Correction to: Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks.
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SUBMITTER: Fernandez-Escobar M
PROVIDER: S-EPMC7640693 | biostudies-literature | 2020 Nov
REPOSITORIES: biostudies-literature
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Correction to: Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks.
Fernández-Escobar Mercedes M Calero-Bernal Rafael R Benavides Julio J Regidor-Cerrillo Javier J Guerrero-Molina María Cristina MC Gutiérrez-Expósito Daniel D Collantes-Fernández Esther E Ortega-Mora Luis Miguel LM
Parasites & vectors 20201103 1
An amendment to this paper has been published and can be accessed via the original article. ...[more]
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Project description:BackgroundToxoplasma gondii is a major cause of abortion in small ruminants and presents a zoonotic risk when undercooked meat containing cysts is consumed. The aim of the present study was to investigate the genetic diversity among the T. gondii strains circulating in ovine livestock in Spain.MethodsSelected samples collected from abortion outbreaks due to toxoplasmosis (n = 31) and from chronically infected adult sheep at slaughterhouses (n = 50) in different Spanish regions were bioassayed in mice, aiming at parasite isolation. In addition, all original clinical samples and the resulting isolates were genotyped by multi-nested PCR-RFLP analysis of 11 molecular markers and by PCR-DNA sequencing of portions of the SAG3, GRA6 and GRA7 genes.ResultsAs a result, 30 isolates were obtained from 9 Spanish regions: 10 isolates from abortion-derived samples and 20 isolates from adult myocardial tissues. Overall, 3 genotypes were found: ToxoDB#3 (type II PRU variant) in 90% (27/30) of isolates, ToxoDB#2 (clonal type III) in 6.7% (2/30), and ToxoDB#1 (clonal type II) in 3.3% (1/30). When T. gondii-positive tissue samples (n = 151) were directly subjected to RFLP genotyping, complete restriction profiles were obtained for 33% of samples, and up to 98% of the specimens belonged to the type II PRU variant. A foetal brain showed a clonal type II pattern, and four specimens showed unexpected type I alleles at the SAG3 marker, including two foetal brains that showed I + II alleles as co-infection events. Amplicons of SAG3, GRA6 and GRA7 obtained from isolates and clinical samples were subjected to sequencing, allowing us to confirm RFLP results and to detect different single-nucleotide polymorphisms.ConclusionsThe present study informed the existence of a predominant type II PRU variant genotype (ToxoDB#3) infecting domestic sheep in Spain, in both abortion cases and chronic infections in adults, coexisting with other clonal (ToxoDB#1 and ToxoDB#2), much less frequent genotypes, as well as polymorphic strains as revealed by clinical sample genotyping. The use of multilocus sequence typing aided in accurately estimating T. gondii intragenotype diversity.
| S-EPMC7404076 | biostudies-literature
Project description:Many cases of Toxoplasma gondii infection have been reported worldwide in non-human primates (NHPs), especially in captive New World monkeys. However, few studies on toxoplasmosis in Old World monkeys have been conducted. In this study, serological and molecular biological analyses were carried out to look for T. gondii antibodies and T. gondii infection in 13 NHPs from China. T. gondii infection was confirmed in 8 NHP cases. T. gondii antibodies were detected in 1/5 New World monkeys and in 4/7 Old World monkeys. T. gondii DNA was detected in 3/5 New World monkeys and 5/7 Old World monkeys. The one ring-tailed lemur was negative for both antibodies and DNA of T. gondii. The most common clinical manifestations of T. gondii infection were malaise, poor appetite, emaciation, and foamy nasal discharge. The most common histopathological findings were interstitial pneumonia, necrotic hepatitis, necrotizing myocarditis, lymphadenitis, and necrotic splenitis. One viable T. gondii strain was successfully isolated from the myocardium of a patas monkey (Erythrocebus patas) by bioassay in mice. T. gondii tachyzoites were obtained from cell cultures and were designated as TgMonkeyCHn2. The genotype of this strain belongs to ToxoDB genotype #9, and the allele of ROP18/ROP5 gene was 3/6. TgMonkeyCHn2 tachyzoites were avirulent in Swiss mice. To our knowledge, this is the first report of fatal toxoplasmosis in a patas monkey. T. gondii infection in patas monkeys may indicate environmental contamination by oocysts. The patas monkey is a new host record for T. gondii.
