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Background: Growing evidence indicates that N7-methylguanosine (m7G) modification plays critical roles in epigenetic regulation. However, no data regarding m7G modification are currently available in Eimeria tenella, a highly virulent species causing coccidiosis in chickens. Methods: In the present study, we explore the distribution of internal messenger RNA (mRNA) m7G modification in sporulated and unsporulated oocysts of E. tenella as well as its potential biological functions during oocyst development using methylated RNA immunoprecipitation sequencing (MeRIP-seq) and mRNA sequencing (mRNA-seq), and the mRNA-seq and MeRIP-seq data were verified by the quantitative reverse transcription polymerase chain reaction (RT–qPCR) and MeRIP–qPCR, respectively. Results: Our data showed that m7G peaks were detected throughout the whole mRNA body, and the coding DNA sequence (CDS) region displayed the most methylation modification. Compared with unsporulated oocysts, 7799 hypermethylated peaks and 1945 hypomethylated peaks were identified in sporulated oocysts. Further combined analysis of differentially methylated genes (DMGs) and differentially expressed genes (DEGs) showed that there was a generally positive correlation between m7G modification levels and gene transcript abundance. Unsurprisingly, the mRNA-seq and MeRIP-seq data showed good consistency with the results of the RT–qPCR and MeRIP–qPCR, respectively. Gene Ontology (GO) and pathway enrichment analysis of DEGs with altered m7G-methylated peaks were involved in diverse biological functions and pathways, including DNA replication, RNA transport, spliceosome, autophagy-yeast, and cAMP signaling pathway. Conclusions: Altogether, our findings revealed the potential significance of internal m7G modification in E. tenella oocysts, providing some directions and clues for later in-depth research. [ABSTRACT FROM AUTHOR]

Simple Summary: Trichomonads are among the most prevalent intestinal parasites with a worldwide distribution which can infect many animals, resulting in economic losses and threatening public health. The donkey raising industry in Shanxi Province is relatively well-developed; however, it is not yet known whether donkeys in Shanxi Province were infected with Tetratrichomonas buttreyi and Pentatrichomonas hominis. Thus, 815 fecal samples were collected from donkeys in three representative geographical locations in Shanxi Province to determine the prevalence and associated risk factors of T. buttreyi and P. hominis in donkeys using molecular approaches. The overall prevalence of T. buttreyi and P. hominis in donkeys in Shanxi Province was 25.4% and 0.7%, respectively. Genetic analysis revealed that all P. hominis sequences obtained in this study were identified as genotype CC1, suggesting possible zoonotic potential. This is the first report of T. buttreyi and P. hominis prevalence in donkeys worldwide, which not only extends the geographical distribution of trichomonads but also expands the host spectrum. The findings also have implications for the prevention and control of trichomonad infections in donkeys in Shanxi Province. Two species of trichomonads, Tetratrichomonas buttreyi and Pentatrichomonas hominis, are common intestinal parasites that can impact animal health and productivity. Severe infection by these parasites can lead to diarrhea and wasting in affected animals. Notably, P. hominis is known to cause diarrhea and has the potential to be transmitted between animals and humans. Donkeys hold significant economic importance in China's agricultural sector. However, whether donkeys are infected with T. buttreyi and P. hominis remains unknown globally. To address this gap in knowledge, 815 fecal samples were collected from donkeys in three representative regions in Shanxi Province, North China. Then, the presence and genetic characteristics of T. buttreyi and P. hominis were examined using species-specific PCR primers amplifying the small subunit ribosomal RNA genes. The overall prevalence was detected to be 25.4% (207/815) for T. buttreyi and 0.7% (6/815) for P. hominis in donkeys in Shanxi Province. All obtained P. hominis sequences were identified as genotype CC1. Genetic analysis revealed that all P. hominis isolates from donkeys were clustered into the same branch with isolates detected in humans, suggesting possible zoonotic transmission. This study is the first to report the occurrence and prevalence of T. buttreyi and P. hominis in donkeys globally. These findings expand the host range of trichomonads and improve our understanding of their genetic diversity and zoonotic potential, providing essential baseline data for the prevention and control of these parasites in donkeys in the region. [ABSTRACT FROM AUTHOR]

Simple Summary: Blastocystis spp. and Giardia duodenalis are two prevalent intestinal parasites with a worldwide distribution that can infect humans and animals, resulting in significant public health concerns and economic losses. For goats, infection with the two parasites can cause symptoms such as diarrhea, which is not conducive to the development of stockbreeding. Shanxi Province is one of the largest goats breeding provinces in China. However, the prevalence of Blastocystis spp. and G. duodenalis in black goats in Shanxi Province remains unknown. Thus, 1200 fecal samples of black goats were collected in five representative geographical locations in Shanxi Province to examine the presence and genotypes of G. duodenalis and Blastocystis spp. by using a molecular approach. The results showed that the total infection rates of G. duodenalis and Blastocystis spp. were 7.5% and 3.5%, respectively. The presence and genotypes of G. duodenalis were determined based on three established loci (tpi, bg, and gdh). Among the detected assemblages B and E of G. duodenalis, the most prevalent assemblage was E in black goats in the five study areas. One novel MLG (MLG-E12) was identified by multilocus genotypes (MLGs) analysis. Through DNA sequence analysis, four subtypes of Blastocystis spp. were found in black goats, namely ST5, ST10, ST14, and ST30, among which ST10 was the dominant subtype in this study. This is the first report of Blastocystis spp. and G. duodenalis infection in black goats in Shanxi Province, which not only enhances our understanding of the genetic diversity of Blastocystis spp. and G. duodenalis in black goats in China but also provides essential baseline data for the prevention and control of Blastocystis spp. and G. duodenalis infection in black goats in the study areas. Blastocystis spp. and Giardia duodenalis are two prevalent zoonotic intestinal parasites that can cause severe diarrhea and intestinal diseases in humans and many animals. Black goat (Capra hircus) farming is increasingly important in China due to the remarkable adaptability, high reproductive performance, rapid growth rate, and significant economic value of black goats. A number of studies have indicated that black goats are the potential reservoir of multiple zoonotic protozoans in China; however, the prevalence and zoonotic status of G. duodenalis and Blastocystis spp. in black goats in Shanxi Province is still unknown. Thus, a total of 1200 fecal samples of black goats were collected from several representative regions at different altitudes in Shanxi Province and were examined for the presence and genotypes of G. duodenallis and Blastocystis spp. by amplifying the beta-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) loci of G. duodenalis and SSU rRNA of Blastocystis spp. using PCR and sequence analysis methods, respectively. The overall prevalence of G. duodenalis and Blastocystis spp. in black goats in Shanxi Province were 7.5% and 3.5%, respectively. Two assemblages (B and E) of G. duodenalis and four subtypes (ST5, ST10, ST14, and ST30) of Blastocystis spp. were identified, with assemblage E and ST10 as the prevalent genotype and subtype in black goats, respectively. One novel multilocus genotype (MLG) was identified in MLG-E and was designated as MLG-E12. For both G. duodenalis and Blastocystis spp., the prevalence was significantly related to the region and age groups (p < 0.05). This is the first report on the prevalence of G. duodenalis and Blastocystis spp. in black goats in Shanxi Province. These results not only provide baseline data for the prevention and control of both parasites in black goats in Shanxi Province, but also enhance our understanding of the genetic composition and zoonotic potential of these two parasites. [ABSTRACT FROM AUTHOR]