Serum IgA antibody level against porcine epidemic diarrhea virus is a potential pre-evaluation indicator of immunization effects in sows during parturition under field conditions.

BACKGROUND: Porcine Epidemic Diarrhea (PED) is a highly contagious disease caused by Porcine Epidemic Diarrhea Virus (PEDV), resulting in a mortality rate of suckling piglets as high as 100%. Vaccination is the primary strategy for controlling PEDV infection, however, there is currently a lack of reliable methods for assessing the efficacy of vaccination. This study aimed to analyze serum and colostrum samples from 75 parturient sows with a specific vaccination strategy to measure levels of IgG, IgA, and neutralizing antibodies (nAbs) against PEDV, and to investigate the correlation between serum and colostrum antibody levels, as well as to identify potential biomarkers that can be used to evaluate immunization effects under field conditions. RESULTS: The findings of correlation analysis between antibody levels of IgA, IgG, and nAbs in serum or colostrum samples revealed that IgG demonstrated the most robust correlation with nAbs exhibiting a correlation coefficient of 0.64 in serum samples. Conversely, IgA exhibited the highest correlation with nAbs, with a correlation coefficient of 0.47 in colostrum samples. Additionally, the correlation analysis of antibody levels between serum and colostrum samples indicated that serum IgA displayed the strongest correlation with colostrum IgA, with a coefficient of 0.63, indicating that serum IgA may serve as a viable alternative indicator for evaluating IgA levels in colostrum samples. To further evaluate the suitability of serum IgA as a substitute marker for colostrum IgA, levels of IgA antibodies in serum samples from sows were examined both pre- and post-parturition. The findings indicated that serum IgA levels were initially low prior to the initial immunization, experienced a notable rise 21 days after immunization, and maintained a significant elevation compared to pre-immunization levels from 21 days pre-parturition to 14 days postpartum, spanning a total of 35 days. CONCLUSIONS: Serum anti-PEDV IgA antibody levels may serve as a valuable predictor for immunization effects, allowing for the assessment of colostrum IgA antibody levels up to 21 days in advance. This insight could enable veterinarians to timely adjust or optimize immunization strategies prior to parturition, thereby ensuring adequate passive immunity is conferred to piglets through colostral transfer postpartum.

Identification of factors affecting fattening efficiency of commercial pig herds and analysis of their impact at different performance levels.

Improving fattening efficiency is an important goal of breeding commercial pigs, especially for the large-scale pig farms. Fattening efficiency index (FEI) can be used to evaluate the fattening efficiency. The aim of this study was to identify the factors affecting the fattening efficiency of commercial pigs in large-scale pig farms and further study the impact of these factors on the production performance of commercial pig batches at different production levels. The data of 9,570 batches was mainly consisted of four parts (farm facilities, general information of piglets, production performance of nursery pigs and finishing pigs). A total of 28 variables were evaluated by the multi-variable linear regression models. The differences in production factors significantly correlated with FEI at piglets-finishing stage were compared among high-performing (HP), moderate-performing (MP), and low-performing (LP) batches of commercial pigs during the nursery and finishing stage. Among the 28 variables, 18 were significantly correlated with fattening efficiency (P < 0.05), including 11 continuous variables and seven discrete variables. The significant differences among the 11 consecutive variables in the HP, MP, and LP batches of commercial pigs mostly persisted from the piglets-nursery stage to the growing-finishing stage, ultimately affecting the FEI at piglets-finishing stage. For the seven significant discrete variables, the HP batches had a lower proportions in owned source of piglets, number of the purchasing piglets in spring and winter, number of batches in the East and North regions and five-way crossbred pigs, while a higher proportions in the use of closed circuit television video (CCTV) and wastes treatment system. The fattening efficiency of commercial pigs in large-scale pig farms was comprehensively affected by farm facilities, piglets, and production performance at nursery and finishing stage. The low fattening efficiency may have started at the end of nursery stage.

A duplex fluorescent quantitative PCR assay to distinguish the genotype I, II and I/II recombinant strains of African swine fever virus in China.

