Evidence of the three main clonalToxoplasma gondiilineages from wild mammalian carnivores in the UK

SUMMARYToxoplasma gondiiis a zoonotic pathogen defined by three main clonal lineages (types I, II, III), of which type II is most common in Europe. Very few data exist on the prevalence and genotypes ofT. gondiiin the UK. Wildlife can act as sentinel species forT. gondiigenotypes present in the environment, which may subsequently be transmitted to livestock and humans. DNA was extracted from tissue samples of wild British carnivores, including 99 ferrets, 83 red foxes, 70 polecats, 65 mink, 64 badgers and 9 stoats. Parasite DNA was detected using a nested ITS1 PCR specific forT. gondii, PCR positive samples were subsequently genotyped using five PCR–RFLP markers.Toxoplasma gondiiDNA was detected within all these mammal species and prevalence varied from 6·0 to 44·4% depending on the host. PCR–RFLP genotyping identified type II as the predominant lineage, but type III and type I alleles were also identified. No atypical or mixed genotypes were identified within these animals. This study demonstrates the presence of alleles for all three clonal lineages with potential for transmission to cats and livestock. This is the first DNA-based study ofT. gondiiprevalence and genotypes across a broad range of wild British carnivores.

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Isolation and genotyping of Toxoplasma gondii from pregnant dairy cows (Bos taurus) slaughtered

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612012000100016 ◽

2012 ◽

Vol 21 (1) ◽

pp. 74-77 ◽

Cited By ~ 12

Author(s):

Madlaine Frigo Silveira Barbosa de Macedo ◽

César Augusto Barbosa de Macedo ◽

Maria Paula de Carvalho Ewald ◽

Guilherme Felippelli Martins ◽

Dauton Luiz Zulpo ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Dairy Cows ◽

Blood Sample ◽

Bos Taurus ◽

Rflp Markers ◽

Human Consumption ◽

Mouse Bioassay ◽

Type Ii ◽

Tissue Samples ◽

Pcr Rflp

The current study aimed to evaluate serology, and isolate and genotype Toxoplasma gondii strains from pregnant dairy cows, slaughtered in an abattoir for human consumption, and their fetuses. Blood from 60 pregnant dairy cows and blood and tissue samples (brain, lung, heart, and liver) from their fetuses were collected and analyzed in a mouse bioassay. Antibodies against T. gondii were observed in 48.3% of cows and 3.7% of fetuses (IFAT, titers ≥ 50 for cows and 25 for fetuses were considered positive). Fourteen fetuses (23.3%) and six cows (10.0%) were identified as positive in the bioassay. T. gondii was isolated from a blood sample of a cow older than 4 years old in the 6th month of pregnancy, and from a blood sample of a fetus in the 6th month of gestation. These isolates were identified by polymerase chain reaction (PCR) as being of T. gondii and both strains showed type II alleles for all PCR-restriction fragment length polymorphism (PCR-RFLP) markers tested. T. gondii type II strain from cattle was isolated for the first time in Brazil. The current study also showed that transplacental transmission of T. gondii naturally occurs in dairy cows (23.3%) from Southern Brazil.

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Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran

BMC Veterinary Research ◽

10.1186/s12917-019-2176-2 ◽

2019 ◽

Vol 15 (1) ◽

Cited By ~ 1

Author(s):

Majid Khodaverdi ◽

Gholamreza Razmi

Keyword(s):

Toxoplasma Gondii ◽

Low Frequency ◽

Positive Sample ◽

Rflp Markers ◽

Type Ii ◽

Fecal Samples ◽

Specific Pcr ◽

Stray Cats ◽

Pcr Rflp ◽

Flotation Technique

Abstract Background Cats as a definitive host have an important role in the epidemiology of toxoplasmosis in humans and animals. The aim of the study was to determine the frequency of Toxoplasma gondii infection and isolate and identify the genotypes of T. gondii in stray cats in the Mashhad suburb. Methods From April 2016 to August 2017, 175 fecal samples from stray cats and 31 brain samples from cats killed in driving accidents were collected. The fecal samples were examined by fecal flotation technique and T. gondii-specific PCR. The brain samples were investigated by T. gondii-specific PCR and consequently examined by mice bioassay. The DNA of T. gondii isolated was genotyped using SAG1, SAG2, SAG3, BTUB and GRA6 as PCR-restriction fragment length polymorphism (PCR-RFLP) markers. Results In the present study, Toxoplasma-like oocysts were microscopically observed in 2.2% (4/175) fecal samples. The presence of Toxoplasma oocysts was confirmed in one microscopy-positive sample by PCR. In addition, T. gondii DNA was detected in 4% (7/175) microscopy-negative samples using PCR. T. gondii was isolated from one brain PCR-positive sample by mice bioassay. The isolate was avirulent and many T. gondii cysts were observed in mice brain. The isolate was successfully genotyped by PCR-RLFP analysis. The isolated genotyped was type II. Besides, eight Toxoplasma-positive fecal samples contained insufficient DNA and only amplified at SAG-3 locus in PCR. These samples were also showed type II pattern at this locus. Conclusions Parasitological and molecular results showed low frequency of Toxoplasma infection in the stray cats, and identified the genotype of T. gondii isolate as type II, for the first time in Mashhad area, Khorasan Razavi Province.

