Preliminary genetic parameter estimates of meat quality traits in Hu sheep.

Because substantial numbers of Chinese consumers are prepared to pay for tender and quality lamb, meat quality traits are becoming more relevant for breeding programs for Chinese sheep breeds. The current study estimated heritabilities and genetic correlations for 13 meat quality traits recorded on lamb loins from Hu sheep. Heritability estimates ranged from 0.04 ± 0.06 for meat redness at 45 min to 0.57 ± 0.10 for drip loss, with most of the meat quality traits having moderate heritabilities. Positive genetic correlations were observed among meat color traits. Intramuscular fat (IMF) was genetically correlated with most meat quality traits, indicating that increasing IMF can favor meat pH, color, and tenderness, but would lead to increased cooking loss. Direct selection to increase IMF of loins is recommended to be included in breeding programs for Hu sheep, as it was more efficient than indirect selection on the other meat quality traits. The genetic parameters presented in this preliminary study provide valuable genetic information needed to design a breeding program aimed at improving the quality of lamb meat from Hu sheep.

Transcriptomic analysis of the HPG axis-related tissues reveals potential candidate genes and regulatory pathways associated with testicular size in Hu sheep.

Testicular size is an excellent proxy for selecting high-fertility rams. The hypothalamus-pituitary-gonadal (HPG) axis plays an important role in regulating reproductive capacity in vertebrates, while key genes and regulatory pathways within the HPG axis associated with testicular size remain largely unknown in sheep. This study comprehensively compared the transcriptomic profiles in the hypothalamus, pituitary and testis of rams after sexual maturity between the large-testis group (LTG, testicular weight = 454.29 ± 54.24 g) and the small-testis group (STG, testicular weight = 77.29 ± 10.76 g). In total, 914, 795 and 10518 differentially expressed genes (DEGs) were identified in the hypothalamus, pituitary and testis between LTG and STG, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that these DEGs were mainly involved in the biological processes of reproduction, biological regulation, and development process. Notably, the neuroactive ligand-receptor interaction and cAMP signaling pathways, commonly enriched by the DEGs in the hypothalamus and pituitary between two groups, were considered as two key signal pathways regulating testicular development through the HPGs axis. Weighted gene co-expression network analysis (WGCNA) identified two modules that were significantly associated with testicular size, and 97 key genes were selected with high module membership (MM) and gene significance (GS) in these two modules. Finally, a protein-protein interaction (PPI) network was constructed, and ten genes with the highest degree were represented as hub genes, including FOS, NPY, SST, F2, AGT, NTS, OXT, EDN1, VIP and TAC1. Taken together, these results provide new insights into the molecular mechanism underlying the HPG axis regulating testicular size of Hu sheep.

iTRAQ-based proteomic analysis provides novel insight into the postnatal testicular development of Hu sheep.

Testicular development is an intricate and coordinated process in which thousands of proteins are involved in the regulation of somatic cells development and spermatogenesis. However, knowledge about the proteomic changes during postnatal testicular development in Hu sheep is still elusive. The study was conducted to characterize the protein profiles at four key stages during postnatal testicular development, including infant (0-month-old, M0), puberty (3-month-old, M3), sexual maturity (6-month-old, M6) and body maturity (12-month-old, M12), and between the large- and small-testis groups at 6 months in Hu sheep. Consequently, 5252 proteins were identified using isobaric tags for relative and absolute quantification (iTRAQ) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods, and 465, 1261, 231 and 1080 differentially abundant proteins (DAPs) were found between M0_vs_M3, M3_vs_M6L, M6L_vs_M12, and M6L_vs_M6S, respectively. GO and KEGG analysis revealed that the majority of DAPs were involved in cellular process, metabolic process and immune system-related pathways. Furthermore, a protein-protein interaction network was constructed using 86 fertility-related DAPs, and five proteins with the highest degree were represented as hub proteins, including CTNNB1, ADAM2, ACR, HSPA2 and GRB2. This study provided new insights into the regulation mechanisms of postnatal testicular development and identified several potential biomarkers for selecting the high-fertility rams. SIGNIFICANCE OF THE STUDY: Testicular development is an intricate developmental process in which thousands of proteins are involved in regulating the somatic cells development and spermatogenesis. However, knowledge about the proteome changes during postnatal testicular development in Hu sheep is still elusive. This study provides comprehensive insights into the dynamic changes in the sheep testis proteome during postnatal testicular development. Additionally, testis size is positively correlated with semen quality and ejaculation volume, also for the merits of easy measurement, high heritability and selection efficiency, is an important indicator to select candidate rams with high fertility. The functional analyses of the acquired candidate proteins may help us gain a better understanding of the molecular regulatory mechanisms of testicular development.

Whole genome sequencing of simmental cattle for SNP and CNV discovery.