African swine fever (ASF) is a severe, hemorrhagic, and highly contagious disease caused by the African swine fever virus (ASFV) in both domestic pigs and wild boars. In China, ASFV has been present for over six years, with three genotypes of strains prevalent in field conditions: genotype I, genotype II, and genotype I/II recombinant strains. In order to differentiate among these three ASFV genotypes, a duplex fluorescent quantitative PCR method was established using specific probes and primers designed based on viral genes MGF_110-1L and O61R from ASFV strains reported in the GenBank database. Following optimization of reaction conditions, a duplex fluorescent quantitative PCR method was successfully developed. This method demonstrated no cross-reactivity with porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine reproductive and respiratory syndrome virus (PRRSV), classic swine fever virus (CSFV), porcine pseudorabies virus (PRV), porcine circovirus 2 (PCV2), porcine circovirus 3 (PCV3), highlighting its specificity. Sensitivity analysis revealed that the limits of detection (LODs) of this method were 2.95 × 10-1 copies/µL for the MGF_110-1L gene and 2.95 × 100 copies/µL for the O61R gene. The inter- and intra-group coefficients of variation were both <1%, indicating high reproducibility. In summary, the establishment of this duplex fluorescent quantitative PCR method not only addresses the identification of the ASFV recombinant strains but also allows for simultaneous identification of the three epidemic genotype strains.

Mobilizing endogenous neuroprotection: the mechanism of the protective effect of acupuncture on the brain after stroke.

Given its high morbidity, disability, and mortality rates, ischemic stroke (IS) is a severe disease posing a substantial public health threat. Although early thrombolytic therapy is effective in IS treatment, the limited time frame for its administration presents a formidable challenge. Upon occurrence, IS triggers an ischemic cascade response, inducing the brain to generate endogenous protective mechanisms against excitotoxicity and inflammation, among other pathological processes. Stroke patients often experience limited recovery stages. As a result, activating their innate self-protective capacity [endogenous brain protection (EBP)] is essential for neurological function recovery. Acupuncture has exhibited clinical efficacy in cerebral ischemic stroke (CIS) treatment by promoting the human body's self-preservation and "Zheng Qi" (a term in traditional Chinese medicine (TCM) describing positive capabilities such as self-immunity, self-recovery, and disease prevention). According to research, acupuncture can modulate astrocyte activity, decrease oxidative stress (OS), and protect neurons by inhibiting excitotoxicity, inflammation, and apoptosis via activating endogenous protective mechanisms within the brain. Furthermore, acupuncture was found to modulate microglia transformation, thereby reducing inflammation and autoimmune responses, as well as promoting blood flow restoration by regulating the vasculature or the blood-brain barrier (BBB). However, the precise mechanism underlying these processes remains unclear. Consequently, this review aims to shed light on the potential acupuncture-induced endogenous neuroprotective mechanisms by critically examining experimental evidence on the preventive and therapeutic effects exerted by acupuncture on CIS. This review offers a theoretical foundation for acupuncture-based stroke treatment.

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Acupuncture has a positive effect in the treatment of ischemic stroke (IS). A number of studies have confirmed that the role of acupuncture in the treatment of IS, which is closely related to its functions of regulating mitochondrial functions. In the present article, we review the mechanisms of acupuncture underlying improvement of mitochondria in the treatment of IS from 4 aspects： 1) protecting mitochondrial structure integrity, 2) regulative effect on mitochondrial functional activities, including regulating energy metabolism, reducing oxidative stress, suppressing calcium overload, and regulating mitochondrial membrane potential changes, 3) regulating mitochondrial quality control system, including promoting mitochondrial biosynthesis, regulating mitochondrial dynamics and apoptosis, and 4) regula-ting mitochondria-related apoptosis pathways. All of these may provide a theoretical basis for acupuncture in the treatment of IS and a reference for further research.

Comparative analysis of productive performance and fattening efficiency of commercial pigs in China for two consecutive years.

