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Mermithidae is a family of nematodes that parasitize a wide range of invertebrates worldwide. Herein, we report nematodes that were unexpectedly found in three of 486 adult stable flies (Stomoxys calcitrans) captured from three farms (F1, F2, and F3) in different regions of Gifu Prefecture, Japan. We aimed to characterize these nematodes both at the morphological and molecular level. Morphological studies revealed that the nematodes were juveniles of Mermithidae. Phylogenetic analysis based on 18S and 28S rDNA indicated that the mermithids from farms F1 and F2 could be categorized into the same cluster as Ovomermis sinensis and Hexamermis sp., whereas the mermithid from farm F3 clustered with Amphimermis sp. Additionally, these mermithids could be categorized within the same clusters as related mermithids detected in Japan that parasitize various arthropod orders. Our findings suggest that these stable flies may have been parasitized by mermithids already present in the region and that genetically distinct species of mermithids occur across Japan. To the best of our knowledge, this is the first report of mermithids parasitizing adult stable flies in Japan.
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Heterogeneity among both primed and naive pluripotent stem cell lines remains a major unresolved problem. Here we show that expressing the maternal-specific linker histone H1FOO fused to a destabilizing domain (H1FOO-DD), together with OCT4, SOX2, KLF4, and LMYC, in human somatic cells improves the quality of reprogramming to both primed and naive pluripotency. H1FOO-DD expression was associated with altered chromatin accessibility around pluripotency genes and with suppression of the innate immune response. Notably, H1FOO-DD generates naive induced pluripotent stem cells with lower variation in transcriptome and methylome among clones and a more uniform and superior differentiation potency. Furthermore, we elucidated that upregulation of FKBP1A, driven by these five factors, plays a key role in H1FOO-DD-mediated reprogramming.
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Reprogramación Celular , Histonas , Células Madre Pluripotentes Inducidas , Factor 4 Similar a Kruppel , Reprogramación Celular/genética , Humanos , Células Madre Pluripotentes Inducidas/citología , Células Madre Pluripotentes Inducidas/metabolismo , Histonas/metabolismo , Diferenciación Celular/genética , Factores de Transcripción de Tipo Kruppel/metabolismo , Factores de Transcripción de Tipo Kruppel/genética , Factores de Transcripción SOXB1/metabolismo , Factores de Transcripción SOXB1/genética , Cromatina/metabolismo , Células Madre Pluripotentes/metabolismo , Células Madre Pluripotentes/citología , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , TranscriptomaRESUMEN
Toxoplasma gondii is a global protozoan pathogen. Clonal lineages predominate in Europe, North America, Africa, and China, whereas highly recombinant parasites are endemic in South/Central America. Far East Asian T. gondii isolates are not included in current global population genetic structure analyses at WGS resolution. Here we report a genome-wide population study that compared eight Japanese and two Chinese isolates against representative worldwide T. gondii genomes using POPSICLE, a novel population structure analyzing software. Also included were 7 genomes resurrected from non-viable isolates by target enrichment sequencing. Visualization of the genome structure by POPSICLE shows a mixture of Chinese haplogroup (HG) 13 haploblocks introgressed within the genomes of Japanese HG2 and North American HG12. Furthermore, two ancestral lineages were identified in the Japanese strains; one lineage shares a common ancestor with HG11 found in both Japanese strains and North American HG12. The other ancestral lineage, found in T. gondii isolates from a small island in Japan, is admixed with genetically diversified South/Central American strains. Taken together, this study suggests multiple ancestral links between Far East Asian and American T. gondii strains and provides insight into the transmission history of this cosmopolitan organism.