| S-EPMC10454590 | biostudies-literature
Project description:BackgroundToxoplasmosis is a widespread zoonosis caused by the intracellular protozoan parasite Toxoplasma gondii. Limited epidemiological information is available about the prevalence of T. gondii in sheep in Romania, and a high incidence would have implications for both the economy and public health. To our knowledge, no studies are available about the T. gondii strains circulating in lambs. The objective of this study was to assess the prevalence of T. gondii in sheep (serology), lambs (serology, bioassay, PCR) and sheep abortions (PCR) in Romania. Moreover, the study aimed to perform the genetic characterization of T. gondii isolates from lambs.MethodsSerum samples collected from 2650 sheep (2067 adults and 583 lambs) were tested for anti-T. gondii antibodies (IgG) using a commercial ELISA kit. Likewise, 328 pairs of diaphragmatic muscle-serum samples were collected from lambs aged between 2 and 4 months. Lamb serum samples were analyzed using MAT for anti-T. gondii antibody detection. The diaphragm tissue samples from MAT-positive lambs (at a dilution ≥ 1:25) were bioassayed in mice. The T. gondii strains were genotyped using 15 microsatellites markers. Additionally, brain and heart samples from 76 sheep abortions were analyzed for T. gondii DNA by polymerase chain reaction (PCR) targeting the 529-bp repeat region (REP529).ResultsThe results showed that more than half of the tested sheep were T. gondii seropositive (53.5%). The seroprevalence was significantly higher in adults (61.1%) than in lambs (26.4%). The seroprevalence of T. gondii infection in slaughtered lambs, by MAT, was 37.5% (123/328). There were bioassayed in mice 56 diaphragmatic tissues from 123 seropositive lambs. Toxoplasma gondii strains were isolated from 18 (32.1%) lambs intended for human consumption. All T. gondii strains were confirmed by PCR. Six strains were genotyped using 15 microsatellite markers and belonged to genotype II. Toxoplasma gondii DNA was detected in 11.8% (9/76) of sheep abortions.ConclusionsThe present study showed the presence of T. gondii in sheep in all the regions considered in the study. The high prevalence of T. gondii infection in sheep and lambs, demonstrated by serology, molecular analysis and bioassay, highlighted that there is an important risk of human infection in consuming raw or undercooked sheep/lamb meat.
| S-EPMC9872358 | biostudies-literature
Project description:BackgroundThe consumption of uncooked or undercooked food from infected intermediate hosts can result in Toxoplasma gondii infection in humans. However, few studies have investigated the genetic diversity of this protozoan parasite in Iran. The aim of the present study was to genetically characterize isolates of T. gondii from intermediate host animals in Mazandaran Province, Iran.MethodsBlood and heart tissue samples were collected from 204 ruminants, and brain tissue was collected from 335 birds. The prevalence of T. gondii infection in these samples was determined serologically using the modified agglutination test and by conventional PCR assays. Those PCR samples positive for T. gondii DNA and 13 DNA samples extracted from aborted fetuses in a previous study were genotyped with 12 genetic markers using the multilocus-nested PCR-restriction fragment length polymorphism (Mn-PCR-RFLP) technique.ResultsAntibodies for parasites were found in 35.7% of the ruminant (39.1% of sheep and 26.4% of goats) samples and in 51.3% of the bird (100% of geese, 52.3% of free-range chickens and 46% of ducks) samples. Molecular detection by PCR of the repetitive 529-bp DNA fragment revealed contamination of 13.2% of ruminant (14.6% of sheep and 9.4% of goats) samples and of 9.6% of bird (11.1% of chickens, 5.7% of ducks and 0% of geese samples). The results from 30 DNA samples revealed five ToxoDB (genome database for the genus Toxoplasma) PCR-RFLP genotypes, including #1 (Type II), #2 (Type III), #10 (Type I), #27 and #48, with genotype #1 the most predominant.ConclusionsAs evidenced by the results of this study, ruminants and birds are infected with T. gondii in the region, suggesting that they are likely to be involved in the transmission of T. gondii to humans through meat consumption. The identification of different genotypes may suggest a higher genetic diversity of this parasite in Mazandaran, reflecting local environmental contamination. These results have important public health implications for the prevention and control strategies of infection.
