scholarly journals Prevalence and factors associated to contamination from porcine epidemic diarrhea virus during the transportation of pigs to slaughterhouses in Colombia

2020 ◽  
Author(s):  
Maria del Pilar Pineda Ortiz ◽  
Johanna Paola Corrales Morales ◽  
Gilma Hernández Herrera ◽  
Carlos Enrique Trillos Pena ◽  
Diana Corina Zambrano Moreno
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Animal Feed ◽  
The United States ◽  
The Body ◽  
Swine Industry ◽  
Pressurized Water ◽  
Diarrhea Virus ◽  
Factors Associated ◽  
Porcine Epidemic Diarrhea ◽  
The Impact

Abstract Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea, dehydration, and high mortality in newborn piglets and has caused high economic impact in the swine industry in the United States and Asia. Until March 2014, PED was an exotic disease in Colombia. This study was carried out at the beginning of the spread of PEDV in the country, and its main objective was to determine the prevalence and factors associated to the contamination of PEDV in the transportation of pigs to the slaughterhouses in Colombia through environmental samples analyzed by RT-PCR. 518 pig trucks in the 32 main slaughterhouses, were sampled and the drivers of the trucks fulfilled a questionnaire. The prevalence of PEDV at the entrance of the slaughterhouses was 71.8% (CI 95%: 70.8-72.8) versus 70.5% (CI 95%: 69.5-71.5) at the slaughterhouse exit, and there was no evidence of significant differences between both rates. (McNemar value p: 0.375). The factors that increased the possibility of truck contamination were: vehicles that visit national slaughterhouses (OR 15.9 95% CI: 4.9–51.85) and that visit national –export type (OR 9.0 95% CI: 2.20–36.91), trucks with mobility in area of ​​greatest slaughter (OR 9.05 95% CI: 2.9-27.63), the non-exclusive use of vehicles to transport pigs (OR 3.75 95% CI: 1.55-9.08) and visit animal feed mills (OR 13.5 95% CI: 4.1– 44.12). The factors identified that reduce the possibility of contamination were cleanliness of the body truck (OR 0.089 95% CI: 0.03-0.23) and the cabin (OR 0.16 95%CI: 0.08-0.31), use of disinfectant (OR 0.32 95% CI:0.16-0.62), pressurized water (OR 0.38 95% CI:0.15-0.95), and back pump (OR 0.17 95% CI:0.08-0.35) and the exclusive use of the vehicle for pig transportation (OR 0.36 95% CI: 0.19-0.70). The results showed that the high grade of mobilization of the trucks to points of concentration, failures in biosecurity and virus survival characteristics, enabled the spread of PEDV, turning it into an endemic disease in Colombia; however, the measures implemented by the farmers mitigated the impact of the disease. We evidenced the need of standardizing and regulating biosecurity protocols for slaughterhouses, pig transportation and farms.

Prevalence and factors associated with the contamination of porcine epidemic diarrhea virus during pig transport to slaughterhouses in Colombia

2019 ◽  
Author(s):  
Maria del Pilar Pineda Ortiz ◽  
Johanna Paola Corrales Morales ◽  
Gilma Hernández Herrera ◽  
Carlos Enrique Trillos Pena ◽  
Diana Corina Zambrano Moreno
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Animal Feed ◽  
Epidemiological Data ◽  
Cross Sectional Study ◽  
Morbidity Rate ◽  
Cross Sectional ◽  
Diarrhea Virus ◽  
Factors Associated ◽  
Proportional Allocation ◽  
Porcine Epidemic Diarrhea

Abstract Abstract: Background: Porcine epidemic diarrhea (PED) is an infectious disease that mainly affects neonatal piglets with a morbidity rate of 80%–100% and a mortality rate of 50%–90%. Prior to March 2014, PED was an exotic disease in Colombia with no historical epidemiological data and this study was conducted at the beginning of the spread of the virus in the country. The aim of the present study was to determine the prevalence and factors associated with porcine epidemic diarrhea virus (PEDV) contamination in pig transportation to slaughterhouses in Colombia through a cross-sectional study involving a sample of 518 pig trucks visiting the 32 main slaughterhouses authorized by the regulatory agency of the govern, the INVIMA. Samples were obtained using proportional allocation stratified sampling of pig trucks entering and leaving slaughterhouses for the diagnosis of PEDV through RT-PCR, as well as conducting a survey of the drivers of the sampled trucks. The sampling was performed from June to November 2014, during the first outbreak. Results: The prevalence of PEDV at the time of entry of the plant was 71.8% (CI 95%: 70.8–72.8) versus 70.5% (CI 95%: 69.5–71.5) on exit (p=0.375). Associated factors found to increase the possibility of contamination included: the type of slaughterhouse either national (OR 15.9, CI 95%: 4.9–51.85) or national–exportation (OR 9.0, CI 95%: 2.20–36.91), the zone of highest slaughter (OR 9.05, CI 95%: 2.9–27.63), the non-exclusive use of vehicles for the transport of pigs (OR 3.75, CI 95%: 1.55–9.08) and visiting animal feed plants (OR 13.5, CI 95%: 4.1–44.12). Factors identified to reduce the possibility of contamination included: the cleaning of vehicles, the use of disinfectants on the vehicle and the exclusive of the vehicle for pig transportation. Conclusions: The results showed that the high degree of truck mobilization, with poor biosafety compliance and dissemination characteristics of the virus, facilitated the spread of PEDV throughout the national territory. These factors contributed to establishing the disease as an endemic problem in Colombia. Keywords: coronavirus, epidemiology, PEDV, prevalence, swine.