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Genoma de Protozoos , Filogenia , Toxoplasma , Toxoplasma/genética , Toxoplasma/clasificación , Humanos , América del Norte , Genoma de Protozoos/genética , Toxoplasmosis/parasitología , China , América Central , Japón , Haplotipos , Variación Genética , Recombinación GenéticaRESUMEN
We report a confirmed case of Toxoplasma gondii infection in the lungs of a cow exhibiting respiratory symptoms. At slaughter, white nodules were discovered in lung tissue, accompanied by enlarged hilar lymph nodes. Histological examination revealed the disappearance of alveolar structures in nodular areas, replaced by granulomas containing inflammatory cells. Immunohistochemical staining with anti-T. gondii antibody and nucleotide sequencing of 18S rDNA confirmed T. gondii infection. However, the link between T. gondii and observed symptoms remains unclear. Various factors, including host genetics, underlying diseases, infection route, and exposure level, may contribute to these uncommon symptoms. Although T. gondii infections in cattle are traditionally considered asymptomatic, our study suggests the possible existence of clinical symptoms associated with Toxoplasma infection. Beef cattle are generally not assumed to be a relevant source of human T. gondii infection; however, sporadic transmission by infected edible beef to humans cannot be completely excluded and deserves further studies.
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Enfermedades de los Bovinos , Toxoplasma , Toxoplasmosis Animal , Bovinos , Toxoplasma/aislamiento & purificación , Toxoplasma/genética , Animales , Toxoplasmosis Animal/parasitología , Toxoplasmosis Animal/patología , Toxoplasmosis Animal/diagnóstico , Enfermedades de los Bovinos/parasitología , Enfermedades de los Bovinos/patología , Pulmón/parasitología , Pulmón/patología , Neumonía/parasitología , Neumonía/veterinaria , Femenino , Granuloma/parasitología , Granuloma/patología , ARN Ribosómico 18S/análisisRESUMEN
The intervillous space of human placenta is filled with maternal blood, and villous trophoblasts are constantly exposed to the shear stress generated by maternal blood pressure and flow throughout the entire gestation period. However, the effects of shear stress on villous trophoblasts and their biological significance remain unknown. Here, using our recently established naïve human pluripotent stem cells-derived cytotrophoblast stem cells (nCTs) and a device that can apply arbitrary shear stress to cells, we investigated the impact of shear stress on early-stage trophoblasts. After 72 h of exposure to 10 dyn/cm2 shear stress, nCTs became fused and multinuclear, and mRNA expression of the syncytiotrophoblast (ST) markers, such as glial cell missing 1, endogenous retrovirus group W member 1 envelope, chorionic gonadotropin subunit beta 3, syndecan 1, pregnancy specific beta-1-glycoprotein 3, placental growth factor, and solute carrier family 2 member 1 were significantly upregulated compared to static conditions. Immunohistochemistry showed that shear stress increased fusion index, human chorionic gonadotropin secretion, and human placental lactogen secretion. Increased microvilli formation on the surface of nCTs under flow conditions was detected using scanning electron microscopy. Intracellular cyclic adenosine monophosphate significantly increased under flow conditions. Moreover, transcriptome analysis of nCTs subjected to shear stress revealed that shear stress upregulated ST-specific genes and downregulated CT-specific genes. Collectively, these findings indicate that shear stress promotes the differentiation of nCTs into ST.
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Células Madre Pluripotentes Inducidas , Placenta , Femenino , Embarazo , Humanos , Placenta/metabolismo , Células Madre Pluripotentes Inducidas/metabolismo , Factor de Crecimiento Placentario/metabolismo , Trofoblastos/metabolismo , Gonadotropina Coriónica/farmacología , Gonadotropina Coriónica/metabolismo , Diferenciación CelularRESUMEN
Recently, several studies using cultures of human embryos together with single-cell RNA-seq analyses have revealed differences between humans and mice, necessitating the study of human embryos1-8. Despite the importance of human embryology, ethical and legal restrictions have limited post-implantation-stage studies. Thus, recent efforts have focused on developing in vitro self-organizing models using human stem cells9-17. Here, we report genetic and non-genetic approaches to generate authentic hypoblast cells (naive hPSC-derived hypoblast-like cells (nHyCs))-known to give rise to one of the two extraembryonic tissues essential for embryonic development-from naive human pluripotent stem cells (hPSCs). Our nHyCs spontaneously assemble with naive hPSCs to form a three-dimensional bilaminar structure (bilaminoids) with a pro-amniotic-like cavity. In the presence of additional naive hPSC-derived analogues of the second extraembryonic tissue, the trophectoderm, the efficiency of bilaminoid formation increases from 20% to 40%, and the epiblast within the bilaminoids continues to develop in response to trophectoderm-secreted IL-6. Furthermore, we show that bilaminoids robustly recapitulate the patterning of the anterior-posterior axis and the formation of cells reflecting the pregastrula stage, the emergence of which can be shaped by genetically manipulating the DKK1/OTX2 hypoblast-like domain. We have therefore successfully modelled and identified the mechanisms by which the two extraembryonic tissues efficiently guide the stage-specific growth and progression of the epiblast as it establishes the post-implantation landmarks of human embryogenesis.
