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.

Creole cattle from northwestern Mexico has high genetic diversity in the locus DRB3.2

2015 ◽  
Vol 57 ◽  
pp. 31-38
Author(s):  
I.G. Fernández ◽  
I. Leyva-Baca ◽  
F. Rodríguez-Almeida ◽  
R. Ulloa-Arvizu ◽  
J.G. Ríos-Ramírez ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Baja California ◽  
The State ◽  
Baja California Sur ◽  
High Genetic Diversity ◽  
La Paz ◽  
Separate Branch ◽  
Pcr Rflp ◽  
Creole Cattle ◽  
Northwestern Mexico

SummaryThe objective of this study was to determine the genetic diversity of creole cattle in northwestern Mexico using the BoLA-DRB3.2 locus of the Major Histocompatibility Complex (MHC). A total of 56 creole cattle were sampled from five communities; in the state of Chihuahua (Cerocahui, Guadalupe y Calvo and Cuauhtémoc) and in the state of Baja California Sur (La Paz and Mulegé). The BoLA-DRB3.2 locus was genotyped by PCR-RFLP assay. Thirty-nine alleles were identified, out of which 14 had not been previously reported. The average level of inbreeding in all populations analyzed wasFIS= 0.09 (P< 0.0001), but only two populations (Cerocahui and Guadalupe y Calvo) showed an excess of homozygotes (P< 0.05). The breed differentiation in all populations studied wasFSC= 0.068 (P< 0.0001). The smallest genetic distance was between La Paz and Mulegé (0.022); but Mulegé presented smaller distances (0.028–0.053) with the populations of La Paz (0.071–0.083) and with Chihuahua. Baja California Sur populations are grouped in a separate branch than Chihuahua populations. We conclude that creole cattle from Baja California Sur and Chihuahua show high genetic diversity in the locus BoLA-DRB3.2.

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 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.

Isolation and genotyping of Toxoplasma gondii in the Midwestern Brazil revealed high genetic diversity and new genotypes

Acta Tropica ◽  
2020 ◽  
Vol 212 ◽  
pp. 105681
Author(s):  
Rute Witter ◽  
Hilda Fátima Jesus Pena ◽  
Maerle Oliveira Maia ◽  
Aline Oliveira de Magalhães ◽  
Thaís Oliveira Morgado ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Toxoplasma Gondii ◽  
High Genetic Diversity

Resistance towards monensin is proposed to be acquired in a Toxoplasma gondii model by reduced invasion and egress activities, in addition to increased intracellular replication

Parasitology ◽  
2017 ◽  
Vol 145 (3) ◽  
pp. 313-325 ◽  
Author(s):  
AHMED THABET ◽  
JOHANNES SCHMIDT ◽  
SVEN BAUMANN ◽  
WALTHER HONSCHA ◽  
MARTIN VON BERGEN ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Toxoplasma Gondii ◽  
Metal Ion ◽  
Bioinformatic Analysis ◽  
Resistance Mechanisms ◽  
Host Cells ◽  
Intracellular Replication ◽  
Metal Ion Binding ◽  
Stable Isotope Labelling

SUMMARYMonensin (Mon) is an anticoccidial polyether ionophore widely used to control coccidiosis. The extensive use of polyether ionophores on poultry farms resulted in widespread resistance, but the underlying resistance mechanisms are unknown in detail. For analysing the mode of action by which resistance against polyether ionophores is obtained, we induced in vitro Mon resistance in Toxoplasma gondii-RH strain (MonR-RH) and compared it with the sensitive parental strain (Sen-RH). The proteome assessment of MonR-RH and Sen-RH strains was obtained after isotopic labelling using stable isotope labelling by amino acid in cell culture. Relative proteomic quantification between resistant and sensitive strains was performed using liquid chromatography-mass spectrometry/mass spectrometry. Overall, 1024 proteins were quantified and 52 proteins of them were regulated. The bioinformatic analysis revealed regulation of cytoskeletal and transmembrane proteins being involved in transport mechanisms, metal ion-binding and invasion. During invasion, actin and microneme protein 8 (MIC8) are seem to be important for conoid extrusion and forming moving junction with host cells, respectively. Actin was significantly upregulated, while MIC8 was downregulated, which indicate an invasion reduction in the resistant strain. Resistance against Mon is not a simple process but it involves reduced invasion and egress activity of T. gondii tachyzoites while intracellular replication is enhanced.

