The first molecular detection of benzimidazole resistance in Haemonchus contortus from sheep in Hejing and Minfeng counties of Southern Xinjiang.

To understand the benzimidazole (BZ) resistance of Haemonchus contortus in Southern Xinjiang, three single nucleotide polymorphisms (SNPs) designated as F167Y, E198A, and F200Y, in the isotype-1 ß-tubulin gene which are associated with BZ resistance, were investigated for H. contortus populations from sheep in Hejing and Minfeng counties of Southern Xinjiang. In brief, a total of 190 H. contortus adults were collected from 52 out of 70 slaughtered sheep in city abattoirs across two regions in Southern Xinjiang. The species identity of each adult worm was confirmed by PCR amplification of ITS-2 using H. contortus-specific primers targeting the ITS-2. The samples were then investigated for BZ-related SNPs at locus 167, 198, and 200, by PCR-sequencing of the isotype-1 ß-tubulin gene. The results showed that only E198A and F200Y mutations were detected in the investigated H. contortus populations. The E198A mutation (homozygous and heterozygote resistant: found in 40% and 30% of sequenced samples from Minfeng and Hejing counties, respectively) was predominant compared with the F200Y mutation (homozygous and heterozygote resistant: found in 14% and 13.3% of sequenced samples from Minfeng and Hejing counties, respectively). The results indicate a high prevalence of BZ resistance in H. contortus populations from certain areas of Southern Xinjiang. Our findings provide valuable information for the prevention and control of H. contortus in areas with similar conditions.

Extraction and immunomodulatory effects of acid Lagenaria siceraria (Molina) Standl. Polysaccharide on chickens.

Herbal polysaccharides are extensively studied as vaccine adjuvants due to their safety and potent immunoenhancing activity. This study aimed to analyze the structure of Lagenaria siceraria (Molina) Standl polysaccharide (LSP50) and investigate its adjuvant activity for the H9N2 vaccine in broiler chickens. Structural analysis revealed that LSP50 primarily consisted of rhamnose, arabinose, xylose, mannose, glucose, and galactose with molar ratios of 23.12: 12.28: 10.87: 8.26: 2.64: 22.82 respectively. The adjuvant activity of LSP50 was evaluated, which showing significant enhancements compared to the H9N2 group. Parameters including the immune organ index, H9N2 specific IgG level, cytokines contents (IFN-γ, IL-2, IL-4, and IL-5), and the proportion of CD3e+CD8aT+cells were significantly increased in the LSP50 group (P < 0.05). Additionally, sequencing results showed that LSP50 modulates the immune response by regulating PLA2G12B and PTGDS genes involved in the arachidonic acid pathway. These findings were further validated through qPCR analysis to affirm the reliability of the sequencing data. In conclusion, our results demonstrate that LSP50 exhibits potent adjuvant activity, enhancing both cellular and humoral immunity.

Effects of Alhagi maurorum Medik polysaccharide derived from different regions on the intestinal immune functions of lambs.

Introduction: Plant polysaccharide are widely studied as potential prebiotics because of their potential to protect and enhance the immunity of lambs. Methods: In this study, the polysaccharide content of Alhagi maurorum Medik from Aksu (AK) and Shanshan (SS) at different cutting periods was determined, and the functions of Alhagi maurorum Medik polysaccharide were investigated to useas an immunomodulator. Results: Our results indicated that the content of Alhagi maurorum Medik polysaccharide is the highest at the maturity stage, and the polysaccharide content of Alhagi maurorum Medik produced in Shanshan area is higher as compared to the Aksu area. The serum IgG, duodenum IgA, TNF-α, IL-4, IL-10 contents, jejunum IgA, TNF-α, IL-4, IL-17 contents, ileum IgA, IL-17 contents, duodenum villus height, crypt depth and jejunum crypt depth of lambs were significantly adjusted in the SS group as compared to CK control group and AK groups (p < 0.05). Furthemore, the sequencing results showed that SS polysaccharide promoted the release of large amounts of IgA and enhanced the immunal function of intestine by regulating the IgA production pathway and B-cell receptor signaling to activate B cells in the T-dependent pathway. Discussion: Altogether, Alhagi maurorum Medik polysaccharide from SS group holds a promising potential to be used as a valuable immunopotentiator for optimizing the immune system of intestine in lambs.

Effects of Alhagi camelorum Fisch polysaccharide from different regions on growth performance and gastrointestinal microbiota of sheep lambs.

