The adaptive potential of a survival artist: characterization of the in vitro interactions of Toxoplasma gondii tachyzoites with di-cationic compounds in human fibroblast cell cultures

The impact of di-cationic pentamidine-analogues against Toxoplama gondii (Rh- and Me49-background) was investigated. The 72 h-growth assays showed that the arylimidamide DB750 inhibited the proliferation of tachyzoites of T. gondii Rh and T. gondii Me49 with an IC(50) of 0·11 and 0·13 μM, respectively. Pre-incubation of fibroblast monolayers with 1 μM DB750 for 12 h and subsequent culture in the absence of the drug also resulted in a pronounced inhibiton of parasite proliferation. However, upon 5-6 days of drug exposure, T. gondii tachyzoites adapted to the compound and resumed proliferation up to a concentration of 1·2 μM. Out of a set of 32 di-cationic compounds screened for in vitro activity against T. gondii, the arylimidamide DB745, exhibiting an IC(50) of 0·03 μM and favourable selective toxicity was chosen for further studies. DB745 also inhibited the proliferation of DB750-adapted T. gondii (IC(50)=0·07 μM). In contrast to DB750, DB745 also had a profound negative impact on extracellular non-adapted T. gondii tachyzoites, but not on DB750-adapted T. gondii. Adaptation of T. gondii to DB745 (up to a concentration of 0·46 μM) was much more difficult to achieve and feasible only over a period of 110 days. In cultures infected with DB750-adapted T. gondii seemingly intact parasites could occasionally be detected by TEM. This illustrates the astonishing capacity of T. gondii tachyzoites to adapt to environmental changes, at least under in vitro conditions, and suggests that DB745 could be an interesting drug candidate for further assessments in appropriate in vivo models.     

Evaluation of five antischizophrenic agents against Toxoplasma gondii in human cell cultures

An increasing interest in the association of the presence of antibodies to Toxoplasma gondii and the development of schizophrenia in patients has been generated over the last several years. Some antischizophrenia agents have been shown to have activity against T. gondii in cell culture assays and to ameliorate behavioral changes associated with chronic T. gondii infection in rats. In the present study, we examined the effects of commonly used antipsychotic and mood stabilizing agents (haloperidol, clozapine, fluphenazine, trifluoperazine, and thioridazine) for activity against developing tachyzoites of the RH strain of T. gondii in human fibroblast cell cultures. Neither haloperidol nor clozapine had a measurable effect. Fluphenazine had an IC(50) of 1.7 μM, thioridazine had an IC(50) of 1.2 μM, and trifluoperazine had an IC(50) of 3.8 μM. Our study demonstrates that some agents used to treat schizophrenia have the ability to inhibit T. gondii proliferation in cell culture.     

High prevalence and abundant atypical genotypes of Toxoplasma gondii isolated from lambs destined for human consumption in the USA

Little information is available on the presence of viable Toxoplasma gondii in tissues of lambs worldwide. The prevalence of T. gondii was determined in 383 lambs (<1 year old) from Maryland, Virginia and West Virginia, USA. Hearts of 383 lambs were obtained from a slaughter house on the day of killing. Blood removed from each heart was tested for antibodies to T. gondii by using the modified agglutination test (MAT). Sera were first screened using 1:25, 1:50, 1: 100 and 1:200 dilutions, and hearts were selected for bioassay for T. gondii. Antibodies (MAT, 1:25 or higher) to T. gondii were found in 104 (27.1%) of 383 lambs. Hearts of 68 seropositive lambs were used for isolation of viable T. gondii by bioassay in cats, mice or both. For bioassays in cats, the entire myocardium or 500g was chopped and fed to cats, one cat per heart and faeces of the recipient cats were examined for shedding of T. gondii oocysts. For bioassays in mice, 50g of the myocardium was digested in an acid pepsin solution and the digest inoculated into mice; the recipient mice were examined for T. gondii infection. In total, 53 isolates of T. gondii were obtained from 68 seropositive lambs. Genotyping of the 53 T. gondii isolates using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) revealed 57 strains with 15 genotypes. Four lambs had infections with two T. gondii genotypes. Twenty-six (45.6%) strains belong to the clonal Type II lineage (these strains can be further divided into two groups based on alleles at locus Apico). Eight (15.7%) strains belong to the Type III lineage. The remaining 22 strains were divided into 11 atypical genotypes. These results indicate high parasite prevalence and high genetic diversity of T. gondii in lambs, which has important implications in public health. We believe this is the first in-depth genetic analysis of T. gondii isolates from sheep in the USA.     