THE PURPOSE OF THIS STUDY: (1) propose an evaluation indicator of the fattening efficiency of commercial pigs (Yorkshire × Landrace × Duroc)-fattening efficiency index (FEI). (2) Analyze the correlation to find the main productive factors affecting the FEI. (3) Compare and analyze the yearly/monthly/different piglets' sources of productive performance in 2020 and 2021. The data included 2592 commercial pig batches in 2020 and 3266 in 2021, with a total of 6,134,234 commercial pigs. Descriptive statistics and difference analysis were carried out on 16 productive factors of a whole year and single/multiple sources for two consecutive years. The same period difference between the monthly data and the annual average were also analyzed. The top six productive factors correlated with FEI were average daily gain (ADG) (0.8080), feed conversion rate (FCR) (- 0.7203), survival rate (SR) (0.6968), number of deaths (- 0.4103), feeding days (- 0.3748) and body weight (BW) of marketing pigs (0.3369). The overall productive performance in 2021 was lower than that in 2020, which was reflected in more piglet sources and a lower BW of piglets, more deaths, a lower SR, longer feeding days, a lower ADG, a higher FCR and a lower FEI. The productive performance of a single source was better than that of multiple ones. The contrastive results of monthly data in 2020 and 2021 showed significant differences in most factors except for the number of marketing pigs, the number of piglets and feed consumption. The monthly trend of 15 factors for two consecutive years revealed similar trends only in the month of piglets purchasing, number of piglets sources, number of deaths and ADG. Compared with the annual average, the ADG significantly increased in May. The FEI of multiple sources was markedly lower than that of a single source. FEI may be suitable for evaluating the fattening efficiency of commercial pigs. The annual and monthly productive performance and fattening efficiency in 2021 were significantly lower than those in 2020. Single source was represented better productive performance and fattening efficiency than multiple ones.

Machine learning based personalized promotion strategy of piglets weaned per sow per year in large-scale pig farms.

BACKGROUND: The purpose of this study was to analyze the relationship between different productive factors and piglets weaned per sow per year (PSY) in 291 large-scale pig farms and analyze the impact of the changes in different factors on PSY. We chose nine different algorithm models based on machine learning to calculate the influence of each variable on every farm according to its current situation, leading to personalize the improvement of the impact in the specific circumstances of each farm, proposing a production guidance plan of PSY improvement for every farm. According to the comparison of mean absolute error (MAE), 95% confidence interval (CI) and R2, the optimal solution was conducted to calculate the influence of 17 production factors of each pig farm on PSY improvement, finding out the bottleneck corresponding to each pig farm. The level of PSY was further analyzed when the bottleneck factor of each pig farm changed by 0.5 standard deviation (SD). RESULTS: 17 production factors were non-linearly related to PSY. The top five production factors with the highest correlation with PSY were the number of weaned piglets per litter (WPL) (0.6694), mating rate within 7 days after weaning (MR7DW) (0.6606), number of piglets born alive per litter (PBAL) (0.6517), the total number of piglets per litter (TPL) (0.5706) and non-productive days (NPD) (- 0.5308). Among nine algorithm models, the gradient boosting regressor model had the highest R2, smallest MAE and 95% CI, applied for personalized analysis. When one of 17 production factors of 291 large-scale pig farms changed by 0.5 SD, 101 pig farms (34.7%) can increase 1.41 PSY (compared to its original value) on average by adding the production days, and 60 pig farms (20.6%) can increase 1.14 PSY on average by improving WPL, 45 pig farms (15.5%) can increase 1.63 PSY by lifting MR7DW. CONCLUSIONS: The main productive factors related to PSY included WPL, MR7DW, PBAL, TPL and NPD. The gradient boosting regressor model was the optimal method to individually analyze productive factors that are non-linearly related to PSY.

Study on the influence of different production factors on PSY and its correlation.

