Immune response enhancement by dietary supplementation with Caesalpinia sappan extract in weaned pigs challenged with porcine reproductive and respiratory syndrome virus.

BACKGROUND: At present, porcine reproductive and respiratory syndrome (PRRS) caused by the PRRS virus (PRRSV) is one of the most severe epidemics impacting pig farming globally. Despite the fact that a number of studies have been conducted on potential solutions to this problem, none have proven effective. The focus of problem solving is the use of natural ingredients such as plant extracts. Popular throughout Asia, Caesalpinia sappan (CS) is a therapeutic plant that inhibits PRRSV in vitro. Therefore, this study was performed to determine the efficacy of CS extract dietary supplementation on the productive performance, antibody levels, immunological indicators, and lung pathology of PRRSV-challenged weaned pigs. A total of 32 weaned piglets (28 days old) were randomized into 4 groups and kept separately for 14 days. The treatments were organized in a 2 × 2 factorial design involving two factors: PRRSV challenge and supplementation with 1 mg/kg CS extract. The pigs in the PRRSV-challenged groups were intranasally inoculated with 2 mL of PRRSV (VR2332) containing 104 TCID50/mL, while those in the groups not challenged with PRRSV were inoculated with 2 mL of normal saline. RESULTS: In the PRRSV-challenged group (CS + PRRSV), supplementation with CS extract led to an increase in white blood cells (WBCs) on Day 7 post infection (p < 0.05) and particularly in lymphocytes on Days 7 and 14. The antibody titer was significantly greater in the CS + PRRSV group than in the PRRSV-challenged group not administered CS (PRRSV group) on Day 14 postinfection (S/P = 1.19 vs. 0.78). In addition, CS extract administration decreased the prevalence of pulmonary lesions, which were more prevalent in the PRRSV-challenged pigs that did not receive the CS extract. CONCLUSION: The findings of this study suggest that supplementation with CS extract is beneficial for increasing WBC counts, especially lymphocytes, increasing the levels of antibodies and reducing the prevalence of lung lesions in PRRSV-infected pigs.

Oral Supplementation of Houttuynia cordata Extract Reduces Viremia in PRRSV-1 Modified-Live Virus-Vaccinated Pigs in Response to the HP-PRRSV-2 Challenge.

This study evaluated the in vitro antiviral activities and the ex vivo immunomodulatory effects of Houttuynia cordata Thunb. (HC) ethanolic extracts in response to porcine reproductive and respiratory syndrome virus (PRRSV). In addition, this study evaluated the in vivo effects of oral supplementation of HC extract on immune responses to and cross-protective efficacy of PRRSV-1 modified-live virus (MLV) vaccine against the highly pathogenic (HP)-PRRSV-2 challenge. In vitro experiments demonstrated that HC extracted in either 50%, 70%, or 95% ethanol (referred to as HC50, HC70, and HC95, respectively) significantly interfered with PRRSV replication in MARC-145 cells. Ex vivo experiments revealed that all HC extracts significantly enhanced mRNA expressions of type I interferon-regulated genes, type I and II interferon (IFN), and pro- and anti-inflammatory cytokines in HP-PRRSV-2-inoculated monocyte-derived macrophages. An in vivo experiment included four groups of six pigs (4 weeks old; n = 24). Group 1 and group 2 were vaccinated with the PRRSV-1 MLV vaccine at 0 dpv (day post vaccination). Group 2 also received oral administration of HC50 extract at 0-49 dpv. Group 3 received the PRRSV-1 MLV vaccine solvent at 0 dpv, while group 4 served as strict control. Groups 1-3 were challenged intranasally with HP-PRRSV-2 at 28 dpv and immune-related and clinical parameters were monitored weekly until 49 dpv. Compared to group 1, group 2 demonstrated significantly increased IFN regulatory factor 3 mRNA expression of PRRSV-recalled peripheral blood mononuclear cells, and significantly reduced HP-PRRSV-2 viremia. No difference in PRRSV-specific antibody responses, rectal temperature, clinical scores, and average daily weight gain was detected. Our study reports the immunomodulatory and anti-PRRSV potentials of HC extract in PRRSV-1 MLV-vaccinated/HP-PRRSV-2 challenged pigs.

Oral supplementation of quercetin in PRRSV-1 modified-live virus vaccinated pigs in response to HP-PRRSV-2 challenge.

