Porcine epidemic diarrhea (PED) is a highly contagious, enteric disease of swine caused by the porcine epidemic diarrhea virus (PEDV). To find a suitable ELISA method to assess the infection of PEDV and the effectiveness of vaccines, we developed and evaluated an indirect enzyme-linked immunosorbent assay (iELISA) based on a truncated recombinant spike (S) protein expressed in Escherichia coli. The parameters of the iELISA were optimized, and the cutoff value determined as 0.259 by analyzing optical density (OD) values of 80 PEDV negative sera confirmed by western blot. Repeatability tests revealed that the coefficients of variation of positive sera within and between runs were both less than 10%. Cross-reactivity assays demonstrated that iELISA was PEDV-specific. A virus neutralization test with sera of 7 different OD values showed a positive correlation between the OD values and virus neutralization. The results suggest this iELISA is specific, sensitive, and repeatable. Further studies should focus on the relationship between OD values of sera and its virus neutralization.
Background Grasshoppers are typical phytophagous pests, and they have large appetites with high utilization of plants fibers, the digestion of which may depend on the microorganisms in their intestines. Grasshoppers have the potential to be utilized in bioreactors, which could improve straw utilization efficiency in the future. In this study, we describe the gut microbiome in three species of grasshoppers, Oedaleus decorus asiaticus, Aiolopus tamulus and Shirakiacris shirakii, by constructing a 16S rDNA gene library and analyzed the digestibility of cellulose and hemicellulose in the grasshoppers by using moss black phenol colorimetry and anthrone colorimetry. Results There were 509,436 bacterial OTUs (Operational Taxonomic Units) detected in the guts of all the grasshoppers sampled. Among them, Proteobacteria and Firmicutes were the most common, Aiolopus tamulus had the highest bacterial diversity, and Shirakiacris shirakii had the highest bacterial species richness. The intestinal microflora structure varied between the different species of grasshopper, with Aiolopus tamulus and Shirakiacris shirakii being the most similar. Meanwhile, the time at which grasshopper specimens were collected also led to changes in the intestinal microflora structure in the same species of grasshoppers. Klebsiella may form the core elements of the microflora in the grasshopper intestinal tract. The digestibility of cellulose/hemicellulose among the three species grasshoppers varied (38.01/24.99%, 43.95/17.21% and 44.12/47.62%). LEfSe analysis and Spearman correlation coefficients showed that the hemicellulosic digestibility of Shirakiacris shirakii was significantly higher than that of the other two species of grasshopper, which may be related to the presence of Pseudomonas, Stenotrophomonas, Glutamicibacter, Corynebacterium, and Brachybacterium in Shirakiacris shirakii intestinal tract. Conclusion The intestinal microbial communities of the three grasshoppers species are similar on phylum level, but the dominant genera of different species grasshoppers are different. The cellulose digestibility of the three species of grasshoppers is relatively high, which may be correlated with the presence of some gut microbiome. Increasing the understanding of the structure and function of the grasshopper intestinal microflora will facilitate further research and the utilization of intestinal microorganisms in the future.
Haemophilus parasuis is the etiological agent of Glässer's disease in pigs and 15 standard serovars were identified. The widespread disease causes great economic loss in the swine industry worldwide. Aiming to investigate the differences in genome composition and functions among various strains, a highly virulent strain ZJ0906 of H. parasuis serovar 12 from China was analyzed and compared with serovar 5 SH0165. Strain ZJ0906 genome is 2,324,740 base pairs with 40.06% genomic GC content. It contains 2,484 open reading frames (ORF) predicted by Glimmer 3.02, of which 2,352 (∼94.7%) were annotated by NCBI nr blast, 1,745 by COG database and 1,829 by KEGG database. 109 potential virulence factors were annotated in strain ZJ0906 and 3 of which are potentially related to antibiotic resistance. Strain ZJ0906 genome is ∼55 kilobases longer than SH0165 genome, with an extra 211 predicted ORFs. VFDB, ARDB, and PAIDB blast searches showed that ZJ0906 and SH0165 shared a nearly identical panel of potential virulence factors, drug resistant genes and four PAI-like regions which showed high homology to Enterococcus, Escherichia and Salmonella. Synteny analysis showed that gene rearrangements are frequent between the two strains, which may lead to variations in pathogenicity and cross-protection among serovars. KEGG pathway analyses showed strain ZJ0906 shared similar metabolic pathways to strain SH0165. Molecular identification of these genomic elements and potential virulence factors pave the way to the better understanding of mechanisms underlying metabolic capabilities and pathogenicity of H. parasuis and prospective vaccine targets besides the widely used method of inactivated bacteria.
Mitogenomes have been widely used for exploring phylogenetic analysis and taxonomic diagnosis. In this study, the complete mitogenomes of five species of Filchnerella were sequenced, annotated and analyzed. Then, combined with other seven mitogenomes of Filchnerella and four of Pamphagidae, the phylogenetic relationships were reconstructed by maximum likelihood (ML) and Bayesian (BI) methods based on PCGs+rRNAs. The sizes of the five complete mitogenomes are Filchnerella sunanensis 15,656 bp, Filchnerella amplivertica 15,657 bp, Filchnerella nigritibia 15,661 bp, Filchnerella pamphagoides 15,661 bp and Filchnerella dingxiensis 15,666 bp. The nucleotide composition of mitogenomes is biased toward A+T. All tRNAs could be folded into the typical clover-leaf structure, except that tRNA Ser (AGN) lacked a dihydrouridine (DHU) arm. The phylogenetic relationships of Filchnerella species based on mitogenome data revealed a general pattern of wing evolution from long wing to increasingly shortened wing.
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