Toxoplasma gondii infection and schizophrenia: a case control study in a low Toxoplasma seroprevalence Mexican population

There are conflicting reports concerning the association of Toxoplasma gondii infection and schizophrenia in humans. Therefore, we determined such association in a Mexican population of Mestizo ethnicity. Through a case-control study design, 50 schizophrenic patients and 150 control subjects matched by gender, age, residence place, and ethnicity were examined with enzyme-linked immunoassays for the presence and levels of T. gondii IgG antibodies and for the presence of T. gondii IgM antibodies. Schizophrenic patients attended a public psychiatric hospital in Durango City, Mexico, and the control group consisted of individuals of the general population of the same city. Socio-demographic, clinical and behavioral characteristics from the study subjects were also obtained. Both the seroprevalence and the level of T.gondii IgG antibodies were higher in schizophrenic patients (10/50; 20%) than in control subjects (8/150; 5.3%) (OR=4.44; 95% CI: 1.49-13.37; P=0.003). The IgG T. gondii levels higher than 150 IU/ml were more frequently observed in patients than in controls (10% versus 2%, respectively; P=0.02). One (50%) of the two patients with recently diagnosed schizophrenia and none of the controls had T. gondii IgM antibodies (P=0.01). T. gondii seropositivity was significantly higher in patients with a history of cleaning cat excrement (P=0.005), and suffering from simple schizophrenia (ICD-10 classification: F20.6) (P=0.03) than patients without these characteristics. Toxoplasma seroprevalence was also significantly higher in patients with simple schizophrenia (F20.6) than in those with paranoid schizophrenia (F20.0) (P=0.02). This study provides elements to clarify the controversial information on the association of T. gondii infection and schizophrenia.     

Inhibition of Toxoplasma gondii and Plasmodium falciparum infections in vitro by NSC3852, a redox active antiproliferative and tumor cell differentiation agent

We searched the National Cancer Institute (NCI) compound library for structures related to the antitumor quinoline NSC3852 (5-nitroso-8-quinolinol) and used a computer algorithm to predict the antiprotozoan activity for each of 13 structures. Half of these compounds inhibited Toxoplasma gondii tachyzoite propagation in human fibroblasts at 相似文献   

Prevalence of antibodies to Toxoplasma gondii in raccoons (Procyon lotor) from an urban area of Northern Virginia

Raccoons (Procyon lotor) are intermediate hosts for Toxoplasma gondii, and clinical toxoplasmosis in raccoons has been reported. A 2-yr serological survey was conducted to determine the prevalence of antibodies to T. gondii in raccoons collected from Fairfax County, Virginia, a suburban/urban area outside Washington, D.C. Serum samples from 256 raccoons were examined for T. gondii antibodies at a 1:50 dilution using the modified direct agglutination test. Results indicated that 216 (84.4%) of the raccoons had been exposed to T. gondii. Our results indicate that raccoons in this area of Virginia are frequently exposed to T. gondii. Domestic cats were common in the study area and may have served as a source of oocysts for raccoons and the food items of raccoons.     

Seroprevalence of Toxoplasma gondii in elephants (Elephus maximus indicus) in Thailand

Serum samples from captive 156 elephants (Elephus maximus indicus) from Thailand were examined for antibodies to Toxoplasma gondii using the modified agglutination test (MAT) and the latex agglutination test (LAT). Antibodies to T. gondii were found in 45.5% of 156 elephants by MAT (> or = 1:50) and 25.6% of 156 elephants by LAT (> or = 1:64). This is the first report of T. gondii infection in E. maximus indicus from Asia.     

Synthesis and biological evaluation of 2-alkylaminoethyl-1,1-bisphosphonic acids against Trypanosoma cruzi and Toxoplasma gondii targeting farnesyl diphosphate synthase

