scholarly journals Detection and genetic characterisation of Toxoplasma gondii circulating in free-range chickens, pigs and seropositive pregnant women in Benue state, Nigeria

2021 ◽  
Vol 15 (6) ◽  
pp. e0009458
Author(s):  
Ifeoma N. Nzelu ◽  
Jacob K. P. Kwaga ◽  
Junaidu Kabir ◽  
Idris A. Lawal ◽  
Christy Beazley ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Toxoplasma Gondii ◽  
Pregnant Women ◽  
Quantitative Pcr ◽  
Human Consumption ◽  
Type I ◽  
Nested Polymerase Chain Reaction ◽  
Free Range ◽  
Animal Populations ◽  
Pcr Rflp

Toxoplasma gondii parasites present strong but geographically varied signatures of population structure. Populations sampled from Europe and North America have commonly been defined by over-representation of a small number of clonal types, in contrast to greater diversity in South America. The occurrence and extent of genetic diversity in African T. gondii populations remains understudied, undermining assessments of risk and transmission. The present study was designed to establish the occurrence, genotype and phylogeny of T. gondii in meat samples collected from livestock produced for human consumption (free-range chickens, n = 173; pigs, n = 211), comparing with T. gondii detected in blood samples collected from seropositive pregnant women (n = 91) in Benue state, Nigeria. The presence of T. gondii DNA was determined using a published nested polymerase chain reaction, targeting the 529 bp multicopy gene element. Samples with the highest parasite load (assessed using quantitative PCR) were selected for PCR-restriction fragment length polymorphism (PCR-RFLP) targeting the surface antigen 3 (SAG3), SAG2 (5’ and 3’), beta-tubulin (BTUB) and dense granule protein 6 (GRA6) loci, and the apicoplast genome (Apico). Toxoplasma gondii DNA was detected in all three of the populations sampled, presenting 30.6, 31.3 and 25.3% occurrence in free-range chickens, pigs and seropositive pregnant women, respectively. Quantitative-PCR indicated low parasite occurrence in most positive samples, limiting some further molecular analyses. PCR-RFLP results suggested that T. gondii circulating in the sampled populations presented with a type II genetic background, although all included a hybrid type I/II or II/III haplotype. Concatenation of aligned RFLP amplicon sequences revealed limited diversity with nine haplotypes and little indication of host species-specific or spatially distributed sub-populations. Samples collected from humans shared haplotypes with free-range chickens and/or pigs. Africa remains under-explored for T. gondii genetic diversity and this study provides the first detailed definition of haplotypes circulating in human and animal populations in Nigeria.

Genetic characterization of Toxoplasma gondii in Iranian HIV positive patients using multilocus nested-PCR-RFLP method

Parasitology ◽  
2019 ◽  
Vol 147 (3) ◽  
pp. 322-328
Author(s):  
Seyed Abdollah Hosseini ◽  
Mehdi Sharif ◽  
Shahabeddin Sarvi ◽  
Saeid Abediankenari ◽  
Mohammad Bagher Hashemi-Soteh ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hiv Positive ◽  
Type I ◽  
Serum Samples ◽  
Nested Polymerase Chain Reaction ◽  
Northern Iran ◽  
Type Iii ◽  
Genotype 2 ◽  
Pcr Rflp

AbstractThe aim of the present study was to investigate the prevalence and genotyping of Toxoplasma gondii in Iranian human immunodeficiency virus (HIV)-positive patients using multilocus-nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP). A total of 102 serum samples obtained from infected patients were collected from the laboratory centres in northern Iran. Anti-T. gondii antibodies and deoxyribonucleic acid (DNA) detection were accomplished by an enzyme-linked immunosorbent assay and PCR. The Mn-PCR-RFLP method was used for the genotyping of T. gondii. Overall, 68.6% (70/102) and 11.7% (12/102) of the individuals were tested positive for anti-T. gondii immunoglobulin G and T. gondii DNA, respectively. Complete genotyping was performed on 10/12 (83.3%) PCR-positive samples. Accordingly, the samples were classified as genotype #1 (type II clonal; n = 3, 30%), genotype #2 (type III clonal; n = 2, 20%), genotype #10 (type I clonal; n = 2, 20%), genotype #27 (type I variant; n = 1, 10%), genotype #35 (type I variant; n = 1, 10%) and genotype #48 (type III variant; n = 1, 10%). The results were indicative of the high frequency of the type I and type I variant of T. gondii strains in HIV-positive patients in northern Iran. Given the high prevalence of T. gondii and frequency of pathogenic types (pathogen in laboratory mice) in the patients, special measures should be taken to prevent the possible increased incidence of encephalitis by T. gondii.

