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Determining sensitive biomarkers in the peripheral blood to identify interstitial lung abnormalities (ILAs) is essential for the simple early diagnosis of ILAs. This study aimed to determine serum metabolic biomarkers of ILAs and the corresponding pathogenesis. Three groups of subjects undergoing health screening, including healthy subjects, subjects with ILAs, and subjects who were healthy initially and with ILAs one year later (Healthy→ILAs), were recruited for this study. The metabolic profiles of all of the subjects’ serum were analyzed by liquid chromatography quadruple time-of-flight mass spectrometry. The metabolic characteristics of the ILAs subjects were discovered, and the corresponding biomarkers were predicted. The metabolomic data from the Healthy→ILAs subjects were collected for further verification. The results indicated that five serum metabolite alterations (up-regulated phosphatidylcholine, phosphatidic acid, betaine aldehyde and phosphatidylethanolamine, as well as down-regulated 1-acylglycerophosphocholine) were sensitive and reliable biomarkers for identifying ILAs. Perturbation of the corresponding biological pathways (RhoA signaling, mTOR/P70S6K signaling and phospholipase C signaling) might be at least partially responsible for the pathogenesis of ILAs. This study may provide a good template for determining the early diagnostic markers of subclinical disease status and for obtaining a better understanding of their pathogenesis.

Currently Birnaviridae family contains four genera with all members identified from birds, fishes and insects only. The present study reports a novel birnavirus unexpectedly identified from classical swine fever virus (CSFV)-infected pigs by viral metagenomic analysis, therefore named as porcine birnavirus (PBRV). Follow-up RT-PCR screening of archived tissues of diseased pigs identified 16 PBRV strains from nine provinces/autonomous regions in China spanning 21 years (1998-2019), and the viral loads of PBRV in clinical samples were 105.°8-107.95 genome copies per 0.1 g tissue, showing the replication of PBRVs in the pigs. Genome-based sequence comparison showed that PBRVs are genetically distant from existing members within the Birnaviridae family with 45.8-61.6% and 46.2-63.2% nucleotide sequence similarities in segments A and B respectively, and the relatively closed viruses are avibirnavirus strains. In addition, indels of 57, 5, and 18 amino acid residues occurred in 16, 2, and 7 locations of the PBRV polyprotein, VP5, and VP1 proteins respectively as compared to the reference avibirnaviruses. Phylogenetic analysis showed that PBRVs formed an independent genotype separated from other 4 genera, which could be classified into 2 or 3 subgenotypes (PBRV-A1-2 and PBRV-B1-3) based on the nucleotide sequences of full preVP2 and VP1 genes respectively. All results showed that PBRV is a novel pig virus species, which constitutes the first mammalian birnavirus taxon, thereby naming as Mambirnavirus genus is proposed.

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