| S-EPMC9277799 | biostudies-literature
Project description:BackgroundEpidemiological investigations on Toxoplasma gondii infection have found a significant association between human toxoplasmosis and consumption of raw or undercooked meat. The present study aimed to characterize genotypes of T. gondii in 20 cattle, 40 sheep, 15 goats and 16 pigs from the North of Portugal.MethodsNested PCR amplified the surface antigen 2 (SAG2) gene. Sequencing analysis was performed in order to assess the prevalence of SAG2 type strains (I, II and III).ResultsThree and 4 strains of SAG2 type II were identified in heart samples of cattle and sheep, respectively. Three SAG2 type II strains were detected in brain, diaphragm and heart of 3 pigs. Three strains detected in heart samples of 3 goats belonged to SAG2 types I or II; with the same result being observed in heart samples from 2 sheep and in 2 brain and 1 heart samples from 3 pigs.ConclusionSAG2 type II has been shown for the first time to infect cattle in North of Portugal. In addition, SAG2 type II has also been confirmed as the predominant strain in sheep and pigs in the same region. This is the first molecular report of T. gondii in goats from Portugal.
| S-EPMC4662747 | biostudies-literature
Project description:Cutaneous toxoplasmosis is a rare manifestation. This study represents a case report of an immunosuppressed dog that developed nodular dermal lesions caused by Toxoplasma gondii. The isolate (TgDgBr20) was characterized as mouse virulent and was genotyped as type BrI (ToxoDB genotype 6) using PCR-restriction fragment length polymorphism (RFLP) and as Africa 1 through microsatellite analysis.
| S-EPMC4313287 | biostudies-literature
Project description:BACKGROUND: Toxoplasma gondii is an obligate intracellular protozoan parasite that infects humans and a broad spectrum of warm-blooded vertebrates. The present study was undertaken with the objectives of isolation and determining the genotypes of T. gondii strains from sheep and goats slaughtered in East and West Shewa Zones of Oromia Regional State, Central Ethiopia. METHODS: Hearts of 47 sheep and 44 goats that were seropositive in the Direct Agglutination Test (DAT) were bioassayed in mice. A multiplex PCR assay with 15 microsatellite markers was employed for genotyping of T. gondii isolates from sheep and goats. RESULTS: Viable T. gondii were isolated from 47 (51.65%) animals, 27 sheep and 20 goats. Most isolates caused sub-clinical infections in mice, however, 2 sheep and 1 goat isolates were mouse-virulent, killing mice between 19-27 days post-inoculation. The success of T. gondii isolation in mice increased significantly (P?=?0.0001) with higher DAT antibody titers in sheep and goats. Genotyping revealed that 29 (87.88%) of the 33 isolates were Type II, 3 (9.09%) were Type III and 1 (3.03%) was atypical. Three strains (one type II, one type III, and the atypical genotype) were virulent for mice. CONCLUSIONS: T. gondii tissue cysts in sheep and goats slaughtered for human consumption are widespread. This is the first report on isolation and genotyping of T. gondii from sheep and goats of Ethiopia.
