scholarly journals Determination of Chemical Inhibitor Efficiency against Intracellular Toxoplasma Gondii Growth Using a Luciferase-Based Growth Assay

Author(s):  
Melanie Key ◽  
Amy Bergmann ◽  
Chiara Micchelli ◽  
L. Brock Thornton ◽  
Sophie Millard ◽  
...  
2010 ◽  
Vol 73 (12) ◽  
pp. 2239-2243 ◽  
Author(s):  
SUSANA BAYARRI ◽  
MARÍA J. GRACIA ◽  
REGINA LÁZARO ◽  
CONSUELO PÉREZ-ARQUILLUÉ ◽  
MONTSERRAT BARBERÁN ◽  
...  

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii and distributed worldwide. Ingestion of viable cysts from infected raw or undercooked meat is an important route of horizontal transmission of the parasite to humans. Little information is available concerning the effect of commercial curing on cysts of T. gondii. This study is the first in which the influence of processing of cured ham on the viability of T. gondii has been evaluated, using bioassay to assess the risk of infection from eating this meat product. Naturally infected pigs were selected for the study, and a mouse concentration bioassay technique was used to demonstrate viable bradyzoites of T. gondii in porcine tissues and hams. No viable parasites were found in the final product (14 months of curing) based on results of the indirect immunofluorescence assay and histological and PCR analyses. Our results indicate that the consumption of hams cured as described here poses an insignificant risk of acquiring toxoplasmosis. However, additional studies are required to evaluate the safety of ham products cured under different conditions of curing time, salt, and nitrite concentration.


2002 ◽  
Vol 9 (2) ◽  
pp. 496-498
Author(s):  
Mardjan Arvand ◽  
Ilkay Kazak ◽  
Sergije Jovanovic ◽  
Hans-Dieter Foss ◽  
Oliver Liesenfeld

ABSTRACT We report on a young patient with chronic cervical lymphadenopathy and serological and histological evidence for infection with Bartonella henselae and Toxoplasma gondii. Serological follow-up studies, including testing for avidity of Toxoplasma-specific immunoglobulin G antibodies, assisted in the determination of the cause of the acute lymphadenitis. Our results suggest that the clinical symptoms were most likely due to cat scratch disease rather than to acute toxoplasmosis.


2014 ◽  
Vol 14 (5) ◽  
pp. 374-377 ◽  
Author(s):  
Byeong Yeal Jung ◽  
Eyerusalem Bizuneh Gebeyehu ◽  
Seung-Hun Lee ◽  
Min-Goo Seo ◽  
Jae-Won Byun ◽  
...  

2003 ◽  
Vol 47 (1) ◽  
pp. 309-316 ◽  
Author(s):  
Marc-Jan Gubbels ◽  
Catherine Li ◽  
Boris Striepen

ABSTRACT A high-throughput growth assay for the protozoan parasite Toxoplasma gondii was developed based on a highly fluorescent transgenic parasite line. These parasites are stably transfected with a tandem yellow fluorescent protein (YFP) and are 1,000 times more fluorescent than the wild type. Parasites were inoculated in optical-bottom 384-well culture plates containing a confluent monolayer of host cells, and growth was monitored by using a fluorescence plate reader. The signal was linearly correlated with parasite numbers over a wide array. Direct comparison of the YFP growth assay with the β-galactosidase growth assay by using parasites expressing both reporters demonstrated that the assays' sensitivities were comparable but that the accuracy of the YFP assay was higher, especially at higher numbers of parasites per well. Determination of the 50%-inhibitory concentrations of three known growth-inhibiting drugs (cytochalasin D, pyrimethamine, and clindamycin) resulted in values comparable to published data. The delayed parasite death kinetics of clindamycin could be measured without modification of the assay, making this assay very versatile. Additionally, the temperature-dependent effect of pyrimethamine was assayed in both wild-type and engineered drug-resistant parasites. Lastly, the development of mycophenolic acid resistance after transfection of a resistance gene in T. gondii was followed. In conclusion, the YFP growth assay limits pipetting steps to a minimum, is highly versatile and amendable to automation, and should enable rapid screening of compounds to fulfill the need for more efficient and less toxic antiparasitic drugs.