This study evaluated the immunomodulatory effect of quercetin on improving cross protection of porcine reproductive and respiratory syndrome virus-1 (PRRSV-1) modified-live virus (MLV) vaccine against highly pathogenic (HP)-PRRSV-2 challenge. Ex vivo experiments demonstrated that quercetin significantly enhanced type I interferon-regulated genes (IRGs) and type I and II interferon (IFN), and significantly decreased pro- and anti-inflammatory cytokine expressions in HP-PRRSV-inoculated monocyte-derived macrophages. In vivo experiments divided pigs (4-week-old; n = 24) into four groups of six pigs. Group 1 and group 2 were immunized with PRRSV-1 MLV vaccine at 0 dpv (day post vaccination). Group 2 also received oral administration of quercetin at 0-49 dpv. Group 3 was injected with PRRSV-1 MLV vaccine solvent at 0 dpv. Group 4 served as strict control. Group 1-3 were challenged intranasally with HP-PRRSV at 28 dpv and immune and clinical parameters were monitored weekly until 49 dpv. Group 1 demonstrated significantly reduced HP-PRRSV viremia, rectal temperature and clinical scores, and significantly improved average daily weight gain (ADWG), compared to group 3. Group 2 demonstrated significantly increased IFN regulatory factor 3, stimulator of IFN genes, IFNα, and significantly decreased transforming growth factor beta (TGFß) mRNA expressions, compared to group 1. The animals demonstrated significantly reduced HP-PRRSV viremia, but did not demonstrate any further improved PRRSV-specific antibody responses, rectal temperature, clinical scores, and ADWG as compared to group 1. Our findings suggest that quercetin up-regulates IRGs, IFNα, and down-regulates TGFß mRNA expressions which may contribute to further reducing number of viremic pigs and HP-PRRSV viremia which were conferred by PRRSV-1 MLV vaccine. Our findings also suggest that quercetin may serve as an effective oral immunomodulator for improving cell-mediated immune defense to HP-PRRSV.

The potential adjuvanticity of quaternized chitosan hydrogel based microparticles for porcine reproductive and respiratory syndrome virus inactivated vaccine.

Infectious diseases possess a big threat to the livestock industry worldwide. Currently, inactivated veterinary vaccines have attracted much attention to prevent infection due to their safer profile compared to live attenuated vaccine. However, its intrinsic poor immunogenicity demands the incorporation of an adjuvant. Mineral oil based adjuvant (Montanide™ ISA206) was usually used to potentiate the efficacy of veterinary vaccines. However, ISA206 could not induce robust cellular immune responses, which was very important in controlling virus replication and clearing the infected cells. Moreover, mineral oil would result in severe side effects. To improve both the humoral and cellular immune responses of porcine reproductive and respiratory syndrome virus (PRRSV) inactivated vaccine, we developed pH-sensitive and size-controllable quaternized chitosan hydrogel microparticles (Gel MPs) without using chemical cross linking agent. Gel MPs, ionic cross-linked with glycerophosphate (GP), were biocompatible and could efficiently adsorb the inactivated PRRSV vaccine with a loading capacity of 579.05µg/mg. After intramuscular immunization in mice, results suggested that Gel MPs elicited significantly higher cell-mediated immune responses and comparable humoral immune responses compared to ISA 206. Regarding the biocompatibility, safety and effectiveness, Gel MPs would be a promising candidate to enhance the efficacy of veterinary vaccine.

Potentiation of Taishan Pinus massoniana pollen polysaccharide on the immune response and protection elicited by a highly pathogenic porcine reproductive and respiratory syndrome virus glycoprotein 5 subunit in pigs.

Porcine reproductive and respiratory syndrome virus (PRRSV) heavily affects the global pork industry. Current available vaccine strategies have inherent drawbacks. In this work, the immune enhancement from Taishan Pinus massoniana pollen polysaccharide (TPPPS) and Freund's adjuvant on the efficacy of a PRRSV subunit vaccine were examined. Titers of specific anti-highly pathogenic PRRSV (HP-PRRSV) ELISA antibody and neutralizing antibody were significantly higher in pigs from the groups inoculated with medium- and high-dose TPPPS (mTPPPS, hTPPPS) adjuvant co-administered with a recombinant HP-PRRSV glycoprotein 5 subunit (GP5) than those from other groups (P < 0.05). Pigs inoculated with GP5 + Freund's adjuvant developed severely delayed humoral immune responses specific to GP5 within 28 days post-inoculation (dpi). The groups treated with mTPPPS and hTPPPS adjuvant exhibited the most potent immune enhancement effects on GP5 inoculation with cellular immunity developing, as shown by the level of T lymphocyte proliferation and the percentage of the CD3(+) T lymphocyte subpopulation. Although complete Freund's adjuvant elicited cell-mediated immune responses, the level of T lymphocyte proliferation in this group decreased quickly and no significant differences were observed compared with other adjuvant-alone groups at 56 dpi (P > 0.05). The ratio between CD3(+)CD4(+) and CD3(+)CD8(+) T lymphocyte subpopulations indicated the inoculums of GP5 + mTPPPS and GP5 + hTPPPS induced consistently higher CD3(+)CD4(+) T lymphocyte subpopulations than other inoculums (P < 0.05). The immune responses caused by complete Freund's adjuvant were mainly mediated by CD3(+)CD8(+) T lymphocyte subpopulation (cytotoxic T lymphocytes) in the early stage of inoculation and had no significant difference compared with other adjuvant-alone groups after 28 dpi (P > 0.05). The low-dose TPPPS (lTPPPS) adjuvant also exhibited enhancement effects on humoral immune and T lymphocyte proliferation responses but these were significantly lower than the mTPPPS and hTPPPS doses (P < 0.05). Pigs challenged with HP-PRRSV from the GP5 + mTPPPS, GP5 + hTPPPS, and GP5 + Freund's adjuvant groups showed lower viremia, fewer clinical signs, and fewer pathological lung lesions compared with the groups of GP5-alone and GP5 + lTPPPS (P < 0.05). There were significant differences between the GP5-alone and GP5 + lTPPPS groups in detection indexes after viral challenge (P < 0.05). In conclusion, moderate doses of TPPPS as an adjuvant with GP5 show promise as a candidate for a HP-PRRSV subunit vaccine to efficiently prevent and control HP-PRRSV.

