32 results on '"Bernal, Rafael"'
Search Results
2. In vivo and in vitro models show unexpected degrees of virulence among Toxoplasma gondii type II and III isolates from sheep
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Fernández-Escobar, Mercedes, Calero-Bernal, Rafael, Regidor-Cerrillo, Javier, Vallejo, Raquel, Benavides, Julio, Collantes-Fernández, Esther, and Ortega-Mora, Luis Miguel
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- 2021
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3. Molecular survey for cyst-forming coccidia (Toxoplasma gondii, Neospora caninum, Sarcocystis spp.) in Mediterranean periurban micromammals
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Fernández-Escobar, Mercedes, Millán, Javier, Chirife, Andrea D., Ortega-Mora, Luis Miguel, and Calero-Bernal, Rafael
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- 2020
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4. Anti- Toxoplasma gondii Antibodies in European Residents: A Systematic Review and Meta-Analysis of Studies Published between 2000 and 2020.
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Calero-Bernal, Rafael, Gennari, Solange María, Cano, Santiago, Salas-Fajardo, Martha Ynés, Ríos, Arantxa, Álvarez-García, Gema, and Ortega-Mora, Luis Miguel
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TOXOPLASMA gondii ,BEEF industry ,PUBLICATION bias ,SEROPREVALENCE ,ANIMAL health - Abstract
Toxoplasmosis has a major impact on animal and public health. Information regarding the seroprevalence of human Toxoplasma gondii infections from a European perspective has not yet been compiled to date. Thus, the present review summarized available resident data from the period 2000–2020. The overall seroprevalence of anti-T. gondii IgG was 32.1%, with great variability between countries (n = 30). The subgroup analysis identified different pooled prevalence data depending on the geographic area (p < 0.0001), target population (p = 0.0147), and serological diagnosis assays used (p = 0.0059). A high heterogeneity (I
2 = 100%, p < 0.001; Q = 3.5e+05, d.f. = 135, p < 0.001) and degree of publication bias (Egger's test = 6.14, p < 0.001) were observed among the 134 studies considered. The occurrence of anti-T. gondii IgM, which was reported in 64.7% of studies, reached a pooled seroprevalence of 0.6%. In addition, among the eight main risk factors identified, "contact with soil", "consumption of undercooked beef", and "intake of unwashed vegetables" were the most significantly associated with infections. The fact that one-third of the European population has been exposed to T. gondii justifies extra efforts to harmonize surveillance systems and develop additional risk-factor analyses based on detailed source attribution assessment. [ABSTRACT FROM AUTHOR]- Published
- 2023
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5. Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks
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Fernández-Escobar, Mercedes, Calero-Bernal, Rafael, Benavides, Julio, Regidor-Cerrillo, Javier, Guerrero-Molina, María Cristina, Gutiérrez-Expósito, Daniel, Collantes-Fernández, Esther, and Ortega-Mora, Luis Miguel
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- 2020
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6. D-JRP-TOXOSOURCES-WP3.5 Report on occurrence of T. gondii in selected fresh produce in different regions in Europe
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Lalle, Marco, Betson, Martha, Alvarez Garcia, Gema, Calero Bernal, Rafael, and Jokelainen, Pikka
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Toxoplasma gondii ,TOXOSOURCES - Abstract
DeliverableD-JRP-TOXOSOURCES-WP3.5 Report on occurrence of T. gondii in selected fresh produce in different regions in Europe
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- 2022
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7. Deliverable D-JRP-TOXOSOURCES-WP5.3 Report on the inter-laboratory comparison and the pilot studies of WP5
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Joeres, Maike, Calero Bernal, Rafael, Maksimov, Pavlo, Caccio, Simone, Ortega-Mora, Luis, Jokelainen, Pikka, Schares, Gereon, and TOXOSOURCES Consortium
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Toxoplasma gondii ,TOXOSOURCES - Abstract
Deliverable D-JRP-TOXOSOURCES-WP5.3 Report on the inter-laboratory comparison and the pilot studies of WP5
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- 2022
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8. Seroprevalence and genetic characterization of Toxoplasma gondii in domestic pigs intended for human consumption in Cuba.
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Castillo‐Cuenca, Julio César, Almería, Sonia, Calero‐Bernal, Rafael, Fernández‐Escobar, Mercedes, Fraga, Jorge, Entrena‐García, Angel, Arias, Pedro Casanova, Martínez‐Moreno, Álvaro, and García‐Bocanegra, Ignacio
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SWINE ,TOXOPLASMA gondii ,SWINE farms ,SEROPREVALENCE ,GENETIC profile ,IMMUNOGLOBULINS ,SWINE breeding ,PORK products - Abstract
Domestic pigs are considered as one of the main intermediate hosts in the zoonotic transmission of Toxoplasma gondii in many countries. Serological and molecular studies are warranted to better understand the epidemiology and transmission patterns of this parasite worldwide. To date, seroepidemiological information on T. gondii in domestic pigs in Cuba is very scarce and there are no reports of T. gondii genotypes circulating in this country. Here, we aimed to estimate the seroprevalence of T. gondii and provide genetic characterization of the strains circulating in slaughtered pigs intended for human consumption in Central Cuba. Seroprevalence was determined in 450 serum samples from slaughtered pigs in Villa Clara province using ELISA. Anti‐Toxoplasma gondii IgG antibodies were detected in 100 animals (22.2%, 95% CI: 18.5–26.2). Conventional PCR of the 529‐bp marker of T. gondii was performed in hearts and diaphragm tissues of all ELISA‐seropositive pigs. Toxoplasma gondii DNA was detected in four animals. Further genetic characterization of the positive DNA samples was performed by multilocus PCR‐RFLP and PCR‐sequencing typing tools. Molecular analysis revealed four different genetic profiles that were combinations of type I, II, III and u‐1 alleles, suggesting the circulation of non‐clonal genotypes of T. gondii in domestic pigs in Cuba. Our results indicate that T. gondii is widely distributed in slaughtered pigs in this country, which might have important implications for public health. To the best of our knowledge, this is the first report on genetic characterization of T. gondii in Cuba. Although preliminary, the results suggest a high genetic diversity of T. gondii in the study region. Further studies based on parasite isolation are needed to definitively identify the genotypes circulating and characterize the virulence of strains detected in pigs in Cuba, and to assess the risk of zoonotic transmission from pork products in this country. [ABSTRACT FROM AUTHOR]
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- 2023
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9. Toxoplasmosis in geese and detection of two new atypical Toxoplasma gondii strains from naturally infected Canada geese (Branta canadensis)
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Verma, Shiv Kumar, Calero-Bernal, Rafael, Cerqueira-Cézar, Camila K., Kwok, Oliver C. H., Dudley, Mike, Jiang, Tiantian, Su, Chunlei, Hill, Dolores, and Dubey, Jitender P.