BACKGROUD: The single nucleotide polymorphisms (SNPs) and copy number variations (CNVs) are two major genomic variants, which play crucial roles in evolutionary and phenotypic diversity. RESULTS: In this study, we performed a comprehensive analysis to explore the genetic variations (SNPs and CNVs) of high sperm motility (HSM) and poor sperm motility (PSM) Simmental bulls using the high-coverage (25×) short-read next generation sequencing and single-molecule long reads sequencing data. A total of ~ 15 million SNPs and 2,944 CNV regions (CNVRs) were detected in Simmental bulls, and a set of positive selected genes (PSGs) and CNVRs were found to be overlapped with quantitative trait loci (QTLs) involving immunity, muscle development, reproduction, etc. In addition, we detected two new variants in LEPR, which may be related to the artificial breeding to improve important economic traits. Moreover, a set of genes and pathways functionally related to male fertility were identified. Remarkably, a CNV on SPAG16 (chr2:101,427,468 - 101,429,883) was completely deleted in all poor sperm motility (PSM) bulls and half of the bulls in high sperm motility (HSM), which may play a crucial role in the bull-fertility. CONCLUSIONS: In conclusion, this study provides a valuable genetic variation resource for the cattle breeding and selection programs.

Identification of Y-SNPs within ovine MSY region and their association with testicular size.

The screening of genomic variations within the male-specific region of the mammalian Y chromosome (MSY) is one of the most effective ways to investigate paternal evolutionary history and identify molecular markers related to male fertility. The current study was to identify single nucleotide polymorphisms (SNPs) within single-copy genes of the ovine MSY, and confirm whether they are associated with testicular size. A total of 21 Y-specific gene fragments were successfully amplified to screen Y-SNPs in 956 rams across nine sheep breeds. Three Y-SNPs, including SRY16: g.88 A > G in South African Mutton Merino sheep, ZFY16: g.146 C > T in Suffolk and South African Mutton Merino sheep, and EIF2S3Y2: g.77 C > G in Hu and Tan sheep, were identified using DNA-pooled sequencing and PCR restriction fragment length polymorphism (PCR-RFLP) methods. The investigation of the global distribution for three Y-SNPs showed that the C allele of ZFY16: g.146 C > T co-segregated with haplogroup y-HC, and the C/G allele of EIF2S3Y2: g.77 C > G co-segregated with haplogroup y-HA/y-HB1 in Hu sheep according to data mining from a previous study. In addition, association analysis revealed that ZFY16: g.146 C > T had a significant effect on yearling scrotal circumference in Suffolk sheep, and EIF2S3Y2: g.77 C > G was significantly associated with testicular and epididymis weight in Hu sheep (P ≤ 0.05). The current study concluded that Y-SNPs were associated with testicular size in specific sheep, which provides valuable candidate makers for selecting elite rams at an early age.

PRAMEY: A Bovid-Specific Y-Chromosome Multicopy Gene Is Highly Related to Postnatal Testicular Growth in Hu Sheep.

PRAMEY (preferentially expressed antigen in melanoma, Y-linked) belongs to the cancer-testis antigens (CTAs) gene family and is predominantly expressed in testis, playing important roles in spermatogenesis and testicular development. This study cloned the full-length cDNA sequence of ovine PRAMEY using the rapid amplification of cDNA ends (RACE) method and analyzed the expression profile and copy number variation (CNV) of PRAMEY using quantitative real-time PCR (qPCR). The results revealed that the PRAMEY cDNA was 2099 bp in length with an open reading frame (ORF) of 1536 bp encoding 511 amino acids. PRAMEY was predominantly expressed in the testis and significantly upregulated during postnatal testicular development. The median copy number (MCN) of PRAMEY was 4, varying from 2 to 25 in 710 rams across eight sheep breeds. There was no significant correlation between the CNV of PRAMEY and testicular size, while a significant positive correlation was observed between the mRNA expression and testicular size in Hu sheep. The current study suggests that the expression levels of PRAMEY were closely associated with testicular size, indicating that PRAMEY may play an important role in testicular growth.

Copy number variation of ZNF280BY across eight sheep breeds and its association with testicular size of Hu sheep.

ZNF280BY, a bovid-specific Y chromosome gene, was firstly found to be highly expressed in bovine testis, indicating that it may play important roles in testicular development and male fertility. In this study, we firstly cloned the full-length cDNA of ovine ZNF280BY containing 1,993 bp, and with a 1,632 bp open reading frame. ZNF280BY was predominantly expressed in the testis, and its expression level was significantly higher in large testis than in small testis in Hu sheep at 6 months of age. In addition, the expression level of ZNF280BY significantly increased during testicular development, showing the highest expression level at 12 mo of age. ZNF280BY showed copy number variation (CNV) in 723 rams from eight sheep breeds, ranging from 17 to 514 copies, with a median copy number of 188. Pearson correlation analysis showed that the CNV of ZNF280BY was negatively correlated with testis size in Hu sheep. Furthermore, its mRNA expression level in testis had no significant correlation with the CNV but was significantly correlated with testis size. This study concluded that the expression of ZNF280BY was closely related to testicular development, and the CNV of ZNF280BY could be used as an important genetic marker to evaluate the ram reproductive capacity at an early stage in Hu sheep.