BACKGROUND: Finding out the key reproductive performance factors, affecting piglets weaned per sow per year (PSY) can improve the production efficiency and profitability of pig farms. The objective was to understand the actual distribution of different production factors and PSY of breeding pig farms, analyze the correlation to find the main production factors affecting PSY, and formulating a Production Efficiency Improvement Plan in practice. Data included 603 breeding pig farms from September 28, 2020 to September 26, 2021. Regression analysis was used to evaluate the relationship between PSY and key production factors, and the characteristics of total pig farms versus high performance (HP) pig farms (the production performance was in the top 10%) or top 5% pig farms were compared. Spearman's rank correlation coefficient was used to analyze the correlation between production factors and find the factors related to PSY. Non-linear support vector regression (NL-SVR) was used to analyze the personalized PSY improvement through a various change of the four key factors. RESULTS: The median distribution of 15 production factors and PSY in total pig farms were different from those of HP farms. All of data were distributed nonlinearly. Mating rate within 7 days after weaning (MR7DW), farrowing rate (FR), number of piglets born alive per litter (PBAL) and number of weaned piglets per litter (WPL) were moderately correlated with PSY, and the correlation coefficients were 0.5058, 0.4427, 0.3929 and 0.3839, respectively. When the four factors in NL-SVR changed in medium (0.5 piglet or 5%) or high level (1.0 piglet or 10%), PSY can be increased by more than 0.5. CONCLUSION: NL-SVR model can be used to analyze the impact of changes in key production factors on PSY. By taking measures to improve MR7DW, FR, PBAL and WPL, it may effectively improve the current PSY and fully develop the reproductive potential of sows.

Utilization and reproductive performance of gilts in large-scale pig farming system with different production levels in China: a descriptive study.

BACKGROUND: This study was to investigate the utilization and reproductive performance of gilts in large-scale pig farms. Data of this descriptive study included 169,013 gilts of 1540 gilts' batches on 105 large-scale pig farms from April 2020 to March 2021. According to the upper and lower 25th percentiles of piglets weaned per sow per year (PSY) during the research stage, pig farms were divided into three productivity groups: high-performing (HP), intermediate-performing (IP) and low-performing (LP) farms. On the basis of breeds, LP (LP-Total) farms was further divided into LP-breeding pig (LP-BP) and LP-commercial pig (LP-CP) groups. Average utilization, estrus and first mating data was collected from a total of 1540 gilts' batches. The age-related factors (introduction age, age at first estrus and age at first mating) and litter production (total number of piglets, number of piglets born alive and number of weaned piglets, as well as their proportion distribution) among HP and LP groups were compared. The litter production in different age groups were also analyzed. RESULTS: The introduction age, mortality and culling rate of HP farms were lower compared with LP farms. Total number of piglets per litter, number of piglets born alive per litter and number of weaned piglets per litter in HP farms were significantly more than those of LP groups, respectively. The proportion distribution peaks of litter production in HP farms were shifted about two more than those in LP groups, respectively; and the proportion of low litter production (eight per litter or less) was lower than that in LP groups. The results of different age groups showed that total number of piglets per litter and number of piglets born alive per litter in 220-279 d were the most, while that of 370 d was the least. CONCLUSIONS: The overall utilization and reproductive performance of gilts in HP farms was better than those of LP farms. The difference in utilization was reflected in introduction source, culling rate and mortality. While the age at first estrus and first mating, breeds and litter production were the main differences for reproductive performance.

Chemical Vapor Deposition of N-Doped Graphene through Pre-Implantation of Nitrogen Ions for Long-Term Protection of Copper.

Nitrogen-doped graphene (NG) was synthesized through the chemical vapor deposition (CVD) of graphene on Cu substrates, which were pre-implanted with N ions by the ion implantation method. The pre-implanted N ions in the Cu substrate could dope graphene by the substitution of C atoms during the CVD growth of graphene, forming NG. Based on this, NG's long-term protection properties for Cu were evaluated by ambient exposure for a corrosion test. The results showed that NG can obviously reduce the natural oxidation of Cu in the long-term exposure compared with the case of pristine graphene (PG) coated on Cu. Moreover, with the increase in pre-implanted N dose, the formed NG's long-term protection for Cu improved. This indicates that the modification of graphene by N doping is an effective way to improve the corrosion resistance of the PG coating owing to the reduction in its conductivity, which would inhibit galvanic corrosion by cutting off electron transport across the interface in their long-term protection. These findings provide insight into corrosion mechanisms of the graphene coating and correlate with its conductive nature based on heteroatoms doping, which is a potential route for improving the corrosion resistance of graphene as an effective barrier coating for metals.