The effect of a series of 2-alkylaminoethyl-1,1-bisphosphonic acids against proliferation of the clinically more relevant form of Trypanosoma cruzi, the etiologic agent of American trypanosomiasis (Chagas' disease), and against tachyzoites of Toxoplasma gondii has been studied. Most of these drugs exhibited an extremely potent inhibitory action against the intracellular form of T. cruzi, exhibiting IC(50) values at the low micromolar level. This cellular activity was associated with a strong inhibition of the enzymatic activity of T. cruzi farnesyl diphosphate synthase (TcFPPS), which constitutes a valid target for Chagas' disease chemotherapy. Compound 17 was an effective agent against amastigotes exhibiting an IC(50) value of 0.84 microM, while this compound showed an IC(50) value of 0.49 microM against the target enzyme TcFPPS. Interestingly, compound 19 was very effective against both T. cruzi and T. gondii exhibiting IC(50) values of 4.1 microM and 2.6 microM, respectively. In this case, 19 inhibited at least two different enzymes of T. cruzi (TcFPPS and solanesyl diphosphate synthase (TcSPPS); 1.01 microM and 0.25 microM, respectively), while it inhibited TgFPPS in T. gondii. In general, this family of drugs was less effective against the activity of T. cruzi SPPS and against T. gondii growth in vitro. As bisphosphonate-containing compounds are FDA-approved drugs for the treatment of bone resorption disorders, their potential low toxicity makes them good candidates to control tropical diseases.     

Prevalence and isolation of Toxoplasma gondii from white-tailed deer in Alabama

Nineteen white-tailed deer (Odocoileus virginianus) from 5 counties in Alabama were examined for infection with Toxoplasma gondii. Twenty-gram samples of heart tissue were bioassayed in mice, serum was examined for T. gondii antibodies using the direct agglutination test, and sections of heart muscle were examined histologically for tissue cysts. Toxoplasma gondii was isolated from 4 of 19 (21%) white-tailed deer hearts. Antibody titers of greater than or equal to 1:50 were found in sera from 7 of 16 (44%) white-tailed deer. Histological examinations of tissue sections from white-tailed deer hearts were negative for T. gondii.     

Toxoplasma gondii and Neospora caninum antibodies in dogs from Grenada, West Indies

Toxoplasma gondii and Neospora caninum are structurally similar parasites with many common hosts. The prevalence of antibodies to T. gondii and N. caninum was determined in sera from dogs in Grenada, West Indies. Using a modified agglutination test, antibodies to T. gondii were found in 52 (48.5%) of the 107 dogs, with titers of 1:25 in 17, 1:50 in 19, 1:100 in 7, 1:1,600 in 5, and 1:3,200 or higher in 4. Seroprevalence increased with age from 2.2% in dogs <6 mo old to 18.9% in dogs older than 2 yr, indicating postnatal transmission of T. gondii in this population of canines. There was no correlation between the health of the dogs and the seroprevalence or magnitude of the T. gondii titer. Antibodies to N. caninum were determined by the indirect immunofluorescent antibody test (IFAT). Two of the 107 dogs had N. caninum antibodies (IFAT titers 1:100 and 1:400); these dogs had T. gondii titers of 1:1,600 and 1:50, respectively. Results indicate that these 2 structurally similar protozoa are antigenically different.     

Seroprevalence of Toxoplasma gondii infection in chickens in Durango State, Mexico

Little is known concerning the seroprevalence of Toxoplasma gondii infection in chickens (Gallus domesticus) in Mexico. Antibodies to T. gondii were determined in 519 chickens in Durango, Mexico using the modified agglutination test (MAT). Two groups (A, B) of chickens were sampled. Group A chickens (n = 51) were raised in backyards in 7 municipalities in 3 geographical regions in Durango State. Group B chickens were raised in farms in the Mexican States of Sinaloa (n = 289) and Nayarit (n = 179) but slaughtered in 2 abattoirs in Durango City. Overall, antibodies to T. gondii were found in 36 (6.9%) of 519 chickens, with MAT titers of 1∶25 in 22, 1∶50 in 8, 1∶100 in 2, 1∶200 in 3, and 1∶400 in 1. Seroprevalence of T. gondii increased significantly with age and was significantly higher in Group A chickens than in Group B chickens. In Group A chickens, a 25.5% seroprevalence of T. gondii infection was found. Seropositive chickens were found in all 7 municipalities sampled. In Group B chickens, the seroprevalence of T. gondii infection was 4.9%. This is the first report of T. gondii infection in chickens in Durango State, Mexico.     