The zinc finger protein 280B-like, Y-linked ZNF280BY is a bovid-specific Y chromosome gene, playing important roles in testicular development and male fertility. In this study, we found that the ovine ZNF280BY was highly expressed in the testis and significantly upregulated during testicular development. The copy number of ZNF280BY was negatively correlated with testicular size in Hu sheep. Our results demonstrated that the CNV of ZNF280BY could be used as an important candidate marker for selecting elite rams at an early stage in Hu sheep.

Identification of Key Genes Related to Postnatal Testicular Development Based on Transcriptomic Data of Testis in Hu Sheep.

The selection of testis size can improve the reproductive capacity of livestock used for artificial insemination and has been considered as an important strategy for accelerating the breeding process. Although much work has been done to investigate the mechanisms of testis development in various species, there is little information available in regard to the differences in transcriptomic profiling of sheep testes at different developmental stages. In this work, we aimed to identify differentially expressed genes (DEGs) by RNA-Seq in sheep during different growth stages, including 0 month old (infant, M0), 3 months old (puberty, M3), 6 months old (sexual maturity, M6) and 12 months old (body maturity, M12). A total of 4,606 (2,381 up and 2,225 down), 7,500 (4,368 up and 3,132 down), 15 (8 up and seven down) DEGs were identified in M3_vs_M0, M6_vs_M3, and M12_vs_M6 comparison, respectively. Of which, a number of genes were continuously up-regulated and down-regulated with testicular development, including ODF3, ZPBP1, PKDREJ, MYBL1, PDGFA, IGF1, LH, INSL3, VIM, AMH, INHBA, COL1A1, COL1A2, and INHA. GO analysis illustrated that DEGs were mainly involved in testis development and spermatogenesis. KEGG analysis identified several important pathways and verified several reproduction-associated DEGs such as COL1A1, COL1A2, PDGFA, and IGF1. In addition, two gene modules highly associated with testis development and core genes with testis size were identified using weighted gene co-expression network analysis (WGCNA), including hub genes positively associated with testis size such as RANBP9, DNAH17, SPATA4, CIB4 and SPEM1, and those negatively associated with testis size such as CD81, CSK, PDGFA, VIM, and INHBA. This study comprehensively identified key genes related to sheep testicular development, which may provide potential insights for understanding male fertility and better guide in animal breeding.

Influence of graphene oxide nanosheets on the cotransport of cu-tetracycline multi-pollutants in saturated porous media.

Antibiotic-heavy metal multi-pollutants are produced by intensive livestock farming and become an increasingly prominent problem. In this study, the transport behavior of tetracycline (TC) and its chelate with copper ions (Cu-TC) in saturated sand column with and without graphene oxide (GO) prefilled was investigated by laboratory breakthrough experiments. The effects of pH, ionic strength, and the cotransport with GO were studied detailedly. The results showed that the prepared nano-GO had a single- or multilayered sheet structure with a diameter of several µm. The surface of GO contained abundant oxygen-containing functional groups, which imparted it strong hydrophilicity and electronegativity. Pollutant transport experiments showed that decrease of H+ weakened the transport ability of TC and Cu-TC. Both Na+ and Ca2+ promoted the transport of TC, with Ca2+ having a much greater effect. The presence of Na+ inhibited the transport of Cu-TC, while Ca2+ promoted Cu-TC transport. The addition of Cu2+ was more favorable for the transport of Cu-TC than TC alone. In the GO-prefilled column, the effluent concentrations of TC and Cu-TC greatly decreased due to adsorption onto GO surfaces. The transport of Cu-TC was more related to GO concentration than TC alone due to the high affinity between GO and Cu-TC. Moreover, the transport behavior of GO in the sand column was consistent with that of the corresponding TC or Cu-TC, indicating that GO could cotransport with TC and Cu-TC multi-pollutants. Our study showed that the GO would interact with TC and Cu-TC and thus have significant influences on the fate and transport of these pollutions in porous media.

Flow regime identification for air valves failure evaluation in water pipelines using pressure data.

Air valve failure can cause air accumulation and result in a loss of carrying capacity, pipe vibration and even in some situations a catastrophic failure of water transmission pipelines. Air is most likely to accumulate in downward sloping pipes, leading to flow regime transition in these pipes. The flow regime identification can be used for fault diagnosis of air valves, but has received little attention in previous research. This paper develops a flow regime identification method that is based on support vector machines (SVMs) to evaluate the operational state of air valves in freshwater/potable pipelines using pressure signals. The laboratory experiments are set up to collect pressure data with respect to the four common flow regimes: bubbly flow, plug flow, blow-back flow and stratified flow. Two SVMs are constructed to identify bubbly and plug flows and validated based on the collected pressure data. The results demonstrate that pressure signals can be used for identifying flow regimes that represent the operational state (functioning or malfunctioning) of air valves. Among several signal features, Power Spectral Density and Short-Zero Crossing Rate are found to be the best indictors to classify flow regimes by SVMs. The sampling rate and time of pressure signals have significant influence on the performance of SVM classification. With optimal SVM features and pressure sampling parameters the identification accuracies exceeded 93% in the test cases. The findings of this study show that the SVM flow regime identification is a promising methodology for fault diagnosis of air valve failure in water pipelines.