Electroacupuncture Pretreatment Elicits Tolerance to Cerebral Ischemia/Reperfusion through Inhibition of the GluN2B/m-Calpain/p38 MAPK Proapoptotic Pathway.

Background: As one of the first steps in the pathology of cerebral ischemia, glutamate-induced excitotoxicity progresses too fast to be the target of postischemic intervention. However, ischemic preconditioning including electroacupuncture (EA) might elicit cerebral ischemic tolerance through ameliorating excitotoxicity. Objective: To investigate whether EA pretreatment based on TCM theory could elicit cerebral tolerance against ischemia/reperfusion (I/R) injury, and explore its potential excitotoxicity inhibition mechanism from regulating proapoptotic pathway of the NMDA subtype of glutamate receptor (GluN2B). Methods: The experimental procedure included 5 consecutive days of pretreatment stage and the subsequent modeling stage for one day. All rats were evenly randomized into three groups: sham MCAO/R, MCAO/R, and EA+MCAO/R. During pretreatment procedure, only rats in the EA+MCAO/R group received EA intervention on GV20, SP6, and PC6 once a day for 5 days. Model preparation for MCAO/R or sham MCAO/R started 2 hours after the last pretreatment. 24 hours after model preparation, the Garcia neurobehavioral scoring criteria was used for the evaluation of neurological deficits, TTC for the measurement of infarct volume, TUNEL staining for determination of neural cell apoptosis at hippocampal CA1 area, and WB and double immunofluorescence staining for expression and the cellular localization of GluN2B and m-calpain and p38 MAPK. Results: This EA pretreatment regime could improve neurofunction, decrease cerebral infarction volume, and reduce neuronal apoptosis 24 hours after cerebral I/R injury. And EA pretreatment might inhibit the excessive activation of GluN2B receptor, the GluN2B downstream proapoptotic mediator m-calpain, and the phosphorylation of its transcription factor p38 MAPK in the hippocampal neurons after cerebral I/R injury. Conclusion: The EA regime might induce tolerance against I/R injury partially through the regulation of the proapoptotic GluN2B/m-calpain/p38 MAPK pathway of glutamate.

Sunflower seed oil combined with ginseng stem-leaf saponins as an adjuvant to enhance the immune response elicited by Newcastle disease vaccine in chickens.

The present study was to evaluate the adjuvant effect of sunflower seed oil containing saponins extracted from the stem and leaf of Panax ginseng C.A. Meyer (E515-D) on the immune response induced by an inactivated Newcastle disease virus (NDV) in chickens. The results showed that E515-D promoted significantly higher serum NDV-specific HI and neutralizing antibody responses, IFN-γ and IL-4 levels, and lymphocyte proliferative responses to Con A, LPS, and NDV antigen than the conventional adjuvant Marcol 52. Different adjuvant effect between E515-D and Marcol 52 may be attributed to different genes expressed in two groups. Transcriptome analysis of splenocytes showed that there were 1198 differentially expressed genes (DEGs) with 539 up and 659 down regulated in E515-D group while 1395 DEGs with 697 up and 698 down regulated in Marcol 52 group in comparison with the control group. Analysis of gene ontology (GO) term and kyoto encyclopedia of Genes and Genomes (KEGG) pathways showed that the predominant immune related pathways included "Toll-like receptor signaling pathway", "NOD-like receptor signaling pathway", "C-type lectin receptor signaling pathway", and "Phosphatidylinositol signaling system" in E515-D group while Marcol 52 were "NOD-like receptor signaling pathway", "Phagosome", and "Lysosome", and the most relevant DEGs in E515-D group were STAT1, STAT2, PI3K, and IL-6. Considering the excellent adjuvant activity and vegetable origin, E515-D deserves further study as an adjuvant for vaccines used in food animals.

Enhanced Immune Responses with Serum Proteomic Analysis of Hu Sheep to Foot-and-Mouth Disease Vaccine Emulsified in a Vegetable Oil Adjuvant.

