Antiviral activity against porcine epidemic diarrhea virus of Pogostemon cablin polysaccharide.

ETHNOPHARMACOLOGICAL RELEVANCE: The dry overground parts of Pogostemon cablin (Blanco) Benth. is widely used in China as a traditional Chinese medicine for the treatment of diarrhea, vomiting, nausea and fever. Polysaccharide is an important component of Pogostemon cablin (Blanco) Benth. but has not been studied. Pogostemon cablin (Blanco) Benth. is used to treat porcine epidemic diarrhea. But it is not known whether Pogostemon cablin polysaccharides (PCPs) has the antiviral activities against porcine epidemic diarrhea virus (PEDV). AIM OF THE STUDY: The purpose of present study is to investigate the structural characterization and the anti-PEDV activities of PCPs. MATERIALS AND METHODS: PCPs were prepared by water extraction and alcohol precipitation method and purified with DEAE-52 cellulose column and Sephadex G-100 column. Then, the structural characterization of the polysaccharides including the infrared spectrum, molecular weight and monosaccharide composition were analyzed. Afterwards, the antiviral effect of PCPs against PEDV on IPEC-J2 cells was studied by MTT method and real-time PCR method. Additionally, the effects of PCPs on PEDV adsorption, penetration and replication were analyzed by real-time PCR method. Furthermore, we also investigate whether the anti-oxidative effects of PCPs were important to the anti-PEDV activities. RESULTS: Four polysaccharides were obtained and named as PCP1.1 (31.3 kDa), PCP1.2 (3.5 kDa), PCP2.1 (9.1 kDa) and PCP2.2 (8.3 kDa). PCP1.1, PCP1.2 and PCP2.1 were composed of fucose, arabinose, galactose, glucose, mannose, galacturonic acid and glucuronic acid; and PCP2.2 was composed of arabinose, galactose, glucose, galacturonic acid and glucuronic acid. All PCPs showed anti-PEDV activities. PCP1.1 and PCP1.2 inhibited PEDV replication, while PCP2.1 and PCP2.2 inhibited PEDV penetration and replication. All PCPs showed anti-oxidative effects, which were important to the anti-PEDV activities. CONCLUSIONS: The treatment effect of Pogostemon cablin (Blanco) Benth. on porcine epidemic diarrhea might be related to the anti-PEDV effect of PCPs. Furthermore, the anti-oxidative effects of PCPs play important roles in their antiviral activities against PEDV.

Passive immunity to porcine epidemic diarrhea virus following immunization of pregnant gilts with a recombinant orf virus vector expressing the spike protein.

Passive immunity is critical for protection of neonatal piglets against porcine epidemic diarrhea virus (PEDV). Here, we investigated the immunogenicity of an orf virus (ORFV) vector expressing the full-length spike (S) protein of PEDV (ORFV-PEDV-S) in pregnant gilts and its ability to confer passive immunity and protection in piglets. Three doses of ORFV-PEDV-S were given to two groups of PEDV-negative pregnant gilts, with the last dose being administered two weeks prior to farrowing. One of the two groups immunized with the ORFV-PEDV-S recombinant virus was also exposed to live PEDV orally on day 31 post-immunization (pi). Antibody responses were assessed in serum, colostrum and milk of immunized gilts, and passive transfer of antibodies was evaluated in piglet sera. The protective efficacy of ORFV-PEDV-S was evaluated after challenge of the piglets with PEDV. PEDV-specific IgG, IgA and neutralizing antibody (NA) responses were detected in ORFV-PEDV-S-immunized and ORFV-PEDV-S-immunized/PEDV-exposed gilts. PEDV NA, IgG and IgA were detected in the serum of piglets born to immunized gilts, demonstrating the transfer of antibodies through colostrum and milk. Piglets born to immunized gilts showed reduced morbidity and a marked reduction in mortality after PEDV challenge in comparison to control piglets. Piglets born to gilts that received ORFV-PEDV-S and were exposed to live PEDV showed stronger NA responses and lower clinical scores when compared to piglets born to gilts immunized with ORFV-PEDV-S alone. These results demonstrate the potential of ORFV as a vaccine delivery platform capable of eliciting passive immunity against PEDV.