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- 2016
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10. Seroprevalence, isolation, first genetic characterization of Toxoplasma gondii, and possible congenital transmission in wild moose from Minnesota, USA
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Verma, Shiv K., Carstensen, Michelle, Calero-Bernal, Rafael, Moore, Seth A., Jiang, Tiantian, Su, Chunlei, and Dubey, Jitender P.
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- 2016
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11. Deliverable D-JRP-TOXOSOURCES-WP2.2 Report on quantitative exposure data from survey of WP2
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Opsteegh, Marieke, Swart, Arno, Chardon, Jurgen, Pires, Sara M., Bier, Nadja, Mayer-Scholl, Anne, Schares, Gereon, Jore, Solveig, Davidson, Rebecca, Waap, Helga, Calero Bernal, Rafael, Alvarez Garcia, Gema, Blaga, Radu, Damek, Filip, LeRoux, Delphine, Stensvold, Christen R., Sroka, Jacek, Rozycki, Miros��aw, Koudela, B��etislav, Ottoson, Jakob, Lalle, Marco, de Haas, Mattijs, Beninc��, Elisa, van der Giessen, Joke, and Jokelainen, Pikka
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Toxoplasma gondii ,TOXOSOURCES - Abstract
Deliverable D-JRP-TOXOSOURCES-WP2.2 Report on quantitative exposure data from survey of WP2
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- 2021
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12. Deliverable D-JRP-TOXOSOURCES-WP5.1 SOP for a novel NGS-MLST genotyping method of WP5
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Schares, Gereon, Maksimov, Pavlo, Joeres, Maike, Caccio, Simone, Vatta, Paolo, ��lvarez Garc��a, Gema, Ortega Mora, Luis M., Calero-Bernal, Rafael, Fernandez Escobar, Mercedes, Angulo Lara, Pablo, Blaga, Radu, Koudela, B��etislav, and Jokelainen, Pikka
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Toxoplasma gondii ,TOXOSOURCES - Abstract
Deliverable D-JRP-TOXOSOURCES-WP5.1 SOP for a novel NGS-MLST genotyping method of WP5
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- 2021
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13. Archetypal type II and III Toxoplasma gondii oocysts induce different immune responses and clinical outcomes in experimentally infected piglets.
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Largo-de la Torre, Andrea, Diezma-Díaz, Carlos, Calero-Bernal, Rafael, Atencia-Cibreiro, Gabriela, Sánchez-Sánchez, Roberto, Ferre, Ignacio, Regidor-Cerrillo, Javier, and Miguel Ortega-Mora, Luis
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OOCYSTS ,TOXOPLASMA gondii ,PIGLETS ,IMMUNE response ,TREATMENT effectiveness ,SYMPTOMS - Abstract
Livestock animals, such as swine, are an important source of Toxoplasma gondii in the human population. Currently, there is limited knowledge regarding the potential influence that the T. gondii genotype might exert on establishing infection in swine. Herein, we investigated the role of 2 T. gondii isolates, type II and III, representative of the genotypes circulating in Europe, in the immune responses and infection dynamics in piglets. Recently obtained oocysts (103) from the T. gondii field isolates TgShSp1 (type II, ToxoDB genotype #3) and TgShSp24 (type III, #2) were used for oral infection. Thirteen 50-day-old female piglets of the Landrace-Large White crossbreed were randomly allocated into three different groups: Group 1 (G1, n=5), inoculated with TgShSp1; Group 2 (G2, n=5), inoculated with TgShSp24; and Group 3 (G3, n=3), a non-infected control group. Clinical signs were monitored daily until 42 days post-infection (dpi) when piglets were euthanized. Blood samples were collected weekly to test the cellular immune response in parasite-stimulated peripheral blood and specific IgG, IgG1 and IgG2, responses in sera. Parasite distribution and burden were evaluated in target tissues using a mouse bioassay and quantitative RT-PCR (qPCR). Apathy and a moderate decrease in feed consumption were observed in G1 and G2 piglets between 5 and 8 dpi, coinciding with fever (>40°C). G2 piglets had higher temperatures for a longer duration. Using mouse bioassay and qPCR, the detection frequency was higher in G2 vs. G1, and the highest parasite burdens in target tissues were also found in G2. Seroconversion was detected at 14 dpi in both infected groups, but higher antibody levels were observed in G2 piglets. Cytokine analyses revealed the production of IL-8, IL-1β and IFN-ɤ from 7 dpi in both infected groups. Moreover, IL-12 was produced from 7 dpi in G1 and from 14 dpi in G2. Levels of IL-8 were higher in G2, but IL-1β, IL-12 and IFN-ɤ were higher in G1 at 14 dpi. This cytokine profile reveals a predominant proinflammatory response that could be involved in limiting T. gondii infection in piglets, although it is more efficient against TgShSp1 type II-driven infection. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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14. Toxoplasma gondii Genetic Diversity in Mediterranean Dolphins.