scholarly journals Effect of pelleting on survival of porcine epidemic diarrhea virus–contaminated feed

2017 ◽  
Vol 95 (3) ◽  
pp. 1170-1178 ◽  
Author(s):  
R. A. Cochrane ◽  
L. L. Schumacher ◽  
S. S. Dritz ◽  
J. C. Woodworth ◽  
A. R. Huss ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Thermal Processing ◽  
Porcine Epidemic Diarrhea Virus ◽  
High Dose ◽  
Heat Sensitivity ◽  
Swine Industry ◽  
Diarrhea Virus ◽  
Reverse Transcription Pcr ◽  
Porcine Epidemic Diarrhea ◽  
The Impact

Abstract Porcine epidemic diarrhea virus (PEDV) is a heat-sensitive virus that has devastated the U.S. swine industry. Because of its heat sensitivity, we hypothesized that a steam conditioner and pellet mill mimicking traditional commercial thermal processing may mitigate PEDV infectivity. Pelleting, a common feed processing method, includes the use of steam and shear forces, resulting in increased temperature of the processed feed. Two thermal processing experiments were designed to determine if different pellet mill conditioner retention times and temperatures would impact PEDV quantity and infectivity by analysis of quantitative reverse transcription PCR and bioassay. In Exp. 1, a 3 · 3 · 2 factorial design was used with 3 pelleting temperatures (68.3, 79.4, and 90.6°C), 3 conditioning times (45, 90, or 180 s), and 2 doses of viral inoculation (low, 1 · 102 tissue culture infectious dose50 (the concentration used to see cytopathic effect in 50% of the cells)/g, or high, 1 · 104 tissue culture infectious dose50/g). Noninoculated and PEDV-inoculated unprocessed mash were used as controls. The low-dose PEDV–infected mash had 6.8 ± 1.8 cycle threshold (Ct) greater (P < 0.05) PEDV than the high-dose mash. Regardless of time or temperature, pelleting reduced (P < 0.05) the quantity of detectable viral PEDV RNA compared with the PEDV-inoculated unprocessed mash. Fecal swabs from pigs inoculated with the PEDV-positive unprocessed mash, regardless of dose, were clinically PEDV positive from 2 to 7 d (end of the trial) after inoculation. However, if either PEDV dose of inoculated feed was pelleted at any of the 9 tested conditioning time · temperature combinations, no PEDV RNA was detected in fecal swabs or cecum content. Based on Exp. 1 results, a second experiment was developed to determine the impact of lower processing temperatures on PEDV quantity and infectivity. In Exp. 2, PEDV-inoculated feed was pelleted at 1 of 5 conditioning temperatures (37.8, 46.1, 54.4, 62.8, and 71.1°C) for 30 s. The 5 increasing processing temperatures led to feed with respective mean Ct values of 32.5, 34.6, 37.0, 36.5, and 36.7, respectively. All samples had detectable PEDV RNA. However, infectivity was detected by bioassay only in pigs from the 37.8 and 46.1°C conditioning temperatures. Experiment 2 results suggest conditioning and pelleting temperatures above 54.4°C could be effective in reducing the quantity and infectivity of PEDV in swine feed. However, additional research is needed to prevent subsequent recontamination after pelleting as it is a point-in-time mitigation step.

scholarly journals Gene Variations in Cis-Acting Elements between the Taiwan and Prototype Strains of Porcine Epidemic Diarrhea Virus Alter Viral Gene Expression

Genes ◽  
2018 ◽  
Vol 9 (12) ◽  
pp. 591
Author(s):  
Tsung-Lin Tsai ◽  
Chen-Chang Su ◽  
Ching-Chi Hsieh ◽  
Chao-Nan Lin ◽  
Hui-Wen Chang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Porcine Epidemic Diarrhea Virus ◽  
The Body ◽  
Economic Losses ◽  
Viral Gene ◽  
Sequence Motif ◽  
Cis Acting ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea ◽  
In Cis