| S-EPMC4161867 | biostudies-other
Project description:The present study aimed to isolate and perform molecular and phenotypic characterization of Toxoplasma gondii strains infecting Iberian pigs bred under semi-free conditions and destined for human consumption. Blood and heart tissue samples from 361 fattening pigs from 10 various herds selected in the main areas of Iberian pig production were collected at a slaughterhouse; the sera were tested for anti-T. gondii antibodies using a commercial indirect ELISA kit, and a mouse bioassay was carried out using heart muscle of seropositive individual representatives from each geographical location. Seventy-nine (21.9%) of the 361 animals tested positive for anti-T. gondii antibodies according to the serology test. Fifteen samples of myocardial tissue were subjected to bioassay and 5 isolates (TgPigSp1 to TgPigSp5) were obtained. The isolates were characterized by using 11 PCR-RFLP genetic markers; three isolates had a ToxoDB #3 genotype (3/5) and two isolates had a ToxoDB #2 genotype (2/5). The TgPigSp1 and TgPigSp4 isolates were selected for virulence in mice characterization as instances of each different RFLP-genotype found. The TgPigSp1 isolate (#2 genotype) was virulent in mice with notable cumulative mortality (87.5%) and morbidity rates (100%); the TgPigSp4 (#3) was nonvirulent and triggered mild clinical signs in 42.1% of seropositive mice. Infection dynamics and organ distribution of both isolates were analyzed; the data revealed significant differences, including substantially higher parasite load in the lung during the acute phase of infection, in mice infected with TgPigSp1 than in the case of TgPigSp4 (median parasite load 7.6 vs. 0 zoites/mg, respectively; p < 0.05). Furthermore, degrees of severity of detected histopathological lesions appeared to be related to higher parasite burdens. Taking into account the unexpectedly high mortality rate and parasite load associated with the clonal genotype III, which is traditionally considered nonvirulent in mice, the need for further investigation and characterization of the T. gondii strains circulating in any host in Europe is emphasized.
| S-EPMC7714755 | biostudies-literature
Project description:Sheep are highly susceptible to Toxoplasma gondii, and miscarriage is the main clinical feature. This study investigated 227 sheep samples (210 myocardial tissues from slaughterhouses, 6 ewe serum samples, 3 aborted fetuses, and 8 dead lambs from veterinary clinics) from central China for T. gondii infection. Antibodies against T. gondii were detected using the modified agglutination test (MAT). PCR was performed to detect T. gondii DNA in the tissue samples. The results showed that four samples were seropositive (MAT titer ≥ 1:100), with a seroprevalence of 1.8% (4/227). The seropositive samples included two myocardial samples from a slaughterhouse, one ewe and its aborted fetus from a veterinary clinic. The results revealed that 7 out of 207 (3.4%) sheep tissue samples were PCR-positive, including two myocardial tissue samples from slaughterhouses, three aborted fetuses, and two lambs from veterinary clinics. Toxoplasma gondii vertical transmission had occurred in two of three pairs of ewes and her pups. One viable T. gondii strain (TgSheepCHn14) was isolated from the myocardial tissues of sheep from a slaughterhouse. Tachyzoites were obtained from cell cultures at 70 days following seeding in the brains and lungs of mice. This strain was non-lethal to Swiss mice. The number of parasite brain cysts in mice decreased with time post-infection (p < 0.05). Overall, the prevalence of T. gondii in the sheep samples was low. Although the samples were scattered, and not from planned collections, the current study detected T. gondii antibodies and DNA in aborted fetuses, indicating that vertical transmission could occur and maintain the parasites in sheep herds without exogenous infection.
| S-EPMC10302579 | biostudies-literature
Project description:Aminopeptidases have emerged as new promising drug targets for the development of novel anti-parasitic drugs. An aspartyl aminopeptidase-like gene has been identified in the Toxoplasma gondii genome (TgAAP), although its function remains unknown. In this study, we characterized TgAAP and performed functional analysis of the gene product. Firstly, we expressed a functional recombinant TgAAP (rTgAAP) protein in Escherichia coli, and found that it required metal ions for activity and showed a substrate preference for N-terminal acidic amino acids Glu and Asp. Then, we evaluated the function and drug target potential of TgAAP using the CRISPR/Cas9 knockout system. Western blotting demonstrated the deletion of TgAAP in the knockout strain. Indirect immunofluorescence analysis showed that TgAAP was localized in the cytoplasm of the wild-type parasite, but was not expressed in the knockout strain. Phenotype analysis revealed that TgAAP knockout inhibited the attachment/invasion, replication, and substrate-specific activity in T. gondii. Finally, the activity of drug CID 23724194, previously described as targeting Plasmodium and malarial parasite AAP, was tested against rTgAAP and the parasite. Overall, TgAAP knockout affected the growth of T. gondii but did not completely abolish parasite replication and growth. Therefore, TgAAP may comprise a useful adjunct drug target of T. gondii.
| S-EPMC5039622 | biostudies-literature