Our previous study demonstrated that a vegetable oil consisting of soybean oil, vitamin E, and ginseng saponins (SO-VE-GS) had an adjuvant effect on a foot-and-mouth disease (FMD) vaccine in a mouse model. The present study was to compare the adjuvant effects of SO-VE-GS and the conventional ISA 206 on an FMD vaccine in Hu sheep. Animals were intramuscularly (i.m.) immunized twice at a 3-week interval with 1 mL of an FMD vaccine adjuvanted with SO-VE-GS (n = 10) or ISA 206 (n = 9). Animals without immunization served as control (n = 10). Blood was sampled prior to vaccination and at 2, 4, 6, and 8 weeks post the booster immunization to detect FMD virus (FMDV)-specific IgG. Blood collected at 8 weeks after the booster was used for the analyses of IgG1 and IgG2, serum neutralizing (SN) antibody, IL-4 and IFN-γ production, and proteomic profiles. The results showed that IgG titers rose above the protection level (1:128) in SO-VE-GS and ISA 206 groups after 2 and 4 weeks post the booster immunization. At 6 weeks post the booster, the ISA 206 group had 1 animal with IgG titer less than 1:128 while all the animals in the SO-VE-GS group retained IgG titers of more than 1:128. At 8 weeks post the booster, 6 of 9 animals had IgG titers less than 1:128 with a protective rate of 33.3% in the ISA 206 group, while only 1 of 10 animals had IgG titer less than 1:128 with a protective rate of 90% in the SO-VE-GS group, with statistical significance. In addition, IgG1, IgG2, SN antibodies, IL-4, and IFN-γ in the SO-VE-GS group were significantly higher than those of the ISA 206 group. Different adjuvant effects of SO-VE-GS and ISA 206 may be explained by the different proteomic profiles in the two groups. There were 39 and 47 differentially expressed proteins (DEPs) identified in SO-VE-GS compared to the control or ISA 206 groups, respectively. In SO-VE-GS vs. control, 3 immune related gene ontology (GO) terms and 8 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were detected, while 2 immune related GO terms and 5 KEGG pathways were found in ISA 206 vs. control. GO and KEGG analyses indicated that 'positive regulation of cytokine secretion', 'Th1/Th2 cell differentiation', and 'Toll-like receptor signaling pathways', were obviously enriched in the SO-VE-GS group compared to the other groups. Coupled with protein-protein interaction (PPI) analysis, we found that B7TJ15 (MAPK14) was a key DEP for SO-VE-GS to activate the immune responses in Hu sheep. Therefore, SO-VE-GS might be a promising adjuvant for an FMD vaccine in Hu sheep.

A Solution with Ginseng Saponins and Selenium as Vaccine Diluent to Increase Th1/Th2 Immune Responses in Mice.

Pseudorabies is an important infectious disease of swine, and immunization using attenuated pseudorabies virus (aPrV) vaccine is a routine practice to control this disease in swine herds. This study was to evaluate a saline solution containing ginseng stem-leaf saponins (GSLS) and sodium selenite (Se) as a vaccine adjuvant for its enhancement of immune response to aPrV vaccine. The results showed that aPrV vaccine diluted with saline containing GSLS-Se (aP-GSe) induced significantly higher immune responses than that of the vaccine diluted with saline alone (aP-S). The aP-GSe promoted higher production of gB-specific IgG, IgG1, and IgG2a, neutralizing antibody titers, secretion of Th1-type (IFN-γ, IL-2, IL-12), and Th2-type (IL-4, IL-6, IL-10) cytokines, and upregulated the T-bet/GATA-3 mRNA expression when compared to aP-S. In addition, cytolytic activity of NK cells, lymphocyte proliferation, and CD4+/CD8+ ratio was also significantly increased by aP-GSe. More importantly, aP-GSe conferred a much higher resistance of mice to a field virulent pseudorabies virus (fPrV) challenge. As the present study was conducted in mice, further study is required to evaluate the aP-GSe to improve the vaccination against PrV in swine.

Immunomodulatory effect of thymopentin on lymphocytes from supramammary lymph nodes of dairy cows.