Seroprevalence of Toxoplasma gondii antibodies in domestic cats from Barcelona, Spain

Cats are important in the epidemiology of Toxoplasma gondii infection because they are the only hosts that can excrete the environmentally resistant oocysts. Antibodies to T. gondii were determined in serum samples from 220 domestic cats (Felis catus) from Barcelona, Spain, using the modified agglutination test (MAT). Antibodies to T. gondii were found in 99 (45%) of 220 cats, with MAT titers of 1:25 in 26, 1:50 in 57, and > or = 1:500 in 16 cats. Seropositivity (MAT 1:25 or more) was significantly higher in adult (> or = 1 yr old, 49.7% of 153) than in juvenile (< 1 yr old, 34.3% of 67) cats, in feral (51.9% of 131) than in domiciled (34.8% of 89) cats, and in cats living in a group (community) of more than 5 cats (50.7% of 142) than in cats living alone (28.0% of 50). These seropositive cats are likely to have already shed T. gondii oocysts in the environment around Barcelona.     

Novel triazine JPC-2067-B inhibits Toxoplasma gondii in vitro and in vivo

BACKGROUND AND METHODOLOGY: Toxoplasma gondii causes substantial morbidity, mortality, and costs for healthcare in the developed and developing world. Current medicines are not well tolerated and cause hypersensitivity reactions. The dihydrotriazine JPC-2067-B (4, 6-diamino-1, 2-dihydro-2, 2-dimethyl-1-(3'(2-chloro-, 4-trifluoromethoxyphenoxy)propyloxy)-1, 3, 5-triazine), which inhibits dihydrofolate reductase (DHFR), is highly effective against Plasmodium falciparum, Plasmodium vivax, and apicomplexans related to T. gondii. JPC-2067-B is the primary metabolite of the orally active biguanide JPC-2056 1-(3'-(2-chloro-4-trifluoromethoxyphenyloxy)propyl oxy)- 5-isopropylbiguanide, which is being advanced to clinical trials for malaria. Efficacy of the prodrug JPC-2056 and the active metabolite JPC-2067-B against T. gondii and T. gondii DHFR as well as toxicity toward mammalian cells were tested. PRINCIPAL FINDINGS AND CONCLUSIONS: Herein, we found that JPC-2067-B is highly effective against T. gondii. We demonstrate that JPC-2067-B inhibits T. gondii growth in culture (IC50 20 nM), inhibits the purified enzyme (IC50 6.5 nM), is more efficacious than pyrimethamine, and is cidal in vitro. JPC-2067-B administered parenterally and the orally administered pro-drug (JPC-2056) are also effective against T. gondii tachyzoites in vivo. A molecular model of T. gondii DHFR-TS complexed with JPC-2067-B was developed. We found that the three main parasite clonal types and isolates from South and Central America, the United States, Canada, China, and Sri Lanka have the same amino acid sequences preserving key binding sites for the triazine. SIGNIFICANCE: JPC-2056/JPC-2067-B have potential to be more effective and possibly less toxic treatments for toxoplasmosis than currently available medicines.     

First biologic and genetic characterization of Toxoplasma gondii isolates from chickens from Africa (Democratic Republic of Congo, Mali, Burkina Faso, and Kenya)

The prevalence of Toxoplasma gondii in free-ranging chickens (Gallus domesticus) is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. In the present study, prevalence of T. gondii in chickens from Democratic Republic of Congo, Mali, Burkina Faso, and Kenya is reported. The prevalence of T. gondii antibodies in sera of 50 free-range chickens from Congo was 50% based on the modified agglutination test (MAT); antibody titers were 1:5 in 7, 1:10 in 7, 1:20 in 6, 1:40 in 1, and 1:160 or more in 4 chickens. Hearts, pectoral muscles, and brains of 11 chickens with titers of 1:20 or more were bioassayed individually in mice; T. gondii was isolated from 9, from the hearts of 9, brains of 3, and muscles of 3 chickens. Tissues of each of the 14 chickens with titers of 1:5 or 1:10 were pooled and bioassayed in mice; T. gondii was isolated from 1 chicken with a titer of 1:10. Tissues from the remaining 25 seronegative chickens were pooled and fed to 1 T. gondii-free cat. Feces of the cat were examined for oocysts, but none was seen. The results indicate that T. gondii localizes in the hearts more often than in other tissues of naturally infected chickens. Genotyping of these 10 isolates using the SAG2 locus indicated that 8 were isolates were type III, 1 was type II, and 1 was type I. Two isolates (1 type I and 1 type III) were virulent for mice. Toxoplasma gondii was isolated by mouse bioassay from a pool of brains and hearts of 5 of 48 chickens from Mali and 1 of 40 chickens from Burkina Faso; all 6 isolates were avirulent for mice. Genetically, 4 isolates were type III and 2 were type II. Sera were not available from chickens from Mali and Burkina Faso. Toxoplasma gondii antibodies (MAT 100 or more) were found in 4 of 30 chickens from Kenya, and T. gondii was isolated from the brain of 1 of 4 seropositive chickens; this strain was avirulent for mice and was type II. This is the first report on isolation and genotyping of T. gondii from any source from these 4 countries in Africa.     

