Isolation and characterization of viableToxoplasma gondiiisolates revealed possible high frequency of mixed infection in feral cats (Felis domesticus) from St Kitts, West Indies

Parasitology ◽  
2009 ◽  
Vol 136 (6) ◽  
pp. 589-594 ◽  
Author(s):  
J. P. DUBEY ◽  
L. MOURA ◽  
D. MAJUMDAR ◽  
N. SUNDAR ◽  
G. V. VELMURUGAN ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Mixed Infection ◽  
High Frequency ◽  
West Indies ◽  
Rflp Markers ◽  
Feral Cats ◽  
Clonal Type ◽  
Modified Agglutination Test ◽  
Isolation And Characterization ◽  
Pcr Rflp

SUMMARYCats are essential in the epidemiology ofToxoplasma gondiibecause they are the only hosts that can excrete the environmentally resistant oocysts in nature. Samples of serum, feces, and tissues from feral cats from St Kitts, West Indies were examined forT. gondiiinfection. Antibodies toT. gondiiwere assayed by the modified agglutination test, and found in 71 of 96 (73·9%) of cats with titres of 1:10 in six, 1: 20 in six,1:40 in seven,1: 80 in three, 1: 160 in 10, 1:320 in 13, 1:640 in nine, and 1:1,280 or higher in 17. Tissues of 10 cats were bio-assayed in mice.Toxoplasma gondiiwas isolated from tissues of 7 cats; from hearts of 6, from tongue of 5, and brains of 3 cats. All 7 isolates were avirulent for mice.Toxoplasma gondiioocysts were not found in the feces of 51 cats. Genotyping of these 7T. gondiiisolates by 10 multi-locus PCR-RFLP markers, including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker, Apico, revealed 4 genotypes, including clonal Type II, Type III and 2 unique genotypes. Five of the 7 cats had infection with 2 genotypes, indicating high frequency of mixed infection in the cat population on the St Kitts island.

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

Parasitology ◽  
2007 ◽  
Vol 134 (11) ◽  
pp. 1559-1565 ◽  
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.

scholarly journals Genetic characterization of Toxoplasma gondii isolates from eared doves (Zenaida auriculata) in Brazil

2014 ◽  
Vol 23 (4) ◽  
pp. 443-448 ◽  
Author(s):  
Luiz Daniel de Barros ◽  
Alessandra Taroda ◽  
Dauton Luiz Zulpo ◽  
Ivo Alexandre Leme da Cunha ◽  
Ana Sue Sammi ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Mouse Bioassay ◽  
Tissue Samples ◽  
Environmental Diversity ◽  
Clonal Type ◽  
Modified Agglutination Test ◽  
Pcr Rflp ◽  
Urban Rural ◽  
Toxoplasma Gondii Isolates

Eared doves (Zenaida auriculata), which are common in urban, rural and wild areas in many regions of Brazil, are frequently prey for domestic cats. Therefore Toxoplasma gondii isolates obtained from doves may reflect greater environmental diversity than those from other hosts. The aim of the present study was to evaluate T. gondii seroprevalence, isolate and genotype strains from Z. auriculata. Serum and tissue samples were collected from 206 doves for use in the modified agglutination test (MAT) and mouse bioassay. The prevalence of T. gondii antibodies in the doves was 22.3% (46/206), with titers ranging from 16 to 4096, and T. gondii strains were isolated from 12 of these doves. Five genotypes were detected by means of PCR-RFLP, including ToxoDB genotypes #1, #6, #17 and #65, and one genotype that had not previously been described (ToxoDB#182). This was the first report on isolation of T. gondii from Z. auriculata. This study confirmed the genetic diversity of T. gondii isolates and the existence of clonal type II (ToxoDB genotype #1) in Brazil.

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

Parasitology ◽  
2013 ◽  
Vol 140 (14) ◽  
pp. 1768-1776 ◽  
Author(s):  
A. BURRELLS ◽  
P. M. BARTLEY ◽  
I. A. ZIMMER ◽  
S. ROY ◽  
A. C. KITCHENER ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Rflp Markers ◽  
Type I ◽  
Type Ii ◽  
Red Foxes ◽  
Mammal Species ◽  
Tissue Samples ◽  
Pcr Rflp ◽  
Few Data ◽  
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.