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Geographical patterns of Toxoplasma gondii genetic diversity revealed by multilocus PCR-RFLP genotyping

Parasitology ◽

10.1017/s0031182013001844 ◽

2013 ◽

Vol 141 (4) ◽

pp. 453-461 ◽

Cited By ~ 212

Author(s):

E. KEATS SHWAB ◽

XING-QUAN ZHU ◽

DEBASHREE MAJUMDAR ◽

HILDA F. J. PENA ◽

SOLANGE M. GENNARI ◽

...

Keyword(s):

Toxoplasma Gondii ◽

North America ◽

South America ◽

Type I ◽

Type Ii ◽

Distinct Population ◽

Geographical Patterns ◽

Pcr Rflp ◽

Population Structures ◽

Extensive Collection

SUMMARYIn recent years, an extensive collection of Toxoplasma gondii samples have been typed using a set of 10 PCR-RFLP genetic markers. Here we summarize the data reported until the end of 2012. A total of 1457 samples were typed into 189 genotypes. Overall, only a few genotypes dominate in the northern hemisphere, which is in stark contrast to the southern hemisphere where hundreds of genotypes coexist with none being notably dominant. PCR-RFLP genotype #1 (Type II clonal), #2 (Type III), #3 (Type II variant) and #10 (Type I) are identified globally. Genotypes #2 and #3 dominate in Africa, genotypes #9 (Chinese 1) and #10 are prevalent in Asia, genotypes #1, #2 and #3 are prevalent in Europe, genotypes #1, #2, #3, #4 and #5 dominate in North America (#4 and #5 are collectively known as Type 12). In Central and South America, there is no clear dominance of any genotype even though a few have relatively higher frequencies. Statistical analysis indicates significant differences among populations in Africa, Asia, Europe, North America, and Central and South America, with only Europe and North America exhibiting similar diversity. Collectively, the results revealed distinct population structures and geographical patterns of diversity in T. gondii.

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Comprehensive Characterization of Toxoplasma Acyl Coenzyme A-Binding Protein TgACBP2 and Its Critical Role in Parasite Cardiolipin Metabolism

mBio ◽

10.1128/mbio.01597-18 ◽

2018 ◽

Vol 9 (5) ◽

Cited By ~ 4

Author(s):

Yong Fu ◽

Xia Cui ◽

Sai Fan ◽

Jing Liu ◽

Xiao Zhang ◽

...

Keyword(s):

Fatty Acids ◽

Lipid Metabolism ◽

Toxoplasma Gondii ◽

Binding Protein ◽

Ankyrin Repeat ◽

Type I ◽

Type Ii ◽

Content Type ◽

High K ◽

Acyl Coenzyme A

ABSTRACT Acyl coenzyme A (CoA)-binding protein (ACBP) can bind acyl-CoAs with high specificity and affinity, thus playing multiple roles in cellular functions. Mitochondria of the apicomplexan parasite Toxoplasma gondii have emerged as key organelles for lipid metabolism and signaling transduction. However, the rationale for how this parasite utilizes acyl-CoA-binding protein to regulate mitochondrial lipid metabolism remains unclear. Here, we show that an ankyrin repeat-containing protein, TgACBP2, is localized to mitochondria and displays active acyl-CoA-binding activities. Dephosphorylation of TgACBP2 is associated with relocation from the plasma membrane to the mitochondria under conditions of regulation of environmental [K+]. Under high [K+] conditions, loss of ACBP2 induced mitochondrial dysfunction and apoptosis-like cell death. Disruption of ACBP2 caused growth and virulence defects in the type II strain but not in type I parasites. Interestingly, mitochondrial association factor-1 (MAF1)-mediated host mitochondrial association (HMA) restored the growth ability of ACBP2-deficient type II parasites. Lipidomics analysis indicated that ACBP2 plays key roles in the cardiolipin metabolism of type II parasites and that MAF1 expression complemented the lipid metabolism defects of ACBP2-deficient type II parasites. In addition, disruption of ACBP2 caused attenuated virulence of Prugniuad (Pru) parasites for mice. Taking the results collectively, these data indicate that ACBP2 is critical for the growth and virulence of type II parasites and for the growth of type I parasites under high [K+] conditions. IMPORTANCE Toxoplasma gondii is one of the most successful human parasites, infecting nearly one-third of the total world population. T. gondii tachyzoites residing within parasitophorous vacuoles (PVs) can acquire fatty acids both via salvage from host cells and via de novo synthesis pathways for membrane biogenesis. However, although fatty acid fluxes are known to exist in this parasite, how fatty acids flow through Toxoplasma lipid metabolic organelles, especially mitochondria, remains unknown. In this study, we demonstrated that Toxoplasma expresses an active ankyrin repeat containing protein TgACBP2 to coordinate cardiolipin metabolism. Specifically, HMA acquisition resulting from heterologous functional expression of MAF1 rescued growth and lipid metabolism defects in ACBP2-deficient type II parasites, manifesting the complementary role of host mitochondria in parasite cardiolipin metabolism. This work highlights the importance of TgACBP2 in parasite cardiolipin metabolism and provides evidence for metabolic association of host mitochondria with T. gondii.