Seroprevalence of Toxoplasma gondii in harbor seals (Phoca vitulina) in southern Puget Sound, Washington.

As part of the Puget Sound Ambient Monitoring Program of the Washington Department of Fish and Wildlife, serum samples from 380 harbor seals (Phoca vitulina) were tested for antibodies to Toxoplasma gondii in the modified agglutination test (MAT) incorporating formalin-fixed tachyzoites and mercaptoethanol. Antibodies to T. gondii were found in 29 of 380 (7.6%) seals with titers of 1:25 in 13, 1:50 in 14, and > or = 1:500 in 2 seals. Results indicate natural exposure of these wild marine mammals to T. gondii oocysts.     

Tissue distribution and molecular characterization of chicken isolates of Toxoplasma gondii from Peru

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii antibodies in sera of 50 free-range chickens (Gallus domesticus) from Peru was 26% on the basis of the modified agglutination test (MAT). Hearts, pectoral muscles, and brains of seropositive (MAT > or =1:5) chickens were bioassayed individually in mice. Tissues from the remaining 37 seronegative chickens were pooled and fed to 2 T. gondii-free cats. Feces of cats were examined for oocysts; they did not shed oocysts. Toxoplasma gondii was isolated from the hearts of 10 seropositive chickens but not from their brains and pectoral muscles. Genotyping of these isolates using the SAG2 locus indicated that 7 isolates were type I and 3 were type III. Six of the 7 type-I isolates were avirulent for mice, which was unusual because type-I isolates are considered virulent for mice. The T. gondii isolates were from chickens from different properties that were at least 200 m apart. Thus, each isolate is likely to be different. This is the first report of isolation of T. gondii from chickens from Peru.     

Seroprevalence of Toxoplasma gondii in pigs from Vietnam

Pigs are considered an important source of Toxoplasma gondii infection for humans. Antibodies to T. gondii were determined in serum samples from 587 pigs from Vietnam using the modified agglutination test (MAT) and found in 160 of 587 (27.2%) pigs, with MAT titers of 1:25 in 32 pigs, 1:50 in 34 pigs, 1:100 in 33 pigs, 1:200 in 24 pigs, 1:400 in 21 pigs, 1:800 in 14 pigs, and 1:3,200 in 2 pigs. Antibodies (MAT 1:20 or higher) were found in 75 of 325 (23%) finishers, 63 of 207 (32.3%) sows, and 22 of 55 (40%) boars. Results indicate high prevalence of T. gondii infection in pigs in Vietnam. This is the first report of prevalence of T. gondii in pigs from Vietnam.     

Seroprevalence of Toxoplasma gondii infection in dogs in Jiangsu Province, eastern China

There is a lack of epidemiological data on Toxoplasma gondii infection in dogs from eastern China. In the present study, serum samples from 288 dogs were collected from Xuzhou, Huaiyin, and Yianchen in Jiangsu Province, eastern China, in August 2010; T. gondii antibodies were assayed by a modified agglutination test (MAT). Antibodies to T. gondii were found in 62 of 288 (21.5%) with MAT titers of 1∶25 in 21 dogs, 1∶50 in 15, 1∶100 in 11, 1∶200 in 6, and 1∶400 and above in 3 dogs. The seroprevalence in ≥3-yr-old dogs was significantly higher (P < 0.05) than that in dogs less than 3-yr-old. The results of the present study indicated that infection with T. gondii in household dogs in Jiangsu Province is a public health concern.     

Seroprevalence of Toxoplasma gondii in Rocky Mountain bighorn sheep (Ovis canadensis)

Serum samples from 697 Rocky Mountain bighorn sheep (Ovis canadensis) from North America were examined for antibodies to Toxoplasma gondii by the modified agglutination test incorporating mercaptoethanol and formalin-fixed tachyzoites. Antibodies to T. gondii were found in 25 of 697 (3.6%) sheep in titers of 1:25 (8 sheep), 1:50 (4 sheep), 1:100 (7 sheep), 1:200 (1 sheep), 1:400 (1 sheep), 1:800 (1 sheep), and 1:1,600 (3 sheep). This is the first record of T. gondii exposure in bighorn sheep.