Dietary conjugated linoleic acid and its effect on immune response in pigs infected with the porcine reproductive and respiratory syndrome virus.

The aim of this study was to evaluate the effects of dietary conjugated linoleic acid (CLA) on immune response in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV). A total of 18 pigs 4 weeks of age were allocated to 3 treatments, 6 per treatment: 0% CLA, 1% CLA, and 2% CLA. Serum IL-1ß, IL-6 and TNF-α; lymphocyte proliferation; and IL-2, IFN-γ, IL-10, IL-4 and IL-12 in PBMCs were evaluated. NF-κB, COX2, iNOS and PPAR-γ mRNA were also evaluated. No differences were observed among treatment groups in most of the in vivo cytokine profiles; only TNF-α production was increased in infected pigs in the CLA-supplemented groups. The cytokine profile in vitro was not affected by CLA supplementation. CLA decreased the proliferation of PBMCs stimulated with PRRSVs. Inflammation mediators and PPAR-γ were not affected by CLA in infected pigs. CLA did not improve the immune response of PRRSV infected pigs.

Effects of dietary threonine and tryptophan supplementation on growing pigs induced by porcine respiratory and reproductive syndrome vaccination.

A total of 32 growing pigs were used in a 2 × 2 factorial arrangement of treatments with two different diets (conventional [CON] diet vs. threonine [Thr]- and tryptophan [Trp]-rich [TTR] diet) and two immunological challenge regimens (porcine respiratory and reproductive syndrome [PRRS] vaccine vs. phosphate buffer solution [PBS]) to study the hypothesis that dietary supplementation with Trp and Thr would benefit for growing pigs vaccinated with PRRS vaccine. After feeding the experimental diets for 21 d, the pigs were intramuscularly vaccinated with PRRS or PBS. Performance data were recorded over a period of 10 weeks and are presented for the pre-challenge period (3 weeks) and the challenge period (7 weeks, where on day 1, pigs were immunologically challenged). During the pre-challenge period, the growth performance was not different between dietary treatments. PRRS vaccination resulted in increased rectal temperature and decreased feed intake and growth rate (p < 0.05). In PRRS-vaccinated pigs, diet TTR enhanced the feed intake, especially during the first 2 weeks after the PRRS vaccination compared with diet CON (p < 0.05). PRRS vaccination also resulted in increased plasma concentration of urea nitrogen, essential and non-essential amino acids (p < 0.05) and porcine reproductive and respiratory syndrome virus specific antibodies (p < 0.05), but decreased concentration of immunoproteins including alpha-1-acylglycoprotein and immunoglobulin G (p < 0.05). The alleviation of the PRRS vaccination induced decrease in feed intake and growth rate by Thr and Trp supplementation, indicating that the PRRS-vaccinated pigs had a higher Thr and Trp requirement than non-vaccinated pigs.

Elevated dietary zinc oxide levels do not have a substantial effect on porcine reproductive and respiratory syndrome virus (PPRSV) vaccination and infection.

BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important infectious agents for the swine industry worldwide. Zinc (Zn) salts, which are widely used as a dietary supplement in swine nutrition, have shown antiviral effects in vitro as well as in vivo. The purpose of this study was to determine the influence of dietary zinc oxide supplementation on vaccination and challenge infection with PRRSV. FINDINGS: The clinical course of PRRS and the success of vaccination with an experimental inactivated vaccine were compared between animals receiving a conventional diet (50 ppm Zn, control group) and diets supplemented with Zn oxide (ZnO) at final Zn concentrations of 150 or 2,500 ppm. Pigs receiving higher dietary Zn levels showed a tendency towards higher neutralizing antibody levels after infection, while dietary Zn levels did not substantially influence the number of antiviral IFN-gamma secreting cells (IFN-gamma-SC) or percentages of blood immune cell subsets after infection. Finally, feeding higher dietary Zn levels reduced neither clinical symptoms nor viral loads. CONCLUSIONS: Our results suggest that higher levels of dietary ZnO do not have the potential to stimulate or modulate systemic immune responses after vaccination and heterologous PRRSV infection to an extent that could improve the clinical and virological outcome.