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Muscle Biopsy Samples for Histochemical Processing: Alterations Induced by Storage

Veterinary Pathology ◽

10.1177/030098588802500111 ◽

1988 ◽

Vol 25 (1) ◽

pp. 77-82 ◽

Cited By ~ 6

Author(s):

K. G. Braund ◽

K. A. Amling

Keyword(s):

Skeletal Muscle ◽

Cell Morphology ◽

Type I ◽

Type Ii ◽

Tissue Samples ◽

Frozen Tissue ◽

Fresh Frozen ◽

Healthy Dogs ◽

Morphology And Morphometry ◽

Type Ii Fibers

Skeletal muscle samples from two healthy dogs were stored in ice at 0 C for up to 30 hours to examine the influence of time on cell morphology and morphometry. Cytochemical and histochemical properties of muscle to 18 hours were not markedly different from fresh frozen tissue. Samples stored to 30 hours were still satisfactory, despite a decline and unevenness in depth of staining. Morphometry from samples stored at 0 C for 6 hours or longer is not recommended, due to the statistically significant increase in diameter (from 21 to 25%) of type I and type II fibers.

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Comparative histochemical and morphometric analysis of muscle fibers of the psoas muscle in individuals of both genders with ageing

Vojnosanitetski pregled ◽

10.2298/vsp200713107a ◽

2020 ◽

pp. 107-107

Author(s):

Vladimir Antic ◽

Nenad Stojiljkovic ◽

Milorad Antic

Keyword(s):

Morphometric Analysis ◽

Statistical Significance ◽

Muscle Fibers ◽

Age Groups ◽

Psoas Muscle ◽

Iliopsoas Muscle ◽

Type I ◽

Type Ii ◽

Tissue Samples ◽

The Right

Background/Aim. In this paper, we analyzed type I and type II muscle fibers of the iliopsoas muscle in persons of both genders with ageing. The aim of this study was to detect the presence and distribution of types I and II muscle fibers in the human psoas muscle using the hematoxylin and eosin method in individuals of different ages and genders. Methods. The material consisted of tissue samples of the right iliopsoas muscle taken from 30 adult cadavers (18 males and 12 females), aged from 30 to 90 years, divided into three age groups. The material was obtained from the Institute of Forensic Medicine, Faculty of Medicine University of Nis. Results. The values of astereological parameters (area, perimeter and Feret's diameter) of type I and type II muscle fibers were higher in male cases, although without any statistical significance. Conclusion. Based on the histochemical and morphometric analysis, the conclusion was drawn that after 70 years of life there occurred a loss of type II muscle fibers, more conspicuous in female cases.

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Toxoplasma gondii and Neospora caninum infections in synanthropic rodents from Argentina

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612019009 ◽

2019 ◽

Vol 28 (1) ◽

pp. 113-118 ◽

Cited By ~ 1

Author(s):

Andrea Dellarupe ◽

Bruno Fitte ◽

Lais Pardini ◽

Lucía María Campero ◽

Mariana Bernstein ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Neospora Caninum ◽

Rattus Rattus ◽

Wide Distribution ◽

Type I ◽

Type Ii ◽

Caninum Infection ◽

Coccidian Parasites ◽

Rats And Mice ◽

Synanthropic Rodents

Abstract Toxoplasma gondii and Neospora caninum are closely related coccidian parasites (phylum Apicomplexa). This is the first study from urban synanthropic rodent species that involved serological and molecular diagnosis of T. gondii and N. caninum infection, and genotyping of T. gondii in Argentina. A total of 127 rodent samples were trap captured: Mus musculus (n = 78), Rattus norvegicus (n = 26) and Rattus rattus (n = 23). Antibodies against T. gondii and N. caninum were detected by IFAT in 32.8% (40/122) and 0.8% (1/122) of rodent samples, respectively, demonstrating contact with these protozoans. Additionally, T. gondii DNA was detected in 3.3% (4/123) of rodent central nervous system samples and 2 samples were genotyped by multilocus nPCR-RFLP. Neospora caninum DNA was not detected by PCR. The 2 genotyped samples were type III allele for all markers except for SAG-1 (type I for Rat1Arg and type II/III for Rat2Arg) and were identified as #48 and #2 (likely) according to the allele combinations reported on Toxo DB (Toxo-DB). The results of the present study revealed a wide distribution of T. gondii and less for N. caninum, in synanthropic rats and mice in the studied area.