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Fernández-Escobar, Mercedes, Giorda, Federica, Mattioda, Virgina, Audino, Tania, Di Nocera, Fabio, Lucifora, Giuseppe, Varello, Katia, Grattarola, Carla, Ortega-Mora, Luis Miguel, Casalone, Cristina, and Calero-Bernal, Rafael
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GENETIC variation ,TOXOPLASMA gondii ,BOTTLENOSE dolphin ,DOLPHINS ,MIXED infections ,NEUROLOGICAL disorders ,CETACEA - Abstract
Toxoplasma gondii constitutes a major zoonotic agent but also has been frequently identified as an important cause of clinical disease (e.g., abortion, pneumonia, encephalitis) in wildlife; specifically, T. gondii has been associated with neurological disease in cetaceans. This study investigated the genetic diversity of T. gondii strains involved in infections in dolphins found stranded in the Mediterranean coastlines of Italy. Tissue samples from 16 dolphins (Stenella coeruleoalba and Tursiops truncatus species) positive for T. gondii-DNA presence by PCR were examined by histology and subjected to further genetic characterization of strains detected by PCR-RFLP and multilocus PCR-sequencing assays. According to fully genotyped samples, the genotypes ToxoDB#3 (67%) and #2 (22%) were detected, the latter being reported for the first time in cetaceans, along with a mixed infection (11%). Subtyping by PCR-seq procedures provided evidence of common point mutations in strains from southwestern Europe. Despite evidence of T. gondii as a cause of neurological disease in dolphins, sources of infections are difficult to identify since they are long-living animals and some species have vast migration areas with multiple chances of infection. Finally, the genetic diversity of T. gondii found in the dolphins studied in the Mediterranean coastlines of Italy reflects the main genotypes circulating inland in the European continent. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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15. Unifying Virulence Evaluation in Toxoplasma gondii : A Timely Task.
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Calero-Bernal, Rafael, Fernández-Escobar, Mercedes, Katzer, Frank, Su, Chunlei, and Ortega-Mora, Luis Miguel
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TOXOPLASMA gondii ,LABORATORY mice ,GENETIC variation ,DEATH rate ,PHENOTYPES ,TREATMENT effectiveness - Abstract
Toxoplasma gondii , a major zoonotic pathogen, possess a significant genetic and phenotypic diversity that have been proposed to be responsible for the variation in clinical outcomes, mainly related to reproductive failure and ocular and neurological signs. Different T. gondii haplogroups showed strong phenotypic differences in laboratory mouse infections, which provide a suitable model for mimicking acute and chronic infections. In addition, it has been observed that degrees of virulence might be related to the physiological status of the host and its genetic background. Currently, mortality rate (lethality) in outbred laboratory mice is the most significant phenotypic marker, which has been well defined for the three archetypal clonal types (I, II and III) of T. gondii ; nevertheless, such a trait seems to be insufficient to discriminate between different degrees of virulence of field isolates. Many other non-lethal parameters, observed both in in vivo and in vitro experimental models, have been suggested as highly informative, yielding promising discriminatory power. Although intra-genotype variations have been observed in phenotypic characteristics, there is no clear picture of the phenotypes circulating worldwide; therefore, a global overview of T. gondii strain mortality in mice is presented here. Molecular characterization has been normalized to some extent, but this is not the case for the phenotypic characterization and definition of virulence. The present paper proposes a baseline (minimum required information) for the phenotypic characterization of T. gondii virulence and intends to highlight the needs for consistent methods when a panel of T. gondii isolates is evaluated for virulence. [ABSTRACT FROM AUTHOR]
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- 2022
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16. Toxoplasma gondii Genotyping: A Closer Look Into Europe.
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Fernández-Escobar, Mercedes, Schares, Gereon, Maksimov, Pavlo, Joeres, Maike, Ortega-Mora, Luis Miguel, and Calero-Bernal, Rafael
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TOXOPLASMA gondii ,IMMUNOCOMPROMISED patients ,GENOTYPES - Abstract
Toxoplasma gondii is a major zoonotic agent which may cause harmful effects mainly in pregnant and immunocompromised hosts. Despite many efforts on its genetic characterization, an entirely clear picture of the population structure in Europe has not been achieved yet. The present study aimed to summarize the available genotyping information and to map the distribution of circulating strains. There is consensus on type II T. gondii genotypes prevailing in Europe, but the absence of harmonization in the use of typing methods limits detailed knowledge. Standardized, high-end typing tools and integrative strategies are needed to fill the gaps and complete an accurate image of the T. gondii genetic population in Europe. [ABSTRACT FROM AUTHOR]
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- 2022
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17. Contamination of Soil, Water, Fresh Produce, and Bivalve Mollusks with Toxoplasma gondii Oocysts: A Systematic Review.
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López Ureña, Nadia María, Chaudhry, Umer, Calero Bernal, Rafael, Cano Alsua, Santiago, Messina, Davide, Evangelista, Francisco, Betson, Martha, Lalle, Marco, Jokelainen, Pikka, Ortega Mora, Luis Miguel, and Álvarez García, Gema
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SOIL pollution ,TOXOPLASMA gondii ,MOLLUSKS ,OOCYSTS ,WARM-blooded animals - Abstract
Toxoplasma gondii is a major foodborne pathogen capable of infecting all warm-blooded animals, including humans. Although oocyst-associated toxoplasmosis outbreaks have been documented, the relevance of the environmental transmission route remains poorly investigated. Thus, we carried out an extensive systematic review on T. gondii oocyst contamination of soil, water, fresh produce, and mollusk bivalves, following the PRISMA guidelines. Studies published up to the end of 2020 were searched for in public databases and screened. The reference sections of the selected articles were examined to identify additional studies. A total of 102 out of 3201 articles were selected: 34 articles focused on soil, 40 focused on water, 23 focused on fresh produce (vegetables/fruits), and 21 focused on bivalve mollusks. Toxoplasma gondii oocysts were found in all matrices worldwide, with detection rates ranging from 0.09% (1/1109) to 100% (8/8) using bioassay or PCR-based detection methods. There was a high heterogeneity (I
2 = 98.9%), which was influenced by both the sampling strategy (e.g., sampling site and sample type, sample composition, sample origin, season, number of samples, cat presence) and methodology (recovery and detection methods). Harmonized approaches are needed for the detection of T. gondii in different environmental matrices in order to obtain robust and comparable results. [ABSTRACT FROM AUTHOR]- Published
- 2022
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18. Toxoplasma gondii infection in domestic and wild felids as public health concerns: a systematic review and meta-analysis.
