Genomic insights into the phylogeny and biomass-degrading enzymes of rumen ciliates.

Understanding the biodiversity and genetics of gut microbiomes has important implications for host physiology and industrial enzymes, whereas most studies have been focused on bacteria and archaea, and to a lesser extent on fungi and viruses. One group, still underexplored and elusive, is ciliated protozoa, despite its importance in shaping microbiota populations. Integrating single-cell sequencing and an assembly-and-identification pipeline, we acquired 52 high-quality ciliate genomes of 22 rumen morphospecies from 11 abundant morphogenera. With these genomes, we resolved the taxonomic and phylogenetic framework that revised the 22 morphospecies into 19 species spanning 13 genera and reassigned the genus Dasytricha from Isotrichidae to a new family Dasytrichidae. Comparative genomic analyses revealed that extensive horizontal gene transfers and gene family expansion provided rumen ciliate species with a broad array of carbohydrate-active enzymes (CAZymes) to degrade all major kinds of plant and microbial carbohydrates. In particular, the genomes of Diplodiniinae and Ophryoscolecinae species encode as many CAZymes as gut fungi, and ~80% of their degradative CAZymes act on plant cell-wall. The activities of horizontally transferred cellulase and xylanase of ciliates were experimentally verified and were 2-9 folds higher than those of the inferred corresponding bacterial donors. Additionally, the new ciliate dataset greatly facilitated rumen metagenomic analyses by allowing ~12% of the metagenomic sequencing reads to be classified as ciliate sequences.

Metagenomic analysis reveals antibiotic resistance genes in the bovine rumen.

Metagenomics and network analysis were used to profile antibiotic resistance genes (ARGs) and their cooccurrence patterns in bovine rumen microbes. A total of 4941 ruminal microbial genomes and 20 metagenome samples were used in this study. In general, 103 ARG subtypes belonging to 20 ARG types in 79 candidate genomes were identified, showing the broad-spectrum profiles of ARGs in the bovine rumen environment. A wide distribution of genes encoding bacitracin resistance was found among the candidate genomes, suggesting the possibility that bovines might be one of the sources of bacitracin resistance genes. Cooccurrence patterns were found within or between the ARG types, and a positive correlation was found between some ARGs and bacteria, which revealed potential dominant hosts of ARGs. The investigation showed that bovine rumen systems are important ARG reservoirs, and our research might provide a theoretical basis for the evaluation of the harmfulness of ARGs and antibiotic-resistant bacteria (ARB) to food safety and human health.

Gene co-expression network analysis reveals key potential gene modules in utero-vaginal junction associated with duration of fertility trait of breeder hens.

The number of days (DN) when hens lay fertile eggs as well as the number of fertile eggs (FN) were produced after a single artificial insemination (AI), including the two duration of fertility (DF) traits. Indeed, they are the key production performance that associates with the production cost of hatching egg when its determination the interval between successive artificial inseminations. However, the relevant genes response for regulating the DF has not been uncovered yet. Therefore, we performed a weighted gene co-expression network analysis (WGCNA) to investigate the insight into co-expression gene modules on DF process in hens. The total mRNA was extracted from the utero-vaginal junction (UVJ, with the sperm storage function in hen's oviduct which is the biological basis for DF) of 20 hens with several levels of DF traits, and performed transcriptome sequences of mRNA. As a result, three co-expression gene modules were identified to be highly correlated with DF traits. Moreover, the expression changes of top 5 hub genes in each module with DF traits were further confirmed in other 20 hens by RT-PCR. These findings highlighted the co-expression modules and their affiliated genes as playing important roles in the regulation of DF traits.

A transcriptomic comparison of theca and granulosa cells in chicken and cattle follicles reveals ESR2 as a potential regulator of CYP19A1 expression in the theca cells of chicken follicles.

Previous studies have shown that theca and granulosa cell layers in follicles do not play the same roles in mammals and birds, especially regarding the synthesis of estrogen. The functions of these two cell types have been well characterized in cattle, but they remain unclear in chickens. To clarify this issue, a comparison of small yellow follicles (SYFs) in chickens and cattle at different follicular development stages was done by weighted gene co-expression network analysis (WGCNA). The modules obtained from WGCNA were used for further identification of the key genes associated with CYP19A1 expression. Module preservation analysis showed high similarity between cow_D (the follicular phase before the LH surge) and chicken_SYF (small yellow follicle between 6 and 8 mm in diameter) datasets, and 10 top hub genes highly associated with CYP19A1 expression in chicken SYFs were identified in each module. A comparison of the transcriptomes of theca and granulosa cells (TCs and GCs) between chicken SYFs and cattle follicles at the differentiation stage, as well as the aforementioned hub genes, revealed that ESR2 is a potential regulator of CYP19A1 expression in the theca cells of chicken SYFs. Furthermore, 197 cell-specific (179 in theca and 18 in granulosa) and 235 cell-biased expressed genes (196 in theca and 39 in granulosa) in chicken small yellow follicles were also identified by transcriptomic comparison of theca and granulosa cells.