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Molecular and biological characterization ofToxoplasma gondiiisolates from free-range chickens from Guyana, South America, identified several unique and common parasite genotypes

Parasitology ◽

10.1017/s0031182007003083 ◽

2007 ◽

Vol 134 (11) ◽

pp. 1559-1565 ◽

Cited By ~ 31

Author(s):

J. P. DUBEY ◽

L. APPLEWHAITE ◽

N. SUNDAR ◽

G. V. VELMURUGAN ◽

L. A. BANDINI ◽

...

Keyword(s):

Toxoplasma Gondii ◽

South America ◽

Rflp Markers ◽

Free Range ◽

Clonal Type ◽

Modified Agglutination Test ◽

Pcr Rflp ◽

Free Ranging ◽

Common Parasite

SUMMARYThe prevalence ofToxoplasma gondiiin free-ranging chickens (Gallus domesticus) is a good indicator of the prevalence ofT. gondiioocysts in the soil because chickens feed from the ground. The prevalence ofT. gondiiin 76 free-range chickens from Guyana, South America was determined. Antibodies toT. gondiiwere assayed by the modified agglutination test (MAT), and found in 50 (65·8%) of 76 chickens with titres of 1:5 in four, 1:10 in one, 1:20 in five, 1:40 in seven, 1:80 in six, 1:160 in eight, 1:320 in four, 1:640 or higher in 15. Hearts and brains of 26 chickens with titres of <1:5 were pooled in 5 batches and bioassayed in mice. Hearts and brains of 50 chickens with titres of 1:5 or higher were bioassayed in mice.Toxoplasma gondiiwas isolated by bioassay in mice from 35 chickens with MAT titres of 1:20 or higher. All mice inoculated with tissues of 30 infected chickens remained asymptomatic.Toxoplasma gondiiisolates from 35 chickens were genotyped using 11 PCR-RFLP markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, a new SAG2, and Apico. A total of 9 genotypes were identified, with 5 genotypes (nos 1, 4, 5, 6 and 7) unique to Guyana, 2 genotypes (nos 2 and 3) previously identified in chickens from Brazil, 1 genotype (no. 8) previously identified in chickens from Brazil, Costa Rica and Nicaragua, and 1 genotype (no. 9) belonging to the clonal type III lineage that exists globally. Infection with 2 genotypes was found from 1 chicken. This is the first report of genetic characterization ofT. gondiiisolates from any host from Guyana.

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Phosphoinositide-3-Kinase-Dependent, MyD88-Independent Induction of CC-Type Chemokines Characterizes the Macrophage Response to Toxoplasma gondii Strains with High Virulence

Infection and Immunity ◽

10.1128/iai.00821-07 ◽

2007 ◽

Vol 75 (12) ◽

pp. 5788-5797 ◽

Cited By ~ 13

Author(s):

Chiang W. Lee ◽

Woraporn Sukhumavasi ◽

Eric Y. Denkers

Keyword(s):

Toxoplasma Gondii ◽

Type I ◽

Type Ii ◽

Toll Like Receptor ◽

Innate And Adaptive Immunity ◽

Macrophage Response ◽

High Virulence ◽

Large Panel ◽

Phosphoinositide 3 Kinase ◽

Intracellular Infections

ABSTRACT Chemokines play an important role in inflammation and infection due to their ability to recruit cells of innate and adaptive immunity. Here we examined mouse macrophage chemokine responses during intracellular infections with high- and low-virulence Toxoplasma gondii strains. The high-virulence type I strain RH induced a large panel of CC-type chemokines, whereas responses elicited by strains PTG (type II) and M7741 (type III) were much weaker. Strikingly, the T. gondii-induced chemokine response occurred independently of signaling through the Toll-like receptor adaptor MyD88. Instead, production of chemokines during infection was heavily dependent upon phosphoinositide-3-kinase signaling pathways. Because infection with type I strains such as RH results in an uncontrolled proinflammatory cytokine response, we hypothesize that this virulence phenotype is a consequence of early strong induction of chemokines by type I, but not type II or III, Toxoplasma strains.

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