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Hatam-Nahavandi, Kareem, Calero-Bernal, Rafael, Rahimi, Mohammad Taghi, Pagheh, Abdol Sattar, Zarean, Mehdi, Dezhkam, Asiyeh, and Ahmadpour, Ehsan
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TOXOPLASMA gondii , *FELIDAE , *OOCYSTS , *SEROPREVALENCE , *SYSTEMATIC reviews - Abstract
Felidae as definitive hosts for Toxoplasma gondii play a major role in transmission to all warm-blooded animals trough oocysts dissemination. Therefore the current comprehensive study was performed to determine the global status of T. gondii infection in domestic and wild felids aiming to provide comprehensive data of interest for further intervention approaching the One Health perspective. Different databases were searched by utilizing particular key words for publications related to T. gondii infecting domestic and wild feline host species, worldwide, from 1970 to 2020. The review of 337 reports showed that the seroprevalence of T. gondii in domestic cats and wild felids was estimated in 37.5% (95% CI 34.7–40.3) (I2 = 98.3%, P < 0.001) and 64% (95% CI 60–67.9) (I2 = 88%, P < 0.0001), respectively. The global pooled prevalence of oocysts in the fecal examined specimens from domestic cats was estimated in 2.6% (95% CI 1.9–3.3) (I2 = 96.1%, P < 0.0001), and that in fecal samples from wild felids was estimated in 2.4% (95% CI 1.1–4.2) (I2 = 86.4%, P < 0.0001). In addition, from 13,252 examined soil samples in 14 reviewed studies, the pooled occurrence of T. gondii oocysts was determined in 16.2% (95% CI 7.66–27.03%). The observed high rates of anti-T. gondii antibodies seroprevalence levels and oocyst excretion frequency in the felids, along with soil (environmental) contamination with oocysts may constitute a potential threat to animal and public health, and data will result of interest in further prophylaxis programs. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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19. Optimization of the most widely used serological tests for a harmonized diagnosis of Toxoplasma gondii infection in domestic pigs.
- Author
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López-Ureña, Nadia María, Calero-Bernal, Rafael, González-Fernández, Nuria, Blaga, Radu, Koudela, Bretislav, Ortega-Mora, Luis Miguel, and Álvarez-García, Gema
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SWINE , *SERODIAGNOSIS , *TOXOPLASMA gondii , *IMMUNOGLOBULINS , *ENZYME-linked immunosorbent assay , *ANTIBODY titer - Abstract
The intake of Toxoplasma gondii tissue cysts through raw or undercooked pork meat is one of the main infection sources for humans. Thus, surveillance is recommended to control and prevent infection in domestic pigs. However, the lack of comparative studies hampers the updating of their performance and the comparison of seroprevalence data. Therefore, the aim of this study was to develop and validate three in-house tests and accomplish a comparative analysis of the most widely used serological tests employed in pigs. A panel of sera from pigs experimentally infected with either oocysts or tissue cysts from type II and III isolates (n = 158) was used to develop and validate a tachyzoite-based Western blot assay. Then, this technique was used as a reference to develop and preliminary validate a lyophilized tachyzoite-based enzyme-linked immunosorbent assay and an immunofluorescence antibody test. Next, a comparative study of the three in-house tests and three widely used commercial ELISAs (IDScreen®, PrioCHECK™ and Pigtype®) was accomplished with the abovementioned sera together with an additional serum panel of pigs experimentally infected with oocysts from the type II isolate (n = 44) and a panel of naturally infected pigs (n = 244). The results obtained by the majority of the tests were regarded as reference, and data analyses included TG-ROC calculations and agreement tests. Finally, the kinetics of anti- T. gondii IgGs from experimentally infected pigs was analyzed. Excellent sensitivity (Se) and specificity (Sp) values (≥ 93%) and moderate to near perfect agreement (k = 0.63–0.91) were observed using sera from experimental infections without requiring further readjustment, except for PrioCHECK (100% Se, 73% Sp). However, the Se of IDScreen® (87%) and TgSALUVET WB (71%) and the Sp of PrioCHECK (72%) were slightly or notably reduced when sera from naturally infected animals were analyzed, which also influenced the kappa values (k = 0.30–0.91). Cutoff readjustments increased the Se and Sp values to equal to or above 97% for all tests, except for TgSALUVET WB, which can be used as a reference for initial validation of tests, but it is not recommended for routine diagnosis. Seroconversion was recorded from two weeks post-infection by most of the tests, with significantly higher IgG levels in sera from pigs infected with the T. gondii type III vs. type II isolate. Again, differences regarding the test employed were observed. Differences in the diagnostic performance among tests evidenced the need to harmonize serological techniques to obtain comparable and reliable results. • Western blot is an appropriate reference test for initial technique validation. • A comparative study of commercial and in-house tests led to harmonized serological results. • Pigtype ELISA, TgSALUVET ELISA 2.0 and TgSALUVET IFAT showed excellent diagnostic performance. • The PrioCHECK and IDScreen ELISA tests required further readjustments to increase Se and Sp. • Higher IgG levels were detected in pigs infected with type III vs. type II isolates by most of the tests. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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20. Contribución de los métodos serológicos y moleculares al conocimiento de la epidemiología de 'Toxoplasma gondii' en Extremadura
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Calero Bernal, Rafael, Frontera Carrión, Eva María, Pérez Martín, Juan Enrique, Reina Esojo, David, and Universidad de Extremadura. Departamento de Sanidad Animal