In 2013, the outbreak of porcine epidemic diarrhea (PED) in Taiwan caused serious economic losses. In this study, we examined whether the variations of the cis-acting elements between the porcine epidemic diarrhea virus (PEDV) Taiwan (TW) strain and the prototype strain CV777 alter gene expression. For this aim, we analyzed the variations of the cis-acting elements in the 5’ and 3’ untranslated regions (UTRs) between the PEDV TW, CV777, and other reference strains. We also determined the previously unidentified transcription regulatory sequence (TRS), a sequence motif required for coronavirus transcription, and found that a nucleotide deletion in the TW strain, in comparison with CV777 strain, immediately downstream of the leader core sequence alters the identity between the leader TRS and the body TRS. Functional analyses using coronavirus defective interfering (DI) RNA revealed that such variations in cis-acting elements for the TW strain compared with the CV777 strain have an influence on the efficiency of gene expression. The current data show for the first time the evolution of PEDV in terms of cis-acting elements and their effects on gene expression, and thus may contribute to our understanding of recent PED outbreaks worldwide.

scholarly journals Origin, Evolution, and Genotyping of Emergent Porcine Epidemic Diarrhea Virus Strains in the United States

mBio ◽  
2013 ◽  
Vol 4 (5) ◽  
Author(s):  
Yao-Wei Huang ◽  
Allan W. Dickerman ◽  
Pablo Piñeyro ◽  
Long Li ◽  
Li Fang ◽  
...  
Keyword(s):  
United States ◽  
Porcine Epidemic Diarrhea Virus ◽  
The United States ◽  
Genomic Sequences ◽  
Evolutionary Divergence ◽  
Origin And Evolution ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea ◽  
And Control ◽  
Complete Genomic

ABSTRACT Coronaviruses are known to infect humans and other animals and cause respiratory and gastrointestinal diseases. Here we report the emergence of porcine epidemic diarrhea virus (PEDV) in the United States and determination of its origin, evolution, and genotypes based on temporal and geographical evidence. Histological lesions in small intestine sections of affected pigs and the complete genomic sequences of three emergent strains of PEDV isolated from outbreaks in Minnesota and Iowa were characterized. Genetic and phylogenetic analyses of the three U.S. strains revealed a close relationship with Chinese PEDV strains and their likely Chinese origin. The U.S. PEDV strains underwent evolutionary divergence, which can be classified into two sublineages. The three emergent U.S. strains are most closely related to a strain isolated in 2012 from Anhui Province in China, which might be the result of multiple recombination events between different genetic lineages or sublineages of PEDV. Molecular clock analysis of the divergent time based on the complete genomic sequences is consistent with the actual time difference, approximately 2 to 3 years, of the PED outbreaks between China (December 2010) and the United States (May 2013). The finding that the emergent U.S. PEDV strains share unique genetic features at the 5′-untranslated region with a bat coronavirus provided further support of the evolutionary origin of PEDV from bats and potential cross-species transmission. The data from this study have important implications for understanding the ongoing PEDV outbreaks in the United States and will guide future efforts to develop effective preventive and control measures against PEDV. IMPORTANCE The sudden emergence of porcine epidemic diarrhea virus (PEDV), a coronavirus, for the first time in the United States causes significant economic and public health concerns. Since its recognition in May 2013, PEDV has rapidly spread across the United States, resulting in high mortality in piglets in more than 17 States now. The ongoing outbreaks of Middle East respiratory syndrome coronavirus in humans from countries in or near the Arabian Peninsula and the historical deadly nature of the 2002 outbreaks of severe acute respiratory syndrome coronavirus create further anxiety over the emergence of PEDV in the United States due to the lack of scientific information about the origin and evolution of this emerging coronavirus. Here we report the detailed genetic characterization, origin, and evolution of emergent PEDV strains in the United States. The results provide much needed information to devise effective preventive and control strategies against PEDV in the United States.

scholarly journals The trypsin-enhanced infection of porcine epidemic diarrhea virus is determined by the S2 subunit of the spike glycoprotein