Genetic Diversity of Toxoplasma gondii Isolates in Egyptian Feral Cats Reveals New Genotypes

2010 ◽  
Vol 96 (6) ◽  
pp. 1112-1114 ◽  
Author(s):  
Y. M. Al-Kappany ◽  
C. Rajendran ◽  
S. A. Abu-Elwafa ◽  
M. Hilali ◽  
C. Su ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Toxoplasma Gondii ◽  
Feral Cats ◽  
Toxoplasma Gondii Isolates

scholarly journals Genetic Characterization of Toxoplasma gondii from Wild Rodents in Sichuan Province, Southwestern China

2019 ◽  
Author(s):  
XinLei WANG ◽  
Ling DONG ◽  
Li ZHANG ◽  
Yan LV ◽  
Qian LI ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Toxoplasma Gondii ◽  
Nested Pcr ◽  
Genetic Characterization ◽  
Sichuan Province ◽  
Southwestern China ◽  
Wild Rodents ◽  
New Genotype ◽  
Pcr Rflp

Background: Wild rodents are the intermediate hosts of Toxoplasma gondii. The distribution of genetic diversity of T. gondii in wild rodents is of importance to understand the transmission of this parasite. This study aimed to genetically characterize T. gondii isolates from wild rodents in Sichuan province, southwestern China in 2013. Methods: Genomic DNA was extracted from 10 g wild rodents’ brain samples. Semi-nested PCR and multilocous PCR-RFLP technology were performed to examine genetic diversity of T. gondii isolates as described previously. Results: Overall, 181 brain tissues of different wild rodents, including Eothenomys miletus (n=88), Crocidura attenuate (n=9), Rattus rattus sladeni (n=46), Mus musculus Linnaeus (n=6) and R. niviventer (n=32) were tested for T. gondii DNA, respectively. Six of them were positive for the T. gondii B1 gene by semi-nested PCR amplification, 4 showed complete genotyping results for all 11 polymorphic loci (SAG1, SAG2, alt. SAG2, SAG3, BTUB, GRA6, L358, PK1, C22-8, C29-2 and Apico) by PCR-RFLP, determined to represent a potential new genotype (http://toxodb.org/toxo/). Conclusion: These results documented genetic characterization of T. gondii in wild rodents from Sichuan province, and enriched the genetic diversity of T. gondii in China.

scholarly journals Genetic Diversity among Strains of Moraxella catarrhalis: Analysis Using Multiple DNA Probes and a Single-Locus PCR-Restriction Fragment Length Polymorphism Method

1998 ◽  
Vol 36 (7) ◽  
pp. 1977-1983 ◽  
Author(s):  
Elaine S. Walker ◽  
Robert A. Preston ◽  
J. Christopher Post ◽  
Garth D. Ehrlich ◽  
John H. Kalbfleisch ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Single Locus ◽  
High Genetic Diversity ◽  
Pcr Rflp ◽  
Restriction Fragment Length

Moraxella (Branhamella)catarrhalis, a causative agent of otitis media, sinusitis, and exacerbation of bronchitis, has acquired widespread ability to produce β-lactamase and can be nosocomially transmitted. The typing methods used in epidemiological analyses of M. catarrhalisare not optimal for genetic analyses. Two methods, a multiple-locus Southern blot (SB) method and a single-locus PCR-restriction fragment length polymorphism (RFLP) method, were developed and used to assess genetic diversity and potential clinical and geographic relationships in M. catarrhalis. Nine randomly cloned M. catarrhalis DNA fragments were used as probes of SBs containing DNA from 54 geographically and clinically diverse strains. For comparison, a PCR-RFLP method was developed as a quick, inexpensive, and discriminating alternative. A highly variable 3.7-kb genomic region (M46) was cloned and sequenced, and 3.5 kb of the cloned DNA was targeted for PCR amplification. DNAs from the 54 strains were subjected to PCR-RFLP. SB analysis distinguished all strains that had no apparent epidemiological linkage (40 of 54), and PCR-RFLP distinguished fewer strains (21 of 54). Epidemiologically linked strains appeared genetically identical by both methods. PCR-RFLP was compared to pulsed-field gel electrophoresis (PFGE) for 8 of the 54 strains and 23 additional strains. PCR-RFLP distinguished fewer strains than PFGE typing (16 of 31 versus 20 of 31 strains), but PCR-RFLP was more useful for inferring interstrain relatedness. Separate cluster analyses of multilocus SB and single locus PCR-RFLP data showed high genetic diversity within and across geographic locations and clinical presentations. The resultant dendrograms were not entirely concordant, but both methods often gave similar strain clusters at the terminal branches. High genetic diversity, nonconcordance of cluster analyses from different genetic loci, and shared genotypes among epidemiologically linked strains support a hypothesis of high recombination relative to spread of clones. Single-locus PCR-RFLP may be suitable for short-term epidemiological studies, but the SB data demonstrate that greater strain discrimination may be obtained by sampling variation at multiple genomic sites.

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.

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