Previous study showed that injection of thymopentin (TP 5) in the area of supramammary lymph nodes (SMLN) had therapeutic effect on the intramammary infection (IMI) in cows. This study was to explore the underlying mechanisms by investigating the immunomodulatory effect of TP 5 on SMLN lymphocytes. Lymphocyte proliferation, cell cycle distribution and cytokine mRNA expression were determined by MTT, FCM and RT-qPCR, respectively. Laser scanning confocal microscope (LSCM) was used to observe the binding between TP 5 and SMLN lymphocytes. Moreover, RNA-sequencing (RNA-seq) was performed to observe the difference between the lymphocytes with and without TP 5 treatment. The results showed that TP 5 significantly promoted lymphocyte proliferation, accelerated cell cycle progression, and enhanced mRNA expression of IL-17A and IL-17F. Laser scanning confocal microscopic analysis revealed the binding of TP 5 to the surface of SMLN lymphocytes. A total of 1094 genes were identified as differentially expressed genes (DEGs) using RNA-seq with 692 up- and 402 down-regulated genes. 48 significantly enriched GO terms were identified by RNA-seq. In KEGG analysis, 1/3 of DEGs were enriched in the immune system pathway, including IL-17 signaling pathway, cytokine-cytokine receptor interaction, Th1 and Th2 cell differentiation, T cell receptor signaling pathway, Th17 cell differentiation. Among them, IL-17 signaling pathway was the most prominent. This study suggested that the therapeutic benefit of TP 5 in the treatment of bovine mastitis might be attributed to its immunomodulatory activity in SMLN lymphocytes.

Ginseng stem-leaf saponins in combination with selenium enhance immune responses to an attenuated pseudorabies virus vaccine.

Pseudorabies, a herpesvirus infection, is mainly controlled by using attenuated live vaccines. In this study, the effect of ginseng stem and leaf saponins (GSLS) in combination with selenium (Se; in the form of sodium selenite) on vaccination against attenuated pseudorabies virus (aPrV) was evaluated. It was found that GSLS and Se have an adjuvant effect and that a combination of GSLS and Se stimulates significantly enhanced immune responses than does GSLS or Se alone. Following oral administration of GSLS, mice immunized with an attenuated PrV vaccine diluted in Se-containing physiological saline solution (PSS) provoked a significantly stronger gB-specific serum antibodies response (IgG, IgG1 and IgG2a), enhanced lymphocyte proliferation and cytolytic activity of NK cells, along with higher production of cytokines (IFN-γ, IL-12, IL-5 and IL-10) by splenocytes. Notably, the combination of GSLS and Se conferred a much higher resistance to fPrV challenge after immunization of the mice with aPrV vaccine. This study offers convincing experimental evidence that an injection of Se with oral GSLS is a promising adjuvant combination that improves the efficacy of vaccination against PrV and deserves further study regarding improvement of responses to other animal vaccines.

Signaling pathway underlying splenocytes activation by polysaccharides from Atractylodis macrocephalae Koidz.

Previous study demonstrated that total polysaccharides isolated from Atractylodis macrocephalae Koidz. (RAMPtp) were effective to eliminate intramammary infection in cows. The present study was designed to investigate the immunomodulatory activity of RAMPtp in mouse splenocytes. Splenocyte proliferation, natural killer (NK) cytotoxicity, productions of NO and cytokines, transcription factor activity as well as the signal pathways and receptor were examined. The results showed that RAMPtp significantly promoted splenocyte proliferation and made the cells enter S and G2/M phases, increased ratios of T/B cells, boosted NK cytotoxicity, enhanced transcriptional activities of nuclear factor of activated T cells (NFAT), nuclear factor κB (NF-κB) and activator protein 1 (AP-1), and stimulated secretions of NO, immunoglobulin G (IgG) and multiple cytokine families (IL-1α, IL-1ß, IL-2, IL-3, IL-4, IL-6, IL-10, IL-12p40, IL-12p70, IL-13, IFN-γ, TNF-α, G-CSF, GM-CSF, KC, MIP-1α, MIP-1ß, RANTES and Eotaxin). In addition, all the specific inhibitors against the mitogen-activated protein kinases (MAPKs) and NF-κB significantly suppressed the IL-6 production induced by RAMPtp. Moreover, splenocytes from Toll-like receptor 4 (TLR4) deficient mouse responded equally to RAMPtp stimulation as the wild-type. Therefore, RAMPtp might induce splenocytes activation at least in part via the TLR4-independent MAPKs and NF-κB signaling pathways. The present results would be useful to further understand the immunomodulatory mechanisms of RAMPtp in elimination of intramammary infection in cows.