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Desarrollo Embrionario , Estratos Germinativos , Células Madre Pluripotentes , Humanos , Diferenciación Celular , Implantación del Embrión , Embrión de Mamíferos/citología , Embrión de Mamíferos/embriología , Embrión de Mamíferos/metabolismo , Desarrollo Embrionario/genética , Desarrollo Embrionario/fisiología , Estratos Germinativos/citología , Estratos Germinativos/embriología , Estratos Germinativos/metabolismo , Células Madre Pluripotentes/citología , Interleucina-6/metabolismo , Gástrula/citología , Gástrula/embriología , Amnios/citología , Amnios/embriología , Amnios/metabolismo , Ectodermo/citología , Ectodermo/embriología , Ectodermo/metabolismo , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Factores de Transcripción Otx/genética , Factores de Transcripción Otx/metabolismoRESUMEN
In 2021, we showed that naive human pluripotent stem cells (PSCs) can differentiate into trophoblasts via trophectoderm (TE)-like cells. Since TE is a pre-implantation stage of trophoblasts constituting blastocysts, naive human PSCs are an invaluable tool for understanding the entire process of trophoblast development. It has been reported for many years that primed human PSCs can also differentiate into the trophoblast lineage. The in vitro differentiation of naive and primed human PSCs hints at the possibility that human pre- and even post-implantation epiblasts retain the differentiation potential into the trophoblast lineages in vivo. Here, we review the in vitro specification of trophoblasts from human PSCs. Moreover, we discuss the different trophoblast differentiation pathways from naive and primed PSCs.
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Células Madre Pluripotentes , Trofoblastos , Humanos , Trofoblastos/metabolismo , Diferenciación Celular/genética , Implantación del Embrión , Estratos GerminativosRESUMEN
Many parasite species migrate to another site of infection after entering the host body. Such parasite dynamics are closely related to pathogenicity, but it is not easy to observe such parasite behavior deep within the organs. In recent years, technology that can make organs transparent has been developed that enables us to observe deep within organs ex vivo while maintaining their three-dimensional structure. This review describes a series of attempts to apply this technology to understand the behavior of Toxoplasma gondii in the host body. A series of studies has shown that T. gondii tachyzoites that infect leukocytes can reach target organs far from the site of invasion via the circulatory system. In addition, infected leukocytes in the bloodstream adhere more readily to vascular endothelial cells than uninfected leukocytes and are more likely to remain inside the target organs. When infected leukocytes adhere to the vascular endothelial cells of the target organ, the tachyzoites inside the cells immediately escape and infect the parenchyma of the organs. As described above, organ transparency technology is a powerful tool for understanding the internal dynamics of parasites.
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Parásitos , Toxoplasma , Animales , Células Endoteliales/parasitología , LeucocitosRESUMEN
Primate germ cell development remains largely unexplored due to limitations in sample collection and the long duration of development. In mice, primordial germ cell-like cells (PGCLCs) derived from pluripotent stem cells (PSCs) can develop into functional gametes by in vitro culture or in vivo transplantation. Such PGCLC-mediated induction of mature gametes in primates is highly useful for understanding human germ cell development. Since marmosets generate functional sperm earlier than other species, recapitulating the whole male germ cell development process is technically more feasible. Here, we induced the differentiation of iPSCs into gonocyte-like cells via PGCLCs in marmosets. First, we developed an mRNA transfection-based method to efficiently generate PGCLCs. Subsequently, to promote PGCLC differentiation, xenoreconstituted testes (xrtestes) were generated in the mouse kidney capsule. PGCLCs show progressive DNA demethylation and stepwise expression of developmental marker genes. This study provides an efficient platform for the study of marmoset germ cell development.
