doi:10.1084/jem.20090900
PMCID: PMC2806449  PMID: 19995958
21.  A new two-phase dimeticone pediculicide shows high efficacy in a comparative bioassay 
BMC Dermatology  2009;9:12.
Background
Dimeticones kill head lice by physical means. Here we assessed in a comparative bioassay the ex vivo efficacy of "NYDA® sensitiv", a new two-phase dimeticone-based pediculicide similar to a product established on the market, but without fragrances.
Methods
We compared efficacy of the new product to a positive dimeticone control group, a sample of four other insecticidal and natural head lice products marketed in Germany, and an untreated control. In a bioassay, lice were exposed ex vivo to products and examined for activity for up to 24 hours, following a standard protocol.
Results
After 6 and 24 hours, 13.7 and 88.5% of untreated control lice did not show major vital signs. In contrast, no lice showed major vital signs 5 minutes after treatment with the new product or the control dimeticone group (NYDA®). This effect persisted at all observation points (100% efficacy). Efficacy of 0.5% permethrin (Infectopedicul®) ranged between 76 and 96% in evaluations between 5 min and 6 hours. All lice treated with a coconut-based compound (mosquito® Läuseshampoo) did not show major vital signs after 5 min, but mortality was only 58% after one hour. Pyrethrum extract (Goldgeist® forte) showed an efficacy of 22 - 52% between 5 min and 3 hours after treatment; after 6 hours, 76% of lice were judged dead. An oxyphthirine®-based compound (Liberalice DUO LP-PRO®) killed 22 - 54% of lice in the first 6 hours.
Conclusions
The two-phase dimeticone compound NYDA® sensitiv is highly efficacious. The removal of fragrances as compared to an established dimeticone product did not affect in vitro efficacy.
doi:10.1186/1471-5945-9-12
PMCID: PMC2800103  PMID: 20003435
22.  Aetiology of community-acquired, acute gastroenteritis in hospitalised adults: a prospective cohort study 
Background
The aetiology of severe gastroenteritis leading to hospitalisation in adults frequently remains unclear. Our objective was to study the causes and characteristics of community-acquired, acute gastroenteritis in adult hospitalized patients to support the clinical management of these patients.
Methods
From August 2005 to August 2007, we conducted a prospective cohort study among patients ≥18 y hospitalized with community-acquired gastroenteritis in a university hospital in Berlin, Germany. Stool specimens were examined for 26 gastrointestinal pathogens, supplemented by serologic tests for antibodies to Campylobacter spp., Yersinia spp., and Entamoeba histolytica. Patient data on demographics and clinical presentation were recorded and analyzed. Coexisting medical conditions were assessed using the Charlson Comorbidity Index score.
Results
Of 132 patients presenting with acute community-acquired gastroenteritis, 104 were included in the study. A non-infectious aetiology was diagnosed in 8 patients (8%). In 79 (82%) of the remaining 96 patients at least one microorganism was identified. Campylobacter spp. (35%) was detected most frequently, followed by norovirus (23%), Salmonella spp. (20%), and rotavirus (15%). In 46% of the patients with Campylobacter spp. infection, the diagnosis was made solely by serology. More than one pathogen was found in seventeen (22%) patients. Simultaneous infection was significantly more likely in patients with rotavirus and salmonella infections (RR 3.6; 95% CI: 1.8–7.4; RR 2.5; 95%CI: 1.2–5.5). Length of hospital stay (median: 5.5 days) was independent of the pathogen, but was associated with coexisting medical conditions (OR 4,8; 95%CI:2,0–11,6).
Conclusion
Known enteric pathogens were detected in 82% of adult patients who were hospitalized with acute gastroenteritis. We found that currently used culture-based methods may miss a substantial proportion of Campylobacter infections, and additional serological testing for Campylobacter should be considered. Viral infections emerged as an important cause of severe gastroenteritis in adults, and viral-bacterial co-infections in adults are probably underrecognized so far. The presence of coexisting medical conditions – but not the etiological agent – was a predictor for the duration of the hospital stay.
doi:10.1186/1471-2334-8-143
PMCID: PMC2596151  PMID: 18940017
23.  CD2 deficiency partially prevents small bowel inflammation and improves parasite control in murine Toxoplasma gondii infection 
AIM: To investigate whether bowel inflammation and/or parasite control is altered in the absence of the T cell adhesion molecule CD2.
METHODS: Wildtype (WT) and CD2 deficient (CD2-/-) mice were infected with 100 cysts of Toxoplasma gondii (T. gondii) (ME49) by gavage. On d 7 after infection mice were killed. Necrosis and the number of parasites/cm ileum were determined. Cytokine levels of stimulated cells as well as sera were evaluated. Secondly, survival of WT vs CD2-/- mice was analysed using Kaplan-Meier analysis.
RESULTS: CD2-/- mice survived longer than WT mice (mean: 23.5 vs 7.1 d, P = 0.001). Further, CD2-/- mice showed less weight loss and less ileal inflammation than WT mice at d 7 post infection. In addition, the number of parasites in the ileum was significantly lower in CD2-/- mice than in WT mice (88 ± 12 vs 349 ± 58 cm, P < 0.01). This was paralleled by lower production of IFN-γ and IL-6 from TLA-stimulated mLN cells and increased IFN-γ production by splenocytes.