Soybean oil containing ginseng saponins as adjuvants promotes production of cytokines and enhances immune responses to foot-and-mouth disease vaccine.

In the present study, the adjuvant effect of soybean oil containing ginseng root saponins (SO-GS-R) on the immune response to foot-and-mouth disease vaccine (FMDV) in mice was investigated. When immunized with FMDV antigen emulsified in an SO-GS-R formulation, mice generated remarkably higher serum antibody and cytokine responses than mice immunized with FMDV antigen alone. To elucidate the mechanisms underlying the adjuvant effect of SO-GS-R, we measured cytokines in serum and muscle tissue after intramuscular injection of SO-GS-R. The results showed that injection of SO-GS-R significantly increased the levels of IL-1ß, IL-5, IL-6, G-CSF, KC, MCP-1, MIP-1α, and MIP-1ß in both serum and muscle. These results suggested that SO-GS-R recruits neutrophils, eosinophils, T cells and macrophages, causing immune cell recruitment at the injection site, driving antigen-presenting cells to actively participate in the onset of immunity, and amplifying the immune responses. Considering its adjuvant activity and plant-derived properties, SO-GS-R should be further studied for its adjuvant effect on vaccines used in food animals.

Structural analysis and immunomodulatory effect of polysaccharide from Atractylodis macrocephalae Koidz. on bovine lymphocytes.

RAMP60, RAMP70, RAMP80 and RAMPtp extracted from the rhizome of Atractylodis macrocephalae Koidz. were chemically analyzed and tested for their lymphocyte stimulation. Among them, only RAMPtp was found to induce proliferation of lymphocytes from bovine supramammary lymph node (SMLN). It was found that RAMPtp was homogeneous polysaccharide with Mn of 1.867×103 by GPC analysis, composed of glucose, mannose, rhamnose, arabinose and galactose, with mass percentages of 60.67%, 14.99%, 10.61%, 8.83% and 4.90%, respectively by HPLC analysis, connected by 1, 3-linked ß-D Galp and 1, 6-linked ß-D Galp residues by NMR analysis and was amorphous in structure by SEM analysis. When SMLN lymphocytes were cultured with RAMPtp in vitro, increased [Ca2+]i, more cells in S and G2/M phases and upregulated IFN-γ and IL-17A while downregulated IL-4 mRNA expressions as well as the binding between SMLN lymphocytes and RAMPtp were observed. These findings might explain the lymphocyte-stimulating property of RAMPtp.

Efficacy of vaccination and nisin Z treatments to eliminate intramammary Staphylococcus aureus infection in lactating cows.

This study evaluated the effect of a Staphylococcus aureus bacterin and nisin on bovine subclinical mastitis. A total of 75 Holstein subclinically mastitic cows were randomly allocated to three groups with 25 cows per group. In group I, an intramammary infusion of nisin Z at a dose of 2.50×106 IU was carried out once daily for three days, and an autogenous S. aureus bacterin was inoculated into the supramammary lymph node one week before and one week after nisin treatment. In group II, nisin was administered in the same way as in group I, but no bacterin was inoculated. Group III received no treatment and served as a control. Milk was aseptically sampled from the affected quarters before and 2, 4, and 6 weeks after treatment, for bacteriological examination and analyses of N-acetyl-ß-D-glucosaminidase (NAGase) activity, somatic cell count (SCC), and milk protein and fat contents. Results indicated that, compared to the nisin-treated group, nisin-bacterin treatment significantly reduced intramammary S. aureus infections, reduced the number of quarters with milk SCCs of more than 5×105 cells/ml, and increased the protein and fat contents of the milk. Therefore, nisin-bacterin therapy is suggested when subclinical mastitis occurs in lactating cows.