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Extremadura ,Epidemiology ,Toxoplasma gondii ,Epidemiología ,Parasitología ,Parasitology - Abstract
Toxoplasma gondii es uno de los parásitos más difundidos en la naturaleza, por lo que requiere grandes esfuerzos para su prevención y control. La finalidad de este estudio es la aplicación de métodos serológicos (ELISA) y moleculares (PCR anidada, PCR a tiempo real y RFLP-PCR) para conocer la epidemiología de T. gondii en los ecosistemas del Suroeste español y determinar la prevalencia del parásito y los genotipos presentes en los diferentes hospedadores contemplados. En relación a la población humana, el estudio ha pretendido conocer la situación de la enfermedad en Extremadura, y las caracterizar muestras provenientes de casos clínicos, determinando los tipos genéticos circulantes en España. Tras realizar los correspondientes análisis serológicos en 8 especies hospedadoras, moleculares de detección en 10 y de caracterización en 9, incluyendo la humana. El estudio reveló una gran presencia de T. gondii en las comarcas de La Siberia, La Serena, Valle del Jerte y Campo Arañuelo, pues alcanzaron las mayores prevalencias y la coexistencia de la infección, tanto en especies domésticas como silvestres, integrantes de los ciclos sinantrópicos y parantrópicos de la enfermedad. La prevalencia más alta de IgG sérica en la población humana de Extremadura se observó en el Área de Salud de Don Benito-Villanueva (20,08%), mientras que la más baja (17,56%) en el de Badajoz. Las infecciones recientes (IgM) se sitúan en niveles medios del 3,22%, siendo éstos más elevados que los encontrados en otras áreas de España. A partir de los resultados obtenidos en el presente estudio, se propone el incremento del muestreo en especies como el gamo, el zorro y los lagomorfos, la continuación en la aplicación de métodos de caracterización para obtener conclusiones más definitivas y adicionalmente, el ¿Programa Integral de Acciones de Lucha¿ propuesto debiera incluir mejoras en la declaración y el estudio de casos positivos, el incremento del cribado serológico en la población humana, el fomento de la educación para la salud y el desarrollo de estudios de coste-beneficio.
- Published
- 2013
21. Isolation and RFLP Genotyping of Toxoplasma gondii in Free-Range Chickens ( Gallus domesticus) in Grenada, West Indies, Revealed Widespread and Dominance of Clonal Type III Parasites.
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Chikweto, Alfred, Sharma, Ravindra N., Tiwari, Keshaw P., Verma, Shiv K., Calero-Bernal, Rafael, Jiang, Tiantian, Su, Chunlei, Kwok, Oliver C., and Dubey, Jitender P.
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TOXOPLASMA gondii ,RESTRICTION fragment length polymorphisms ,CHICKENS ,POULTRY parasites - Abstract
The objectives of the present cross-sectional study were to isolate and genotype Toxoplasma gondii in free-range chickens from Grenada, West Indies. Using the modified agglutination test, antibodies to T. gondii were found in 39 (26.9%) of 145 free-range chickens with titers of 25 in 7 chickens, 50 in 6 chickens, 100 in 2 chickens, and 200 or higher in 24 chickens. The hearts of the 39 seropositive chickens were bioassayed in mice; viable T. gondii was isolated from 20 and further propagated in cell culture. Genotyping of T. gondii DNA extracted from cell-cultured tachyzoites using the 10 PCR-restriction fragment length polymorphism (RFLP) markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico revealed 4 genotypes, including ToxoDB PCR-RFLP no. 2 (Type III), no. 7, no. 13, and no. 259 (new). These results indicated that T. gondii population genetics in free-range chickens seems to be moderately diverse with ToxoDB no. 2 (Type III) as the most frequent (15/20 = 75%) compared to other genotypes in Grenada. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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22. Antibody Detection and Molecular Characterization of Toxoplasma gondii from Bobcats ( Lynx rufus), Domestic Cats ( Felis catus), and Wildlife from Minnesota, USA.
- Author
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Verma, Shiv K., Minicucci, Larissa, Murphy, Darby, Carstensen, Michelle, Humpal, Carolin, Wolf, Paul, Calero‐Bernal, Rafael, Cerqueira‐Cézar, Camila K., Kwok, Oliver C.H., Su, Chunlei, Hill, Dolores, and Dubey, Jitender P.
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TOXOPLASMA gondii ,BOBCAT ,ANIMALS ,CELL culture ,AGGLUTINATION tests - Abstract
Little is known of the epidemiology of toxoplasmosis in Minnesota. Here, we evaluated Toxoplasma gondii infection in 50 wild bobcats ( Lynx rufus) and 75 other animals on/near 10 cattle farms. Antibodies to T. gondii were assayed in serum samples or tissue fluids by the modified agglutination test (MAT, cut-off 1:25). Twenty nine of 50 bobcats and 15 of 41 wildlife trapped on the vicinity of 10 farms and nine of 16 adult domestic cats ( Felis catus) and six of 14 domestic dogs resident on farms were seropositive. Toxoplasma gondii oocysts were not found in feces of any felid. Tissues of all seropositive wild animals trapped on the farm were bioassayed in mice and viable T. gondii was isolated from two badgers ( Taxidea taxus), two raccoons ( Procyon lotor), one coyote ( Canis latrans), and one opossum ( Didelphis virginiana). All six T. gondii isolates were further propagated in cell culture. Multi-locus PCR-RFLP genotyping using 10 markers (SAG1, SAG2 (5′-3′SAG2, and alt.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico), and DNA from cell culture derived tachyzoites revealed three genotypes; #5 ToxoDataBase (1 coyote, 1 raccoon), #1 (1 badger, 1 raccoon, 1 opossum), and #2 (1 badger). This is the first report of T. gondii prevalence in domestic cats and in bobcats from Minnesota, and the first isolation of viable T. gondii from badger. [ABSTRACT FROM AUTHOR]
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- 2016
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23. Short-term culture adaptation of Toxoplasma gondii archetypal II and III field isolates affects cystogenic capabilities and modifies virulence in mice.