Vitamin E supplementation does not mitigate the acute morbidity effects of porcine reproductive and respiratory syndrome virus in nursery pigs.

The objective of this study was to determine whether feeding a vitamin E-rich diet would benefit nursery pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV). Sixty-four pigs were subjected to one of four treatment combinations (2 x 2 factorial) of dietary vitamin E (adequate or excess) and PRRSV (medium or inoculation with VR-2385 isolate P-129). Pigs were fed experimental diets during a 3-wk period before inoculation as well as during a 12-d period after inoculation. Growth performance was determined throughout the study, and lipid peroxidation in liver, glutathione peroxidase (GPX) activity in serum, circulating white blood cells, and serum interleukin-1beta (IL-1beta) and interferon-gamma (IFN-gamma) were determined in samples collected from pigs killed 4 or 12 d after inoculation. Infection by PRRSV (P < 0.001) induced a marked decrease in both ADFI and ADG, but neither the main effect of diet nor the diet x PRRSV interaction was significant. Neither diet nor PRRSV affected feed efficiency. At 12 d after inoculation, lipid peroxidation in liver and GPX activity in serum were lower in pigs fed excess vitamin E than in those fed adequate vitamin E (P < 0.01), suggesting that the diet high in vitamin E bolstered the antioxidant status of the pigs. However, PRRSV did not affect lipid peroxidation in liver or serum GPX activity, and the diet x PRRSV interaction was not significant. White blood cell counts were decreased and IFN-gamma, and IL-1beta were increased (P < 0.05) 4 and 12 d after inoculation in PRRSV-infected pigs, but neither diet nor the diet x PRRSV interaction was significant. Collectively, these results indicate that increasing antioxidant defenses by feeding high levels of vitamin E did not ameliorate the effects of PRRSV on decreased growth, leukopenia, and increased serum IL-1beta and IFN-gamma. Thus, feeding nursery pigs a diet high in vitamin E may not be useful for mitigating the acute morbidity effects of PRRSV infection.

Effect of dietary Echinacea purpurea on viremia and performance in porcine reproductive and respiratory syndrome virus-infected nursery pigs.

The effect of dietary Echinacea purpurea on performance, viremia, and ontogeny of the humoral antibody response against porcine reproductive and respiratory syndrome virus (PRRSV) infection was evaluated in weaned pigs. In three replicates, 120 weaned pigs (25 +/- 1 d of age; 8.46 +/- 0.48 kg of BW) from a PRRSV-naive herd were allotted randomly to one of eight pens (diets) in two separate rooms (four pens/room), with each pen containing five pigs. Pigs began one of four dietary treatments (as-fed basis) 1 wk before inoculation with PRRSV: 1) basal diet composed of corn, soybean meal, whey, and essential vitamins and minerals; 2) basal diet plus carbadox (0.055 g/kg of diet; as-fed basis); 3) basal diet plus Echinacea 2% (2% of the total diet); 4) basal diet plus Echinacea 4% (4% of the total diet). The diets were formulated to be isocaloric and isolysinic. Echinacea purpurea was purchased in powder form and determined by chemical analysis to contain 1.35% cichoric acid (as-fed basis). Seven days after starting the diets, all pigs in one room were intranasally inoculated with PRRSV isolate ATCC VR-2332 at a concentration of 10(4) tissue culture infectious dose50/mL. To monitor the effects of Echinacea and PRRSV challenge, BW and blood samples were obtained from all pigs at 7-d intervals. Serum samples were analyzed for the presence of PRRSV and PRRSV-specific antibodies. All challenged pigs became infected with PRRSV, and all unchallenged pigs remained free of infection. No differences (P > 0.10) in ADG, ADFI, or gain:feed (G:F) were observed in PRRSV-challenged compared with unchallenged animals. For PRRSV-challenged animals receiving diets supplemented with Echinacea at 2 or 4%, no differences (P > 0.10) were observed in ADG, ADFI, or G:F ratio. Among PRRSV-challenged pigs, dietary Echinacea did not affect (P > 0.10) the rate or level of the ELISA-detectable antibody response from d 7 to 42 or the level and duration of PRRSV in serum. For PRRSV-unchallenged animals receiving diets supplemented with Echinacea at 2 or 4%, no differences (P > 0.10) were observed in ADG, ADFI, and G:F ratio. Under the conditions of this study, dietary Echinacea did not enhance growth, exhibit antiviral effects to PRRSV, or show any evidence of immune enhancing properties.