Pig epidemic diarrhoea virus S gene variant with a large deletion non-lethal to colostrum-deprived newborn piglets.

We previously identified a third porcine epidemic diarrhoea virus (PEDV) S variant with a large deletion of 582 nucleotides in the 5' terminal region of the S gene, in addition to the North American type and the S INDELs type. To investigate the pathogenicity of this variant, TTR-2/JPN/2014, we performed experimental infection using colostrum-deprived piglets and compared the results with those from the North American type PEDV, OKN-1/JPN/2013. Fifteen newborn piglets were divided into two groups of 7-8 piglets each and inoculated orally with one of PEDV isolates maintained at the eighth passage in Vero cell culture. Although all PEDV-inoculated piglets showed acute watery diarrhoea, lethality clearly differed between both PEDV-inoculated groups. Moreover, there were differences in virus distribution and lesions on the intestines between the two PEDV-inoculated groups. Therefore, our data suggest that the OKN-1/JPN/2013 PEDV isolate is virulent, whereas the TTR-2/JPN/2014 PEDV isolate is avirulent.

Characterization of Porcine Epidemic Diarrhea Virus Isolate US/Iowa/18984/2013 Infection in 1-Day-Old Cesarean-Derived Colostrum-Deprived Piglets.

Porcine epidemic diarrhea virus (PEDV) was first recognized in North America in April 2013 and has since caused devastating disease. The objective of this study was to characterize disease and viral detection associated with an original North American PEDV isolate inoculated in neonatal piglets. Thirty-six 1-day-old cesarean-derived and colostrum-deprived piglets were randomly assigned to the control (n = 16) or challenged group (n = 20); the latter were orogastrically inoculated with 1 ml of US/Iowa/18984/2013 PEDV isolate titered at 1 × 10(3) plaque-forming units per milliliter. Rectal swabs were collected from all piglets prior to inoculation and every 12 hours postinoculation (hpi) thereafter, with 4 control and 5 challenged piglets euthanized at 12, 24, 48, and 72 hpi. One piglet had a positive real-time quantitative polymerase chain reaction test on rectal swab at 12 hpi, and all remaining piglets were positive thereafter, with highest viral quantities detected at 24 and 36 hpi. Diarrhea was evident in 30% and 100% of challenged piglets at 18 and 24 hpi, respectively. Viral antigen was detected in enterocytes by immunohistochemistry in the duodenum and ileum of piglets euthanized at 12 hpi and was apparent throughout the small intestine of all piglets thereafter, with villus height:crypt depth ratios consistently below 4:1. Viremia was confirmed in 18 of 20 pigs at euthanasia. Clinical disease was severe and developed rapidly following infection with an original North American PEDV isolate, with lesions, viremia, and antigen detection possible by 12 hpi.

Antioxidative and antiviral properties of flowering cherry fruits (Prunus serrulata L. var. spontanea).

The phenolic compounds of many fruits have been known to be efficient cellular protective antioxidants. In this study, antioxidative and antiviral properties of flowering cherry cultivars (Prunus yedoensis, Prunus sargentii, Prunus lannesiana, and Prunus cerasus) in Korea were investigated. The antioxidant property was assayed for specific activities including 2,2-diphenyl-1-picrylhydrazyl (DPPH) hydroxy radical scavenging activity, reducing power capacity, and superoxide dismutase (SOD) like activity. In addition, antiviral activity was determined by inhibition studies on the infection cycle of porcine epidemic diarrhea virus (PEDV), measured as minimum concentration of cherry extracts that inhibited 50% of cytopathic effect (CPE) on PEDV. Our results show that the four varieties of cherries contain substantially high antioxidants and antiviral activities. In particular, P. cerasus contains higher antioxidants and antiviral activities as well as polyphenolic content than other varieties. Our data indicate that Korean native cherry cultivars could be beneficial supplements of dietary antioxidants and natural antiviral agents.