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Colos-Arango, Alicia, Largo-de la Torre, Andrea, Calero-Bernal, Rafael, Ortega-Mora, Luis-Miguel, and Regidor-Cerrillo, Javier
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- *
TOXOPLASMA gondii , *ARACHNOID cysts , *LYTIC cycle , *CELL culture , *CULTURAL maintenance , *MICE , *INFECTION control - Abstract
[Display omitted] • The influence of in vitro maintenance on phenotypic traits of Toxoplasma gondii was studied. • Cell culture adaptation in the short term influences cyst formation and mouse virulence. • Toxoplasma gondii isolates experience a loss of cyst formation after 40 culture passages. • Changes in mouse virulence correlated with in vitro parasite growth and cyst formation. Most Toxoplasma gondii research has been carried out using strains maintained in the laboratory for long periods of time. Long-term passage in mice or cell culture influences T. gondii phenotypic traits such as the capability to produce oocysts in cats and virulence in mice. In this work, we investigated the effect of cell culture adaptation in the short term for recently obtained type II (Tg ShSp1 (Genotype ToxoDB#3), Tg ShSp2 (#1), Tg ShSp3 (#3) and Tg ShSp16 (#3)) and type III (#2) isolates (Tg ShSp24 and Tg PigSp1). With this purpose, spontaneous and alkaline stress-induced cyst formation in Vero cells during 40 passages, from passage 10 (p 10) to 50 (p 50), and isolate virulence at p 10 versus p 50 were studied using a harmonized bioassay method in Swiss/CD1 mice. T. gondii cell culture maintenance showed a drastic loss of spontaneous and induced production of mature cysts after ≈25–30 passages. The Tg ShSp1, Tg ShSp16 and Tg ShSp24 isolates failed to generate spontaneously formed mature cysts at p 50. Limited cyst formation was associated with an increase in parasite growth and a shorter lytic cycle. In vitro maintenance also modified T. gondii virulence in mice at p 50 with events of exacerbation, increasing cumulative morbidity for Tg ShSp2 and Tg ShSp3 isolates and mortality for Tg ShSp24 and Tg PigSp1 isolates, or attenuation, with absence of mortality and severe clinical signs for Tg ShSp16, and better control of the infection with the lowest parasite and cyst burdens in lungs and brain for the Tg ShSp1 isolate. The present findings show deep changes in relevant phenotypic traits in laboratory-adapted T. gondii isolates and open new discussion about their use for inferring keys to parasite biology and virulence. [ABSTRACT FROM AUTHOR]
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- 2023
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24. Isolation and Genetic Characterization of Toxoplasma gondii from Black Bears ( Ursus americanus), Bobcats ( Lynx rufus), and Feral Cats ( Felis catus) from Pennsylvania.
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Dubey, Jitender P., Verma, Shiv K., Calero‐Bernal, Rafael, Cassinelli, Ana B., Kwok, Oliver C. H., Van Why, Kyle, Su, Chunlei, and Humphreys, Jan G.
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TOXOPLASMA gondii ,BLACK bear ,BOBCAT ,FERAL cats ,WARM-blooded animals - Abstract
Toxoplasma gondii infects virtually all warm-blooded hosts worldwide. Recently, attention has been focused on the genetic diversity of the parasite to explain its pathogenicity in different hosts. It has been hypothesized that interaction between feral and domestic cycles of T. gondii may increase unusual genotypes in domestic cats and facilitate transmission of potentially more pathogenic genotypes to humans, domestic animals, and wildlife. In the present study, we tested black bear ( Ursus americanus), bobcat ( Lynx rufus), and feral cat ( Felis catus) from the state of Pennsylvania for T. gondii infection. Antibodies to T. gondii were found in 32 (84.2%) of 38 bears, both bobcats, and 2 of 3 feral cats tested by the modified agglutination test (cut off titer 1:25). Hearts from seropositive animals were bioassayed in mice, and viable T. gondii was isolated from 3 of 32 bears, 2 of 2 bobcats, and 2 of 3 feral cats. DNA isolated from culture-derived tachyzoites of these isolates was characterized using multilocus PCR- RFLP markers. Three genotypes were revealed, including Toxo DB PCR- RFLP genotype #1 or #3 (Type II, 1 isolate), #5 (Type 12, 3 isolates), and #216 (3 isolates), adding to the evidence of genetic diversity of T. gondii in wildlife in Pennsylvania. Pathogenicity of 3 T. gondii isolates (all #216, 1 from bear, and 2 from feral cat) was determined in outbred Swiss Webster mice; all three were virulent causing 100% mortality. Results indicated that highly mouse pathogenic strains of T. gondii are circulating in wildlife, and these strains may pose risk to infect human through consuming of game meat. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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25. Prevalence and Genotype Identification of Toxoplasma gondii in Wild Animals from Southwestern Spain.
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Calero-Bernal, Rafael, Saugar, José M., Frontera, Eva, Pérez-Martín, Juan E., Habela, Miguel A., Serrano, Francisco J., Reina, David, and Fuentes, Isabel
- Abstract
The article reports on the prevalence of Toxoplasma gondii among wild animals in the southwestern area of Spain. Topics discussed include the use of polymerase chain reaction (PCR) in detecting the zoonotic pathogen in wild animals, the animals that tested positive for T. gondii and the implication of the genetic variability produced by the test results for domestic animal and human infection.
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- 2015
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26. Congenital Toxoplasmosis in Wild Boar (Sus scrofa) and Identification of the Toxoplasma gondii Types Involved.