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.

scholarly journals Molecular characterization of Toxoplasma gondii isolates from free-range chickens reveals new genotypes in Goiânia, Goiás, Brazil

2021 ◽  
Vol 30 (2) ◽  
Author(s):  
Hanstter Hallison Alves Rezende ◽  
Jaqueline Ataíde Silva Lima da Igreja ◽  
Antônio Roberto Gomes-Júnior ◽  
Jade de Oliveira Melo ◽  
João Luís Garcia ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Metropolitan Area ◽  
Seroprevalence Rate ◽  
Free Range ◽  
Low Genetic Diversity ◽  
Pcr Rflp ◽  
Peptic Digestion ◽  
Brain Cysts ◽  
Genotypic Characteristics

Abstract The aim of this study was to evaluate the genotypic characteristics of Toxoplasma gondii isolated from free-range chickens in the metropolitan area of Goiânia, Goiás, in Brazil’s central-west region. The seroprevalence rate was found to be 96%, according to an indirect hemagglutination assay. Brain and heart samples were processed by peptic digestion for a mice bioassay. The tissues were homogenized and the resulting samples were subjected to polymerase chain reaction (PCR), which revealed that 64% of them contained the parasite's DNA. The mice bioassay revealed 15 isolates, 8 of them tachyzoites isolates from the peritoneal lavage and 7 from brain cysts. T. gondii genotypes were determined through PCR-RFLP, using the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico and CS3. Three genotypes were identified, inclued ToxoDB #65, and the other two are not yet described in the literature. Hence, we conclude that the isolates obtained from the metropolitan area of Goiânia showed relatively low genetic diversity.

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 Detection of NaturalToxoplasma gondiiInfection in Chicken in Thika Region of Kenya Using Nested Polymerase Chain Reaction

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
John Mokua Mose ◽  
John Maina Kagira ◽  
Simon Muturi Karanja ◽  
Maina Ngotho ◽  
David Muchina Kamau ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxoplasma Gondii ◽  
Human Beings ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Free Range ◽  
Age Dependent ◽  
Polymerase Chain ◽  
The Difference

The detection ofToxoplasma gondiiin free-range chickens is a good indicator of possible risk to human beings. The aim of this study was to investigate the occurrence ofT. gondiiin free-range chicken using polymerase chain reaction (PCR). Brain samples from 105 free-range chickens from three administrative areas in Thika region, Kenya, were collected, DNA-extracted, and analyzed using PCR to detect presence ofT. gondii. The overall prevalence ofT. gondiiin all the three areas was 79.0% (95% CI: 70.0–86.4%) and the prevalence across the three areas was not significantly different (P=0.5088;χ2=1.354). Female chickens had higher (79.4%) prevalence than males (78.6%), although the difference was not significant (P=0.922,χ2= 0.01). However, chickens that were more than 2 years old had significantly (P=0.003;χ2= 11.87) higher prevalence compared to younger ones. The study indicates that there was a high occurrence ofT. gondiiinfection in free-range chickens from Thika region and that the infection rate is age dependent. Further studies should be carried out to determine the possible role of roaming chickens in the epidemiology of the disease among humans in the area.

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 Genotyping of Toxoplasma gondii isolates from naturally infected Gallus domesticus in Santa Catarina state, Brazil

2017 ◽  
Vol 69 (1) ◽  
pp. 139-145 ◽  
Author(s):  
N. Trevisani ◽  
L.D. Barros ◽  
A. Vieira-Neto ◽  
A.A. Sartor ◽  
A.P. Souza ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Toxoplasma Gondii ◽  
Immunofluorescence Assay ◽  
Gallus Domesticus ◽  
Mouse Bioassay ◽  
Type I ◽  
Santa Catarina ◽  
High Genetic Diversity ◽  
Clonal Population Structure ◽  
Santa Catarina State

ABSTRACT Toxoplasmosis is a widespread zoonosis that can infect warm-blooded animals including birds and humans, and chickens are considered to be indicators of environmental contamination. In Brazil, Toxoplasma gondii has a non-clonal population structure composed of three lineages (I, II, and III), presenting high recombination, and resulting in wide genetic diversity. This study aimed to genetically characterize T. gondii isolates from naturally infected chickens (Gallus domesticus) in Santa Catarina state, southern Brazil region. Sera from 133 free-range chickens were analyzed by an immunofluorescence assay (IFA) to detect IgG antibodies against T. gondii. Brain and heart from 30 positive animals, based on IFA (≥ 1:64), were used to isolate the parasite using a mouse bioassay. Strain genotyping was performed by PCR-RFLP using 12 genetic markers (SAG1, 5´-3´SAG2, alt. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3). The results were classified according to the genotypes based on the ToxoDB (http://toxodb.org/toxo/). Of 133 chicken sera analyzed, 84 (63.16%) were positive, with antibody titers ranging from 16 to 1024. Eleven isolates were obtained from mouse bioassay (Ck3, Ck32, Ck35, Ck56, Ck63, Ck89, Ck102, Ck103, Ck125, Ck127, and Ck128). Genotyping revealed six genotypes; three were classified as #26, #53, and #120, and three (NEO1, NEO2, and NEO3) were had not been previously described. No clonal lineages of type I, II, or III were identified. The present study confirms the high genetic diversity of T. gondii in Brazil.

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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