CONCLUSION: CD2 deficient mice are more resistant to T. gondii infection than WT mice. In contrast to most current immunosuppressive or biological therapies CD2 deficiency reduces intestinal inflammation and at the same time helps to control infection.
doi:10.3748/wjg.v13.i31.4207
PMCID: PMC4250619  PMID: 17696249
CD2; IL-6; Ileitis; Inflammatory bowel disease; Interferon-γ; Toxoplasma gondii
24.  Shift Towards Pro-inflammatory Intestinal Bacteria Aggravates Acute Murine Colitis via Toll-like Receptors 2 and 4 
PLoS ONE  2007;2(7):e662.
Background
Gut bacteria trigger colitis in animal models and are suspected to aggravate inflammatory bowel diseases. We have recently reported that Escherichia coli accumulates in murine ileitis and exacerbates small intestinal inflammation via Toll-like receptor (TLR) signaling.
Methodology and Principal Findings
Because knowledge on shifts in the intestinal microflora during colitis is limited, we performed a global survey of the colon flora of C57BL/10 wild-type (wt), TLR2-/-, TLR4-/-, and TLR2/4-/- mice treated for seven days with 3.5% dextrane-sulfate-sodium (DSS). As compared to wt animals, TLR2-/-, TLR4-/-, and TLR2/4-/- mice displayed reduced macroscopic signs of acute colitis and the amelioration of inflammation was associated with reduced IFN-gamma levels in mesenteric lymph nodes, lower amounts of neutrophils, and less FOXP3-positive T-cells in the colon in situ. During acute colitis E. coli increased in wt and TLR-deficient mice (P<0.05), but the final numbers reached were significantly lower in TLR2-/-, TLR4-/- and TLR2/4-/- animals, as compared to wt controls (P<0.01). Concentrations of Bacteroides/ Prevotella spp., and enterococci did not increase during colitis, but their numbers were significantly reduced in the colon of DSS-treated TLR2/4-/- animals (P<0.01). Numbers of lactobacilli and clostridia remained unaffected by colitis, irrespective of the TLR-genotype of mice. Culture-independent molecular analyses confirmed the microflora shifts towards enterobacteria during colitis and showed that the gut flora composition was similar in both, healthy wt and TLR-deficient animals.
Conclusions and Significance
DSS-induced colitis is characterized by a shift in the intestinal microflora towards pro-inflammatory Gram-negative bacteria. Bacterial products exacerbate acute inflammation via TLR2- and TLR4-signaling and direct the recruitment of neutrophils and regulatory T-cells to intestinal sites. E. coli may serve as a biomarker for colitis severity and DSS-induced barrier damage seems to be a valuable model to further identify bacterial factors involved in maintaining intestinal homeostasis and to test therapeutic interventions based upon anti-TLR strategies.
doi:10.1371/journal.pone.0000662
PMCID: PMC1914380  PMID: 17653282
25.  Seroepidemiology of infection with Toxoplasma gondii in healthy blood donors of Durango, Mexico 
Background
Toxoplasma gondii (T. gondii) infection in blood donors could represent a risk for transmission in blood recipients. There is scarce information about the epidemiology of T. gondii infection in blood donors in Mexico. Therefore, we sought to determine the prevalence of T. gondii infection and associated socio-demographic and behavioral characteristics in a population of healthy blood donors of Durango City, Mexico.
Methods
Four hundred and thirty two blood donors in two public blood banks of Durango City, Mexico were examined for T. gondii infection between August to September 2006. Blood donors were tested for anti-T. gondii IgG and IgM antibodies by using enzyme-linked immunoassays (Diagnostic Automation Inc., Calabasas, CA, USA). Socio-demographic and behavioral characteristics from each participant were also obtained.
Results
Thirty two (7.4%) of 432 blood donors had IgG anti-T. gondii antibodies. Eight (1.9%) of them had also IgM anti-T. gondii antibodies. Multivariate analysis using logic regression showed that T. gondii infection was associated with the presence of cats at home (adjusted OR = 3.81; 95% CI: 1.45–10.01). The age group of 45–60 years showed a significantly higher frequency of T. gondii infection than the group of 25–34 years (p = 0.02). Blood donors without education had a significantly higher frequency of infection (15.8%) than those with 13–19 years of education (4.5%) (p = 0.04). Other characteristics of blood donors including male gender, consumption of undercooked meat or blood transfusion did not show an association with infection.
Conclusion
The prevalence of T. gondii infection in healthy blood donors of Durango City, Mexico is lower than those reported in blood donors of south and central Mexico, and is one of the lowest reported in blood donors worldwide. T. gondii infection in our blood donors was most likely acquired by contact with cats. Prevalence of infection increased with age and decreased with educational level.
doi:10.1186/1471-2334-7-75
PMCID: PMC1940003  PMID: 17629901

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