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Calero-Bernal, Rafael, Gómez-Gordo, Luis, Saugar, José María, Frontera, Eva, Pérez-Martin, Juan Enrique, Reina, David, Serrano, Francisco Javier, and Fuentes, Isabel
- Abstract
The article discusses a study that reported a case of vertical transmission of congenital toxoplasmosis in wild boar (Sus scrofa). Lesions compatible with acute toxoplasmosis were identified in a pregnant female and her three fetuses following necropsy and histopathologic examination. Toxoplasma gondii in maternal and fetal samples were detected. Findings suggest that the pathogenicity of the various T. gondii types may differ in wild boars.
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- 2013
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27. Evidence for Unknown Sarcocystis -Like Infection in Stranded Striped Dolphins (Stenella coeruleoalba) from the Ligurian Sea, Italy.
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Giorda, Federica, Romani-Cremaschi, Umberto, Marsh, Antoinette E., Grattarola, Carla, Iulini, Barbara, Pautasso, Alessandra, Varello, Katia, Berio, Enrica, Gazzuola, Paola, Marsili, Letizia, Di Francesco, Cristina E., Goria, Maria, Verna, Federica, Audino, Tania, Peletto, Simone, Caramelli, Maria, Fernández-Escobar, Mercedes, Sierra, Eva, Fernández, Antonio, and Calero-Bernal, Rafael
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STRIPED dolphin ,CETACEA ,BOTTLENOSE dolphin ,SARCOCYSTIS ,NEOSPORA caninum ,MAMMAL conservation ,MARINE mammals ,MYOCARDIUM - Abstract
Simple Summary: Two stranded striped dolphins presented meningoenchepalitic lesions associated with the presence of unknown protozoan tissue cysts. The present study aimed at fully characterizing these previously undescribed parasites. Light microscopy re-examination of affected CNS areas showed high numbers of tissue cysts with morphological features resembling those of Sarcocystis species. Tissue cyst bradyzoites positively stained when labeled with polyclonal antisera but cross-reactivity could not be precluded. Sarcocystis sp. sequences with high homology to species infecting livestock were amplified by means of PCR from myocardial and muscle tissues. This is the first report of Sarcocystis-like tissue cysts in the cerebral tissue of stranded cetaceans with muscular sarcocystosis in Mediterranean dolphins. The obtained results may suggest a land-to-sea cycling of Apicomplexan parasites in this region and the need for further investigations in order to foster marine mammal conservation. Two striped dolphins (SD1, SD2), stranded along the Ligurian coast of Italy, were diagnosed with a nonsuppurative meningoencephalitis associated with previously undescribed protozoan tissue cysts. As tissue cysts were morphologically different from those of Toxoplasma gondii, additional histopathological, immunohistochemical, ultrastructural, and biomolecular investigations were performed, aiming to fully characterize the organism. Histopathology revealed the presence of large Sarcocystis-like tissue cysts, associated with limited inflammatory lesions in all CNS areas studied. IHC was inconclusive, as positive staining with polyclonal antisera did not preclude cross-reaction with other Sarcocystidae coccidia. Applied to each animal, 11 different PCR protocols precluded a neural infection by Sarcocystis neurona, Sarcocystis falcatula, Hammondia hammondi, and Neospora caninum. T. gondii coinfection was confirmed only in dolphin SD2. Sarcocystis sp. sequences, showing the highest homology to species infecting the Bovidae family, were amplified from SD1 myocardium and SD2 skeletal muscle. The present study represents the first report of Sarcocystis-like tissue cysts in the brain of stranded cetaceans along with the first description of Sarcocystis sp. infection in muscle tissue of dolphins from the Mediterranean basin. [ABSTRACT FROM AUTHOR]
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- 2021
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28. Correction to: Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks.
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Fernández-Escobar, Mercedes, Calero-Bernal, Rafael, Benavides, Julio, Regidor-Cerrillo, Javier, Guerrero-Molina, María Cristina, Gutiérrez-Expósito, Daniel, Collantes-Fernández, Esther, and Ortega-Mora, Luis Miguel
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- *
TOXOPLASMA gondii , *SHEEP - Abstract
An amendment to this paper has been published and can be accessed via the original article. [ABSTRACT FROM AUTHOR]
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- 2020
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29. Deliverable D-JRP-TOXOSOURCES-WP3.4 Report on literature review on the prevalence of T. gondii oocysts in fresh produce and environment
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López Ureña, Nadia María, Álvarez García, Gema, Calero Bernal, Rafael, Betson, Martha, Lalle, Marco, and Jokelainen, Pikka
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Toxoplasma gondii ,TOXOSOURCES ,3. Good health - Abstract
Deliverable D-JRP-TOXOSOURCES-WP3.4 Report on literature review on the prevalence of T. gondii oocysts in fresh produce and environment
30. Deliverable D-JRP-TOXOSOURCES-WP3.4 Report on literature review on the prevalence of T. gondii oocysts in fresh produce and environment
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L��pez Ure��a, Nadia Mar��a, ��lvarez Garc��a, Gema, Calero Bernal, Rafael, Betson, Martha, Lalle, Marco, and Jokelainen, Pikka
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Toxoplasma gondii ,TOXOSOURCES ,3. Good health - Abstract
Deliverable D-JRP-TOXOSOURCES-WP3.4 Report on literature review on the prevalence of T. gondii oocysts in fresh produce and environment
31. Experimental infection of sheep at mid-pregnancy with archetypal type II and type III Toxoplasma gondii isolates exhibited different phenotypic traits.