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Callithrix , Semen , Humanos , Masculino , Animales , Ratones , Células Germinativas , Diferenciación Celular/genética , ARN Mensajero/genéticaRESUMEN
Reconstitution of the germ cell lineage using pluripotent stem cells provides a unique platform to deepen our understanding of the mechanisms underlying germ cell development and to produce functional gametes for reproduction. This study aimed to establish a culture system that induces a robust number of primordial germ cell-like cells (PGCLCs) from common marmoset (Callithrix jacchus) embryonic stem cells. The robust induction was achieved by not only activation of the conserved PGC-inducing signals, WNT and BMP4, but also temporal inhibitions of WNT and retinoic acid signals, which prevent mesodermal and neural differentiation, respectively, during PGCLC differentiation. Many of the gene expression and differentiation properties of common marmoset PGCLCs were similar to those of human PGCLCs, making this culture system a reliable and useful primate model. Finally, we identified PDPN and KIT as surface marker proteins by which PGCLCs can be isolated from embryonic stem cells without genetic manipulation. This study will expand the opportunities for research on germ cell development and production of functional gametes to the common marmoset.
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Callithrix , Células Madre Pluripotentes , Animales , Diferenciación Celular , Células Madre Embrionarias , Células Germinativas/metabolismo , Tretinoina/farmacología , Tretinoina/metabolismoRESUMEN
Encyclometra japonica Yoshida and Ozaki, 1929 is a fluke that parasitizes the digestive tract of snakes. When the species was first reported, it was originally characterized on the basis of morphologic features, namely, the characteristics of the 2 testes located obliquely in the anterior-posterior direction, and the position of the ventral sucker one-fourth to one-third from the anterior extremity. Thereafter, more specimens with other morphologic variations were reported, and a new morphologic feature for species discrimination was proposed. However, the proposal is uncertain, and the criteria for morphologic discrimination between E. japonica and other Encyclometra species remain debatable. Thus, morphologically discriminable characteristics on the basis of a sufficient number of specimens are required, and molecular methods for species identification that allow for a more objective assessment need to be established. Therefore, in the present study, nucleotide sequences of 18S, 28S ribosomal deoxyribonucleic acid and cytochrome c oxidase subunit I (COI) of E. japonica were determined along with detailed morphologic observations for the first time. Key characteristics were observed in all specimens, whereas some morphologic variations were observed in the symmetry of the cecum. The COI sequences, which are known to be variable genetic regions, were identical among all specimens; therefore, all of them were considered the same species. This result clarifies the high morphologic variation in E. japonica. The 18S and 28S sequences were 99.78% and 99.11-99.19% similar to those of Encylometra colubrimurorum. The high degree of homology between these genes indicates that E. japonica and E. colubrimurorum are closely related but independent species. The present data will be used for the identification of E. japonica and to evaluate the relationships within the genus Encyclometra.
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Trematodos , Animales , Japón , Filogenia , Tracto GastrointestinalRESUMEN
Naive human induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with Sendai virus (SeV) vectors. However, only dermal fibroblasts have been successfully reprogrammed this way, and the process requires culture on feeder cells. Moreover, SeV vectors are highly persistent and inhibit subsequent differentiation of iPSCs. Here, we report a modified SeV vector system to generate transgene-free naive human iPSCs with superior differentiation potential. The modified method can be applied not only to fibroblasts but also to other somatic cell types. SeV vectors disappear quickly at early passages, and this approach enables the generation of naive iPSCs in a feeder-free culture. The naive iPSCs generated by this method show better differentiation to trilineage and extra-embryonic trophectoderm than those derived by conventional methods. This method can expand the application of iPSCs to research on early human development and regenerative medicine.