1998 ◽  
Vol 32 (2) ◽  
pp. 69-73 ◽  
Author(s):  
Hervé Pelloux ◽  
Emmanuelle Brun ◽  
Guy Vernet ◽  
Suzanne Marcillat ◽  
Michel Jolivet ◽  
...  

Author(s):  
Maria Aparecida Moraes Marciano ◽  
Rafaela Aparecida Silva ◽  
Maria Luisa Barbosa ◽  
Antônio Roberto Souza Ferreira ◽  
Vera Lucia Pereira-Chioccola

Author(s):  
James D. McKean ◽  
J. Beary ◽  
S. Brockus ◽  
Annette M. O'Connor ◽  
E. Zhou

1959 ◽  
Vol 37 (11) ◽  
pp. 1351-1360 ◽  
Author(s):  
C. G. Rogers ◽  
J. M. McLaughlan ◽  
D. G. Chapman

Bacteriological methods for the determination of protein quality were evaluated by comparison with protein efficiency ratio (P.E.R.) values determined by a standardized rat growth assay. Enzyme or acid hydrolyzates of foods were used as the source of amino acids with hydrolyzed whole egg powder as the reference standard. With Streptococcus faecalis A.T.C.C. 9790 autolysis occurred in media containing hydrolyzates of proteins deficient in lysine, and was largely responsible for results which did not agree with P.E.R. values. In methods employing Leuconostoc mesenteroides P-60 A.T.C.C. 8042, growth was influenced only by the most limiting amino acid relative to the requirements of the test organism.Results with enzyme hydrolyzates correlated poorly with P.E.R. values, whereas, with acid hydrolyzates, a good correlation was obtained for cereal proteins. A difference in amino acid requirements was largely responsible for the lack of agreement between the P.E.R. assay and methods employing L. mesenteroides, particularly for legumes and foods of animal origin. It was concluded that bacteriological assay methods which have been proposed for protein evaluation are unsatisfactory as screening procedures for the evaluation of protein in foods.


Parasitology ◽  
1984 ◽  
Vol 88 (3) ◽  
pp. 531-553 ◽  
Author(s):  
A. W. C. A. Cornelissen ◽  
J. P. Overdulve ◽  
M. Van Der Ploeg

SUMMARYDNA contents of individual stages of Isospora (Toxoplasma) gondii and other Eucoccida were measured after Feulgen-pararosaniline (SO2) staining either by direct microfluorometry or by scanning of microphotographic negatives. Frequency distributions were analysed using a computer program based on a mathematical model describing cell division. All stages of I. (T.) gondii, except fertilized macrogametes (2c), contained a haploid amount of DNA (1c), indicating that meiosis in I. (T.) gondii occurs during sporogony. Microgametes possessed 3·3% DNA in excess, presumably mitochondrial DNA. Nuclei of M2-and M3-merozoites differed in two characteristics: a small but distinct nucleolus was observed in almost 50% of the M2-merozoites but in none of the M3-merozoites; all M2 merozoites were strictly haploid, while all M3-merozoites were synthesizing DNA (17% above the haploid value). It may be concluded that all M2- and M3-merozoites are already sexually differentiated, i.e. are macro- and microgamontoblasts, respectively. DNA synthesis necessary for the development of the microgamont starts already in the microgamontoblast stage (M3-merozoite). M2-merozoites macrogametes, synthesize 11% extra DNA before fertilization, (after fertilization an extra amount of 12% of the diploid value was found), probably by amplification of genes for proteins which are needed for rapid maturation and later sporogony. Essentially parallel results have been found in Eimeria tenella and in crescent cystozoites of Sarcocystis cruzi. Absolute DNA values in representatives of the Eucoccida have been estimated as follows (10−15 g): I. (T.) gondii, 96; E. tenella and E. acervulina, both 75; S. cruzi, 216; Plasmodium berghei, 27. The value of the estimation of total haploid amounts as a tool in taxonomy of Eucoccida is discussed.


Sign in / Sign up

Export Citation Format

Share Document