Detection and differentiation of seven porcine respiratory pathogens using a multiplex ligation-dependent probe amplification assay.

Respiratory diseases due to viral or bacterial agents, either alone or in combination, cause substantial economic burdens to the swine industry worldwide. Rapid and reliable detection of causal pathogens is crucial for effective epidemiological surveillance and disease management. This research aimed to employ the multiplex ligation-dependent probe amplification (MLPA) assay for simultaneous detection of seven distinct pathogens causing respiratory problems in swine, porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), porcine respiratory coronavirus (PRCV), porcine circovirus type 2 (PCV2), Pasteurella multocida, Actinobacillus pleuropneumoniae, and Glässerella parasuis. The results indicated no probe cross-reactivity among the seven target agents with other swine pathogens. The detection limits ranged from 5 to 34 copies per assay for the target organisms. The MLPA assay was evaluated with 88 samples and compared to real-time or multiplex PCR for the target pathogens. The MLPA assay demonstrated high relative test sensitivities (100 %) and reasonable to good relative specificities at 62.5 %, 95.1 %, 86.8 %, and 97.6 % for PRRSV, P. multocida, G. parasuis, and PCV2, respectively, relative to comparator PCR assays. In 71 samples where MLPA and comparator PCR assays matched exactly, infections were detected in 64 samples (90.1 %), with PRRSV being the most commonly found virus and 50.7 % of the samples showing co-infection with two to five of the pathogens. This approach serves as a valuable tool for conducting differential diagnoses and epidemiological investigations of pathogen prevalence within swine populations.

Electrospun polylactic acid nanofiber film modified by silver oxide deposited on hemp fibers for antibacterial fruit packaging.

Bacterial and fungal contamination have become major factors in fruit spoilage and damage, posing a potential risk to human health. In this work, polylactic acid (PLA) nanofibers combined with Ag2O-hemp fibers for a good antimicrobial effect were developed and applied to antimicrobial fruit fresh-keeping packages. The results of molecular simulation calculations showed that the strength of hydrogen bonds between Ag2O and hemp fibers reached 45.522 kJ·mol-1, which proved that Ag2O and with hemp fibers formed a stable deposition. The Ag2O-hemp fibers modified electrospun polylactic acid nanofibrous composite film exhibited favorable mechanical properties. The tensile strength reached 5.23 ± 0.05 MPa and the elongation at break reached 105.56 ± 3.95 %. The obtained nanofibrous composite film has good antibacterial activity against E. coli, S. aureus, A. niger, and Penicillium, which indicated that they could effectively inhibit the growth of bacteria and fungi. The cell experiments proved that the nanofibrous composite film had good biocompatibility with a cell survival rate of 100 %. The experimental results on the fresh-keeping of red grapes showed that the PLA nanofibrous composite film modified by the Ag2O-hemp fibers could effectively prolong the spoilage time of red grapes at room temperature. Compared with the blank group, the freshness period of PLA nanofiber film modified by Ag2O-hemp fibers could be extended for more than 5 days. The hardness of 15 days (1.94 ± 0.19 × 105 Pa) was basically the same as that of 1 day (2.05 ± 0.06 × 105 Pa). The results were superior to commercially available PE preservation films. The above research results indicated that the Ag2O-hemp fibers modified PLA nanofibrous composite film had potential application prospects in the field of fruit fresh-keeping package.

Porcine epidemic diarrhea virus activates PERK-ROS axis to benefit its replication in Vero E6 cells.

Of the three branches of unfolded protein response (UPR) that were reportedly activated by porcine epidemic diarrhea virus (PEDV), PERK is recently shown to act as an upstream regulator of oxidative response of the cells. However, it remains unknown if and how PERK activation during PEDV infection would result in oxidative stress, and whether activation of PERK and its downstream molecules affect PEDV replication. Here, we demonstrate that infection with the PEDV strain YJH/2015 triggered UPR in Vero E6 cells by activating the PERK/eIF2α pathway and led to significant increase in the expression of proapoptotic protein C/EBP homologous protein (CHOP) and ER oxidoreductase 1 alpha (ERO1α). Inhibition of PERK by short hairpin RNA (shRNA) or GSK2606414 and knockdown of CHOP by small interfering RNA reduced expression of ERO1α and generation of ROS in PEDV-infected cells. Inhibition of ERO1α by shRNA or EN460 decreased PEDV-induced ROS generation. Genetic or pharmacological inhibition of each component of PERK, CHOP, ERO1α, and ROS led to significant suppression of PEDV replication. Collectively, our study provides the first evidence that PEDV manipulates endoplasmic reticulum to perturb its redox homeostasis via the PERK-CHOP-ERO1α-ROS axis in favor of its replication.