Variation in Toxoplasma gondii seroprevalence: effects of site, sex, species and behaviour between insular and mainland macropods

2020 ◽  
Vol 47 (8) ◽  
pp. 540 ◽  
Author(s):  
Patrick L. Taggart ◽  
Bronwyn A. Fancourt ◽  
David Peacock ◽  
Charles G. B. Caraguel ◽  
Milton M. McAllister
Keyword(s):  
Toxoplasma Gondii ◽  
Wildlife Conservation ◽  
Road Mortality ◽  
High Seroprevalence ◽  
Feral Cats ◽  
Macropus Eugenii ◽  
Modified Agglutination Test ◽  
Latent Disease ◽  
Multiple Species ◽  
Indirect Impacts

Context Feral cats threaten wildlife conservation through a range of direct and indirect effects. However, most studies that have evaluated the impacts of feral cats on species of conservation significance have focussed on direct impacts such as predation; few studies have considered the indirect impacts of cat-borne disease. Toxoplasma gondii, a cat-borne parasite, causes both acute and latent disease in a range of wildlife species, and macropods are particularly susceptible. Kangaroo Island is Australia’s third largest island and supports a high density of feral cats and high seroprevalence of T. gondii in multiple species, relative to the mainland. This suggests that Kangaroo Island has a high environmental contamination with the parasite and a high risk of infection for other species. Aims We aimed to describe T. gondii seroprevalence in culled and road-killed macropods, so as to assess the effects of island versus mainland location, sex, species and behaviour. Methods Macropod sera were tested for T. gondii IgG antibodies using a commercially available modified agglutination test. Key results The seroprevalence of T. gondii in culled western grey kangaroos (Macropus fuliginosus) was significantly higher on the island (20%, 11/54 positive) than on the mainland (0%, 0/61 positive). There was no difference in T. gondii seroprevalence between culled and road-killed (21%, 21/102 positive) kangaroos from the island. The seroprevalence of T. gondii was significantly higher in female (32%, 12/38 positive) than in male (13%, 8/60 positive) kangaroos, but we observed no sex effect in tammar wallabies (Macropus eugenii), and no effect of species. Conclusions The higher T. gondii seroprevalence in insular macropods supports previous reports of higher T. gondii exposure in other Kangaroo Island fauna. The lack of difference in T. gondii seroprevalence between culled and road-killed kangaroos suggests that T. gondii-positive animals are not more vulnerable to road mortality, in contrast to that suggested previously. Implications Our findings suggest greater potential adverse conservation impacts owing to toxoplasmosis on the island than on the mainland. In light of a recent study demonstrating higher cat abundance on the island than on the mainland, the higher observed T. gondii seroprevalence in insular macropods is likely to be a consequence of higher cat density.

Toxoplasma gondii: Genotyping of strains from Brazilian AIDS patients with cerebral toxoplasmosis by multilocus PCR–RFLP markers

2008 ◽  
Vol 118 (2) ◽  
pp. 221-227 ◽  
Author(s):  
Isabelle M.R. Ferreira ◽  
José E. Vidal ◽  
Thais A. Costa-Silva ◽  
Cristina S. Meira ◽  
Roberto M. Hiramoto ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Rflp Markers ◽  
Cerebral Toxoplasmosis ◽  
Aids Patients ◽  
Pcr Rflp

scholarly journals SAG3 Toxoplasma gondii cloning reveals unexpected fivefold infection in the blood of feral cats in the Mexican Caribbean

2022 ◽  
Vol 18 (1) ◽  
Author(s):  
Luis Fernando Valenzuela-Moreno ◽  
Sara Teresa Méndez-Cruz ◽  
Claudia Patricia Rico-Torres ◽  
Carlos Cedillo-Peláez ◽  
Dolores Correa ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Toxoplasma Gondii ◽  
Bioinformatic Analysis ◽  
Restriction Pattern ◽  
Host Cells ◽  
Tropical Region ◽  
High Genetic Diversity ◽  
Feral Cats ◽  
Mexican Caribbean ◽  
Pcr Rflp

Abstract Background Currently, more than 300 genotypes of Toxoplasma gondii (T. gondii) have been described throughout the world, demonstrating its wide genetic diversity. The SAG3 locus is one of the genes included in the genotyping panel of this parasite. It is associated with its virulence since it participates during the invasion process of the host cells. Therefore, cloning, sequencing, and bioinformatic analysis were used to deepen the understanding of the SAG3 locus genetic diversity of T. gondii in blood samples from feral cats. Results Six different SAG3 sequences were detected, five of which were detected in one feline. Three sequences were first reported here; one of them was an intragenic recombinant. In the cladogram, four out of ten SAG3 sequences did not share nodes with others reported worldwide. Conclusions Cloning and sequencing of samples with more than one restriction pattern by PCR-RFLP were very helpful tools to demonstrate the presence of more than three genotypes of T. gondii in the blood of feral cats from southeastern Mexico. This suggests a potential mixed infection of multiple T. gondii strains and high genetic diversity of the parasites in felines in this tropical region of Mexico.