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Células Madre Pluripotentes Inducidas , Humanos , Reprogramación Celular/genética , Virus Sendai/genética , Vectores Genéticos , Diferenciación Celular/genéticaRESUMEN
Free-ranging cats are invasive species threatening biodiversity worldwide. They may also impose an environmental risk to humans and livestock through the transmission of zoonotic diseases. We investigated antibody levels against Toxoplasma gondii in free-ranging cats and black rats (definitive and representative intermediate hosts) by ELISA and determined their relationships with landscape environmental factors on Tokunoshima Island, Japan, the Natural World Heritage site. We found a higher seroprevalence (>70%) in both cats and black rats in landscapes where the cattle barn density was high. This was consistent with higher density of rats revealed in our trapping survey. The spatial scale of landscape factors affecting infection was broader in cats (1 km buffer radius) than in black rats (100 m buffer radius). Both cats and rats showed an increasing trend in optical density (OD) values with increasing body weight and landscape cattle barn density, suggesting that the antibody concentration increases as the chance of exposure to T. gondii in the environment increases. Thus, management actions to stop humans from feeding cats and to control rat populations without using cats are both necessary to reduce the human health risk as well as to conserve endangered species on the island.
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Some filoviruses such as ebolaviruses and marburgviruses, cause hemorrhagic fever in humans and nonhuman primates. Pigs are suggested to play a potential role in the filovirus ecology. We investigated the seroprevalence of filovirus infection in pigs in Ghana. Using a viral glycoprotein (GP)-based enzyme-linked immunosorbent assay, we detected filovirus-specific immunoglobulin G antibodies in 5 of 139 samples. These positive sera showed specificities to four different filovirus species. Particularly, two of the positive sera reacted to GPs of two African ebolaviruses (i.e., Ebola virus and Taï Forest virus) in Western blotting. Our results suggest that these Ghanaian pigs were exposed to multiple filoviruses and emphasize the importance of continuous monitoring of filovirus infection in pig populations in West African countries.
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Ebolavirus , Infecciones por Filoviridae , Fiebre Hemorrágica Ebola , Enfermedades de los Porcinos , Porcinos , Humanos , Animales , Ghana/epidemiología , Fiebre Hemorrágica Ebola/diagnóstico , Fiebre Hemorrágica Ebola/veterinaria , Estudios Seroepidemiológicos , Anticuerpos Antivirales , Infecciones por Filoviridae/veterinaria , Enfermedades de los Porcinos/epidemiologíaRESUMEN
In this issue of Cell Stem Cell, Pham et al. report that naive human pluripotent stem cells can be differentiated into extraembryonic mesoderm cells (EXMCs). EXMCs are maintained for up to 70 days, resemble human and monkey extraembryonic mesoderm, and provide a model to study EXMC specification and function.
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Mesodermo , Células Madre Pluripotentes , Diferenciación Celular , HumanosRESUMEN
T-type Ca2+ channels (T-channels), particularly Cav3.2 and Cav3.1 isoforms, are promising targets for treating various diseases including intractable pain. Given the potent inhibitory activity of pimozide, an antipsychotic, against T-channels, we conducted structure-activity relationship studies of pimozide derivatives, and identified several compounds including 3a, 3s, and 4 that had potency comparable to that of pimozide in inhibiting T-channels, but little binding affinity to dopamine D2 receptors. The introduction of a phenylbutyl group on the benzoimidazole nuclei of pimozide was considered a key structural modification to reduce the binding affinity to D2 receptors. Those pimozide derivatives potently suppressed T-channel-dependent somatic and visceral pain in mice, without causing any motor dysfunctions attributable to D2 receptor blockade, including catalepsy. The present study thus provides an avenue to develop novel selective T-channel inhibitors available for pain management via the structural modification of existing medicines.