2021 ◽  
Author(s):  
Yubei Tan ◽  
Limeng Sun ◽  
Gang Wang ◽  
Yuejun Shi ◽  
Wanyu Dong ◽  
...  
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Cleavage Site ◽  
Vaccine Development ◽  
Syncytium Formation ◽  
Reverse Genetic System ◽  
Site Mutation ◽  
Swine Industry ◽  
S Protein ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV) is an enteric pathogen in the swine industry, causing high mortality in neonatal piglets. Efficient PEDV infection usually relies on the presence of trypsin, yet the mechanism of trypsin dependency is ambiguous. Here, we identified two PEDV strains, trypsin-enhanced YN200 and trypsin-independent DR13, in which the spike (S) protein of YN200 exhibits a stronger ability to induce syncytium formation and cleaved by trypsin than that of DR13. Using a full-length infectious YN200 cDNA clone, we confirmed that the S protein is a trypsin dependency determinant by comparison of rYN200 and rYN200-SDR13. To explore the trypsin-associated sites of the YN200 S protein, we then constructed a series of mutations adjacent to the fusion peptide. The results show that the putative S2’ cleavage site (R892G) is not the determinant for virus trypsin dependency. Hence, we generated viruses carrying chimeric S proteins: the S1 subunit, S2 subunit, and S2720∼892 aa domain (NS2’) were individually replaced by the corresponding DR13 sequences. Intriguingly, only the S2 substitution, not the S1 or NS2’ substitutions, provides trypsin-independent growth of YN200. Additionally, the NS2’ recombinant virus significantly abrogated effective infection, indicating a vital role for NS2’ in viral entry. These findings suggest that the trypsin dependency of PEDV is mainly controlled by mutations in the S2 subunit rather than directly trypsin cleavage site. Importance With the emergence of new variants, PEDV remains a major problem in the global swine industry. Efficient PEDV infection usually requires trypsin, while the mechanism of trypsin dependency is complex. Here, we used two PEDV strains, trypsin-enhanced YN200 and trypsin-independent DR13, and results showed that the S protein determined PEDV trypsin dependency by using a reverse genetic system of YN200. The S2 subunit was verified as the main portion of PEDV trypsin dependency, though the putative S2’ site mutation cannot render trypsin-independent growth of YN200. Finally, these results provide some different insight to the PEDV trypsin dependency and might inspire vaccine development.

scholarly journals Single-Particle Tracking Porcine Epidemic Diarrhea Virus Moving along Microtubules in Living Cells

Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 124
Author(s):  
Yangyang Li ◽  
Wei Hou ◽  
Jian Wang ◽  
Fei Liu
Keyword(s):  
Particle Tracking ◽  
Single Particle ◽  
Porcine Epidemic Diarrhea Virus ◽  
Living Cells ◽  
Single Particle Tracking ◽  
Microtubule Organizing Center ◽  
Swine Industry ◽  
Diarrhea Virus ◽  
Organizing Center ◽  
Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV), a member of the genus Alphacoronavirus, has caused severe damage to the swine industry. Although viruses are believed to hijack the microtubule-based transport system, the exact manner of PEDV moving along microtubules has not been fully characterized. In this study, PEDV was labeled with quantum dots which have great brightness and photostability. By using quantum dot-labeled PEDV and single-particle tracking, we were able to systematically dissect the dynamic behaviors of PEDV moving along the microtubules in living cells. We found that PEDVs maintained a restricted motion mode with a relatively stable speed in the cell membrane region while displaying a slow–fast–slow velocity pattern with different motion modes in the cell cytoplasm region and near the microtubule-organizing center. The return movements of small amounts of PEDVs were also observed in living cells. Collectively, our work is crucial for understanding the movement of PEDV in living cells; the proposed work also provides important references for further analysis and studies of the infection mechanism of PEDV.

scholarly journals Molecular Characterization of Porcine Epidemic Diarrhea Virus and Its New Genetic Classification Based on the Nucleocapsid Gene

Viruses ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 790
Author(s):  
Sung-Jae Kim ◽  
Van-Giap Nguyen ◽  
Thi-My-Le Huynh ◽  
Yong-Ho Park ◽  
Bong-Kyun Park ◽  
...  
Keyword(s):  
B Cell ◽  
Porcine Epidemic Diarrhea Virus ◽  
Cell Epitope ◽  
N Gene ◽  
Swine Industry ◽  
Genetic Classification ◽  
Nucleocapsid Gene ◽  
Diarrhea Virus ◽  
B Cell Epitope ◽  
Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV) causes continuous, significant damage to the swine industry worldwide. By RT-PCR-based methods, this study demonstrated the ongoing presence of PEDV in pigs of all ages in Korea at the average detection rate of 9.92%. By the application of Bayesian phylogenetic analysis, it was found that the nucleocapsid (N) gene of PEDV could evolve at similar rates to the spike (S) gene at the order of 10−4 substitutions/site/year. Based on branching patterns of PEDV strains, three main N gene-base genogroups (N1, N2, and N3) and two sub-genogroups (N3a, N3b) were proposed in this study. By analyzing the antigenic index, possible antigenic differences also emerged in both the spike and nucleocapsid proteins between the three genogroups. The antigenic indexes of genogroup N3 strains were significantly lower compared with those of genogroups N1 and N2 strains in the B-cell epitope of the nucleocapsid protein. Similarly, significantly lower antigenic indexes in some parts of the B-cell epitope sequences of the spike protein (COE, S1D, and 2C10) were also identified. PEDV mutants derived from genetic mutations of the S and N genes may cause severe damage to swine farms by evading established host immunities.

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