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

2012 ◽  
Vol 21 (1) ◽  
pp. 74-77 ◽  
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.

White-tailed deer (Odocoileus virginianus) are a reservoir of a diversity of Toxoplasma gondii strains in the USA and pose a risk to consumers of undercooked venison

Parasitology ◽  
2020 ◽  
Vol 147 (7) ◽  
pp. 775-781 ◽  
Author(s):  
J. P. Dubey ◽  
C. K. Cerqueira-Cézar ◽  
F. H. A. Murata ◽  
S. K. Verma ◽  
O. C. H. Kwok ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Cell Culture ◽  
Toxoplasma Gondii ◽  
Odocoileus Virginianus ◽  
Type Ii ◽  
Serum Samples ◽  
Modified Agglutination Test ◽  
Pcr Rflp ◽  
The Usa ◽  
Genotype 5

AbstractTo assess the role of white-tailed deer (Odocoileus virginianus, WTD) in the epidemiology of toxoplasmosis, we conducted a national survey of WTD across the USA for Toxoplasma gondii infection. To do this, we combined serology with parasite isolation to evaluate the prevalence and genetic diversity of T. gondii in this game species. From October 2012 to March 2019, serum and tissues were collected from 914 WTD across the USA. Serum samples were screened for antibodies to T. gondii, and then the tissues of seropositive WTD were bioassayed in mice. Antibodies were detected in 329 (36%) of 914 WTD tested by the modified agglutination test (positive reaction at 1:25 or higher). Viable T. gondii was isolated from the heart of 36 WTD from 11 states. Three of the 36 isolates were pathogenic but not highly virulent to outbred Swiss Webster mice and all 36 isolates could be propagated further in cell culture and were genotyped. For genotyping, DNA extracted from cell culture-derived tachyzoites was characterized by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using the genetic markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genotyping revealed seven ToxoDB PCR-RFLP genotypes, including 24 isolates for genotype #5 (haplogroup 12), four isolates for #2 (type III, haplogroup 3), three isolates for genotypes #1 (type II, haplogroup 2), two isolates for genotypes #3 (type II, haplogroup 2) and one isolate each for #39, #221 and #224. Genotype #5 was the most frequently isolated, accounting for 66.6% (24 of 36) of the isolates. Combining the 36 isolates from this study with previously reported 69 isolates from WTD, 15 genotypes have been identified. Among these, 50.4% (53/105) isolates belong to genotype #5. Our results indicate moderate genetic diversity of T. gondii in WTD. The results also indicate that undercooked venison should not be consumed by humans or fed to cats.

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

2019 ◽  
Vol 15 (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.

scholarly journals Fatal toxoplasmosis in an immunosuppressed domestic cat from Brazil caused by Toxoplasma gondii clonal type I

2017 ◽  
Vol 26 (2) ◽  
pp. 177-184 ◽  
Author(s):  
Hilda Fátima de Jesus Pena ◽  
Camila Mariellen Evangelista ◽  
Renata Assis Casagrande ◽  
Giovana Biezus ◽  
Claudia Salete Wisser ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Histopathological Examination ◽  
Leukemia Virus ◽  
Fatal Case ◽  
Domestic Cat ◽  
Feline Leukemia Virus ◽  
Type I ◽  
Clonal Type ◽  
Pcr Rflp ◽  
Pcr Targeting

Abstract The objective of the study was to report on a fatal case of feline toxoplasmosis with coinfection with the feline leukemia virus (FeLV). A domestic cat (Felis silvestris catus) presented intense dyspnea and died three days later. In the necropsy, the lungs were firm, without collapse and with many white areas; moderate lymphadenomegaly and splenomegaly were also observed. The histopathological examination showed severe necrotic interstitial bronchopneumonia and mild necrotic hepatitis, associated with intralesional cysts and tachyzoites of Toxoplasma gondii that were positive by anti-T. gondii immunohistochemical (IHC) evaluation. The bone marrow showed chronic myeloid leukemia and the neoplastic cells were positive by anti-FeLV IHC evaluation. DNA extracted from lungs was positive for T. gondii by PCR targeting REP-529. T. gondii was characterized by PCR-RFLP and by the microsatellites technique. ToxoDB-PCR-RFLP #10, i.e. the archetypal type I, was identified. Microsatellite analysis showed that the strain was a variant of type I with two atypical alleles. This was the first time that a T. gondii clonal type I genotype was correlated with a case of acute toxoplasmosis in a host in Brazil.

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