Molecular mechanisms of human coronavirus NL63 infection and replication.

Human coronavirus NL63 (HCoV-NL63) is spread globally, causing upper and lower respiratory tract infections mainly in young children. HCoV-NL63 shares a host receptor (ACE2) with severe acute respiratory syndrome coronavirus (SARS-CoV) and SARS-CoV-2 but, unlike them, HCoV-NL63 primarily develops into self-limiting mild to moderate respiratory disease. Although with different efficiency, both HCoV-NL63 and SARS-like CoVs infect ciliated respiratory cells using ACE2 as receptor for binding and cell entry. Working with SARS-like CoVs require access to BSL-3 facilities, while HCoV-NL63 research can be performed at BSL-2 laboratories. Thus, HCoV-NL63 could be used as a safer surrogate for comparative studies on receptor dynamics, infectivity and virus replication, disease mechanism, and potential therapeutic interventions against SARS-like CoVs. This prompted us to review the current knowledge on the infection mechanism and replication of HCoV-NL63. Specifically, after a brief overview on the taxonomy, genomic organization and virus structure, this review compiles the current HCoV-NL63-related research in virus entry and replication mechanism, including virus attachment, endocytosis, genome translation, and replication and transcription. Furthermore, we reviewed cumulative knowledge on the susceptibility of different cells to HCoV-NL63 infection in vitro, which is essential for successful virus isolation and propagation, and contribute to address different scientific questions from basic science to the development and assessment of diagnostic tools, and antiviral therapies. Finally, we discussed different antiviral strategies that have been explored to suppress replication of HCoV-NL63, and other related human coronaviruses, by either targeting the virus or enhancing host antiviral mechanisms.

Efficacy of ultraviolet C exposure for inactivating Senecavirus A on experimentally contaminated surfaces commonly found on swine farms.

The objective of this study was to evaluate the efficacy of ultraviolet C light (UVC) for inactivating Senecavirus A (SVA) on three different experimentally contaminated surfaces commonly found in swine farms. An experimental study under controlled conditions assessed the effect of UVC on an SVA isolate on coupons composed of three surface types: cardboard, cloth, and plastic. Each coupon was inoculated with 2 mL of SVA (107.5 TCID50/mL) and 1 mL of PBS or 1 g of feces on the top or bottom surface of the coupon and allowed to dry (90 min at 25℃). Coupons were exposed to UVC in a commercially available pass-through chamber (PTC) for 5 min or in a simulated supply entry room (SER) for 120 min. After exposure, virus isolation was attempted from each coupon and virus titers were determined in cell culture. The efficacy of UVC was determined by the reduction in virus titer for the UVC treated groups compared to their respective non-treated positive controls. UVC was effective at inactivating SVA on plastic surface free of organic material. The plastic coupons inoculated with SVA and PBS had a significantly lower virus titer (>7-log reduction) in both the PTC and SER when compared to their relative positive controls. All other groups in the PTC and SER had a 2-log reduction or less. The reduction in virus titer on the top and bottom inoculated surfaces, following exposure to UVC, were not statistically different. The data from this study provide some guidance when applying UVC for disinfection in the field.

Glucocorticoid and dietary effects on mucosal microbiota in canine inflammatory bowel disease.

The pathogenesis of canine inflammatory bowel disease (IBD) involves complex interactions between mucosal immunity and the intestinal microbiota. Glucocorticoids are commonly administered to reduce mucosal inflammation and gastrointestinal signs. The study objective was to evaluate the effects of diet and oral prednisone on the spatial distribution of mucosal bacteria in IBD dogs. Eight dogs diagnosed with IBD were treated with immunosuppressive doses of prednisone. The mucosal microbiota from endoscopic biopsies of IBD dogs and healthy controls (HC; n = 15 dogs) was evaluated by fluorescence in situ hybridization (FISH) targeting the 16S rRNA genes of total bacteria and bacterial species relevant in canine/human IBD. Apicaljunction protein (AJP) expression using immunohistochemistry investigated the effect of medical therapy on intestinal barrier integrity. All IBD dogs had a reduction in GI signs following diet and prednisone therapy compared with baseline CIBDAI scores (P < 0.05). The mucosal microbiota of HC and diseased dogs was most abundant in free and adherent mucus. Only Lactobacilli were increased (P < 0.05) in the adherent mucus of IBD dogs compared to HC. The spatial distribution of mucosal bacteria was significantly different (P < 0.05) in IBD dogs following prednisone therapy, with higher numbers of Bifidobacteria and Streptococci detected across all mucosal compartments and increased numbers of Bifidobacterium spp., Faecalibacterium spp., and Streptococcus spp. present within adherent mucus. Differences in intestinal AJPs were detected with expression of occludin increased (P < 0.05) in IBD dogs versus HC. The expressions of occludin and E-cadherin were increased but zonulin decreased (P < 0.05 for each) in IBD dogs following prednisone therapy. In conclusion, the spatial distribution of mucosal bacteria differs between IBD and HC dogs, and in response to diet and glucocorticoid administration. Medical therapy was associated with beneficial changes in microbial community structure and enhanced mucosal epithelial AJP expression.

Mortality Patterns in a Commercial Wean-To Finish Swine Production System.

Modern commercial pig production is a complex process that requires successful producers to understand and resolve factors associated with perturbations in production. One important perturbation is inventory loss due to mortality. In this study, data on 60 lots of approximately 2000 weaned pigs (n = 115,213) from one commercial production system were collected through the wean-to-finish (WTF) cycle with the objective of establishing patterns of mortality, estimating differences in profit/loss among patterns of mortality, and identifying production practices associated with mortality patterns. Information provided by the production system included the number of pigs in each lot at the time of placement (beginning inventory), weaning weight, barn dimensions, number of dead pigs (NDP) daily, capacity placed (proportion pigs actually placed versus what had been planned to be placed) and average weight sold. Analysis of NDP revealed three mortality patterns (clusters I, II, III) composed of 6, 40, and 14 lots, respectively, that differed in the temporal onset and/or level of mortality. Average daily gain (ADG) and feed conversion ratio (FCR) were calculated by growth phase for each cluster. An economic model showed profit differences among clusters due to poor biological performance by clusters I and III in the late finishing phase. Cluster II (n = 40) had fewer dead pigs and the highest profit compared to clusters I (n = 6) and III (n = 14). Area per pig (stocking density) was the only factor associated with the differences in mortality patterns. Routine monitoring and the analysis of mortality patterns for associations with production and management factors can help swine producers improve biological performance and improve profit.

Evaluation of pyramid training as a method to increase diagnostic sampling capacity during an emergency veterinary response to a swine disease outbreak.

OBJECTIVE To develop and evaluate a pyramid training method for teaching techniques for collection of diagnostic samples from swine. DESIGN Experimental trial. SAMPLE 45 veterinary students. PROCEDURES Participants went through a preinstruction assessment to determine their familiarity with the equipment needed and techniques used to collect samples of blood, nasal secretions, feces, and oral fluid from pigs. Participants were then shown a series of videos illustrating the correct equipment and techniques for collecting samples and were provided hands-on pyramid-based instruction wherein a single swine veterinarian trained 2 or 3 participants on each of the techniques and each of those participants, in turn, trained additional participants. Additional assessments were performed after the instruction was completed. RESULTS Following the instruction phase, percentages of participants able to collect adequate samples of blood, nasal secretions, feces, and oral fluid increased, as did scores on a written quiz assessing participants' ability to identify the correct equipment, positioning, and procedures for collection of samples. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that the pyramid training method may be a feasible way to rapidly increase diagnostic sampling capacity during an emergency veterinary response to a swine disease outbreak.