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Canales de Calcio Tipo T , Dolor Visceral , Ratones , Animales , Pimozida/farmacología , Pimozida/uso terapéutico , Canales de Calcio Tipo T/metabolismo , Dolor Visceral/tratamiento farmacológico , Dopamina , Bloqueadores de los Canales de Calcio/química , Receptores Dopaminérgicos/metabolismoRESUMEN
Porcine reproductive and respiratory syndrome (PRRS) is an infectious swine disease caused by the PRRS virus (PRRSV) that results in economic loss to the pig-rearing industry. To study PRRSV infection in wild boars and pigs, we conducted a serological survey in Gifu Prefecture, Japan, from 2020 to 2021. Three out of 453 (0.7%) wild boar sera were positive for PRRSV antibodies in a commercial ELISA. However, given that PRRSV RNA was not detected in these three wild boars and the specificity and sensitivity of the test kit, these are considered as false positives. Although seropositive pigs were found in multiple pig farms in the study area, the role of wild boars as a source of PRRS to pig farms appeared to be minimal.
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Síndrome Respiratorio y de la Reproducción Porcina , Virus del Síndrome Respiratorio y Reproductivo Porcino , Enfermedades de los Porcinos , Animales , Anticuerpos Antivirales , Ensayo de Inmunoadsorción Enzimática/veterinaria , Japón/epidemiología , Síndrome Respiratorio y de la Reproducción Porcina/epidemiología , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Sus scrofa , PorcinosRESUMEN
Mesocestoides vogae is a cestode of the order Cyclophyllidea, and its second intermediate hosts are mammals, birds, amphibians, and reptiles. The parasite forms a tetrathyridium in the second intermediate host and multiplies asexually, sometimes to the point of filling the host's abdominal cavity. Proliferated tetrathyridium may cause lethal conditions in the host. During the asexual multiplication period, the scolex first replicates into two and then divides into two worms. In this study, to investigate the factors that promote the replication, tetrathyridia were cultured under various in vitro conditions. When several worms which already had two scolexes were cultured together, the division into two worm bodies was complete, but when single worm was cultured, the division hardly proceeded. The result indicates that the division progression of tetrathyridia with two scolexes requires the presence of other worms. In contrast, tetrathyridia with only one scolex did not initiate the division process, whether cultured together or alone. Then, the necessity of direct contact between the bodies of the worms to promote the division of tetrathyridia with two scolexes was assessed. For this, the well of the culture plate was partitioned into upper and lower parts using a mesh, and 20 worms in the upper part and single worm in the lower part were cultured. In all examined wells, worms in the upper part showed complete division, whereas the worms in the lower part rarely completed the division. Thus, direct contact between tetrathyridium promotes the division of tetrathyridia bearing two scolexes.
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Infecciones por Cestodos , Mesocestoides , Animales , Infecciones por Cestodos/parasitología , Infecciones por Cestodos/veterinaria , MamíferosRESUMEN
Toxoplasmosis is a widespread protozoan zoonosis. Since ingesting undercooked meat harboring Toxoplasma gondii cyst is considered one of the major transmission routes to humans, the screening of T. gondii in meat-producing animals can reduce the risk of food-borne toxoplasmosis in humans. Among serological diagnostic methods, Luciferase-linked Antibody Capture Assay (LACA) has been found to be a promising platform with high sensitivity and specificity. In this study, we aimed to evaluate recombinant nanoluciferase fused-T. gondii antigens (rNluc-GRA6, rNluc-GRA7, rNluc-GRA8 and rNluc-BAG1) for their potential use in LACA for pigs. As a result, the sensitivity of GRA6-, GRA7-, GRA8- and BAG1-LACA were 70.0%, 80.0%, 80.0% and 30.0% with specificity 87.0%, 81.5%, 74.1% and 50.0%, respectively. The cocktail LACA using a mixture of rNluc-GRA6, rNluc-GRA7 and rNluc-GRA8 indicated higher sensitivity (90.0%) and a similar specificity (96.3%) in comparison with the commercial ELISA kit. Compared to the Dye-Test as a reference test, cocktail LACA showed strong agreement (kappa value=0.811) when we assessed pig sera collected at the slaughterhouse. In addition, we also successfully established the rapid LACA format for the detection of Toxoplasma infection in pigs (called Rapid-LACA) in which the test could be performed within 30 min. In Rapid-LACA, the protein A pre-coated/blocked plates could be preserved at -30°C, 4°C or room temperature conditions for at least two months without compromising on the quality of assay.