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Vallejo, Raquel, Benavides, Julio, Arteche-Villasol, Noive, Sánchez-Sánchez, Roberto, Calero-Bernal, Rafael, Ferreras, Ma Carmen, Criado, Miguel, Pérez, Valentín, Ortega-Mora, Luis Miguel, and Gutiérrez-Expósito, Daniel
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OOCYSTS , *TOXOPLASMA gondii , *SHEEP , *PREGNANCY outcomes , *PHENOTYPES , *PHENOTYPIC plasticity , *ABORTION , *WOUND healing - Abstract
Toxoplasma gondii is a major cause of reproductive failure in small ruminants. Genotypic diversity of T. gondii strains has been associated with variations in phenotypic traits in in vitro and murine models. However, whether such diversity could influence the outcome of infection in small ruminants remains mostly unexplored. Here, we investigate the outcome of oral challenge in sheep at mid-pregnancy with 10 sporulated oocysts from three different T. gondii isolates belonging to archetypal II and III and selected according to their genetic and phenotypic variations shown in previous studies. Seventy-three pregnant sheep were divided in four groups: G1 infected with TgShSp1 isolate (type II, ToxoDB#3), G2 with TgShSp16 isolate (type II, ToxoDB#3), G3 with TgShSp24 isolate (type III, ToxoDB#2) and G4 of uninfected control sheep. Two different approaches were carried out within this study: (i) the outcome for the pregnancy after infection (n = 33) and (ii) the lesions and parasite tropism and burden at 14 and 28 days post infection (dpi) (n = 40). The onset of hyperthermia and seroconversion occurred one and two days later, respectively in G1 when compared to G2 and G3. However, sheep that suffered from reproductive failure, either by abortion, foetal dead at the time of euthanasia or stillbirth were similar among infected groups (50%, 40% and 47%, respectively). Histological lesions in placentomes and foetal tissues from euthanized animals from the second approach were only detected at 28 dpi and mainly in G1. At 14 dpi, T. gondii -DNA was only detected in G1 in the 11% of the placentomes. However, at 28 dpi the frequency of detection in placentomes was higher in G1 (96%) than in G2 and G3 (7% and 47%, respectively) besides in foetuses was lower in G2 (20%) than in G1 and G3 (100% and 87%, respectively). Regarding late abortions, stillbirths, and lambs of G1, G2 and G3, the frequency of microscopic lesions was similar between groups (79%, 78% and 67%, respectively) whereas T. gondii -DNA was evidenced in 100%, 55% and 100%, respectively. These recently obtained T. gondii isolates led to similar reproductive losses but intra- and inter-genotype variations in the rise of hyperthermia, dynamics of antibodies, frequency of lesions and parasite detection and distribution. Thus, the different phenotypic traits of the isolates could influence the outcome of the infection and mechanisms responsible for it, and further investigations are warranted. [Display omitted] • In pregnant sheep, reproductive losses after infection with T. gondii oocysts were similar despite the isolate employed. • Clinical outcomes reveal inter- and intra-genotypic differences. • Histological severity of microscopic lesions does not correlate with reproductive failure. • Invasion of the placenta causes unacknowledged early vascular lesions at 28 dpi. • Previous in vitro phenotypic traits cannot be directly inferred to pregnant sheep model. [ABSTRACT FROM AUTHOR]
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- 2023
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32. Seroepidemiologic study on the prevalence of Toxoplasma gondii and Trichinella spp. infections in black bears (Ursus americanus) in Pennsylvania, USA.
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Dubey, Jitender P., Brown, Justin, Ternent, Mark, Verma, Shiv K., Hill, Dolores E., Cerqueira-Cézar, Camila K., Kwok, Oliver C.H., Calero-Bernal, Rafael, and Humphreys, Jan G.
- Subjects
- *
TOXOPLASMOSIS in animals , *EPIDEMIOLOGY , *SERODIAGNOSIS , *BEARS , *DISEASE prevalence , *TOXOPLASMA gondii , *TRICHINELLA , *DISEASES - Abstract
The protozoan Toxoplasma gondii and the metazoan Trichinella spp. infect virtually all warm-blooded animals, including birds, humans, livestock, and marine mammals. Both parasitic infections can cause serious illness in human beings and can be acquired by ingesting under-cooked meat harboring infective stages. Approximately 3500 black bears ( Ursus americanus ) are legally-harvested each year in Pennsylvania, USA during the November hunting season. Among animals found infected with T. gondii , the prevalence of T. gondii is the highest among black bears in the USA; however, little is currently known of epidemiology of toxoplasmosis in this host species. Serum samples were collected during the winters of 2015 and 2016 from adult female bears and their nursing cubs or yearlings while they were still in their dens. Additionally, archived sera from bear samples collected throughout the year, including hunter-harvested bears in November and trapped bears in the summer, were serologically tested. Antibodies to T. gondii were assayed by the modified agglutination test (MAT, cut-off 1:25) and antibodies to Trichinella spp. were assayed using a commercial enzyme-linked immunosorbent assay (ELISA). Overall, T. gondii antibodies were found in 87.6% (206/235) of adults, and 44.1% (30/68) of yearlings. In March 2015/2016 sampling, antibodies to T. gondii were found in 94% (30/32) adult female bears while in their den. Antibodies were detected in 5% (3/66) of the nursing cubs in the dens of these sows. One positive cub had a MAT titer of 1:160 and two were positive at the 1:25 dilution but not at 1:50. The adult females of these cubs had MAT titers ranging from 1:400 to 1:3200. Antibodies to Trichinella spp. were found in 3% (6/181) of adults and 3.6% (1/28) of yearlings; these 7 bears were also seropositive for T. gondii . No antibodies to Trichinella spp. were detected in the sera of 44 nursing cubs tested. The finding of T. gondii antibodies in only 3 of 66 cubs, and higher antibody titers in their respective sows indicates that the colostrally-acquired antibodies wane to undetectable levels by 8–10 weeks, while the cubs are still in the den. The results indicate that there is no transplacental transmission of T. gondii , that antibodies acquired from colostrum are largely undetectable by the time cubs emerge from the den, and nearly that 50% of bears acquire infection postnatally by 10 months of age. This is the first report of disappearance of transcolostral antibodies of any infection in bears. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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