Polysaccharides derived from Alhagi camelorum Fisch possess diverse activities, making them a potential prebiotic candidates for enhancing lamb health. This study investigated the immunomodulatory effects of Alhagi camelorum Fisch polysaccharides from Aksu (AK) and Shanshan (SS) regions on sheep lambs. The results showed that sheep lambs in the SS group exhibited significantly increased (p < 0.05) average daily gain, levels of growth hormone (GH), insulin (INS), IgA and IgM, and cytokines IL-4, IL-10, IL-17, TNF-α and IFN-γ compared to those in the control check (CK) group. Moreover, the SS treatment significantly increased the diversity and abundance of beneficial bacteria, while concurrently diminishing the prevalence of harmful bacteria. Additionally, it modulated various metabolic pathways, promoted lamb growth, improved immunity, reduced the risk of gastrointestinal disease and improved the composition of gastrointestinal microbiota. In summary, our findings highlight the potential of SS treatment in enhancing gastrointestinal health of sheep lambs by improving intestinal function, immunity, and gut microbiome. Consequently, these results suggest that Alhagi camelorum Fisch polysaccharides derived from Shanshan regions holds promising potential as a valuable intervention for optimizing growth performance in sheep lambs.

Structure characterization and intestinal immune promotion effect of polysaccharide purified from Alhagi camelorum Fisch.

This study investigated the structure of acid Alhagi camelorum Fischa polysaccharide (aAP) and its impact on intestinal activity in mice. The results showed that aAP comprised of the fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, galacturonic acid, glucuronic acid with the molar ratio of 0.81:14.97:10.84:11.14:3.26:0.80:0.80:54.92:2.47 with the molecular weight (Mw) of 22.734 kDa. Additionally, the composition of aAP was assessed via FT-IR, methylation, and NMR analyses, indicating that the backbone of the aAP was consisted of →4)-α-D-GalpA-6-OMe-(1 â†’ 4)-α-GalpA-(1 â†’ and →4)-α-D-GalpA-6-OMe-(1 â†’ 2)-α-L-Rhap-(1→, as well as →4)-ß-D-Galp- and →5)-α-L-Araf- for the branched chain. Furthermore, ICR mice underwent intragastric administration of different concentrations of aAP for 7 consecutive days. The results showed that aAP enhanced the murine spleen and thymus indices, promoted the secretion of serum lgG antibody, intestinal lgA antibody and intestinal cytokines, improved the morphology of intestinal villi and crypts, enhanced quantity of intestinal IELs and IgA+ cells, and activated T lymphocytes and DC cells in MLNs. In summary, these findings suggest that the utilization of aAP could enhance the immune response of the murine intestinal mucosa.

Preparation and activity study of Ruoqiang jujube polysaccharide copper chelate.

Background: Polysaccharide metal chelate exhibit both immunoregulatory activity and metal element supplementation effects. Methods: In this study, Ruoqiang jujube polysaccharide copper chelate (RJP-Cu) was prepared and the preparation conditions were optimized using the response surface method. Subsequently, RJP-Cu was administered to lambs to evaluate its impact on growth performance, copper ion (Cu2+) supplementation, immune enhancement, and intestinal flora was evaluated. Results: The results indicated that optimal RJP-Cu chelation conditions included a sodium citrate content of 0.5 g, a reaction temperature of 50°C, and a solution pH of 8.0, resulting in a Cu2+ concentration of 583°mg/kg in RJP-Cu. Scanning electron microscopy (SEM) revealed significant structural changes in RJP before and after chelation. RJP-Cu displaying characteristic peaks of both polysaccharides and Cu2+ chelates. Blood routine indexes showed no significant differences among the RJP-Cu-High dose group (RJP-Cu-H), RJP-Cu-Medium dose group (RJP-Cu-M), RJP-Cu-low dose group (RJP-Cu-L) and the control group (p > 0.05). However, compared with the control group, the RJP-Cu-H, M, and L dose groups significantly enhanced lamb production performance (p < 0.05). Furthermore, RJP-Cu-H, M, and L dose groups significantly increased serum Cu2+ concentration, total antioxidant capacity (T-AOC), catalase (CAT), and total superoxide dismutase (T-SOD) contents compared with control group (p < 0.05). The RJP-Cu-H group exhibited significant increases in serum IgA and IgG antibodies, as well as the secretion of cytokines IL-2, IL-4, and TNF-α compared to the control group (p < 0.05). Furthermore, RJP-Cu-H group increased the species abundance of lamb intestinal microbiota, abundance and quantity of beneficial bacteria, and decrease the abundance and quantity of harmful bacteria. The RJP-Cu-H led to the promotion of the synthesis of various Short Chain Fatty Acids (SCFAs), improvements in atrazine degradation and clavulanic acid biosynthesis in lambs, while reducing cell apoptosis and lipopolysaccharide biosynthesis. Conclusion: Thus, these findings demonstrate that RJP-Cu, as a metal chelate, could effectively promote lamb growth performance, increase Cu2+ content, and potentially induce positive immunomodulatory effects by regulating antioxidant enzymes, antibodies, cytokines, intestinal flora, and related metabolic pathways.

Preparation and sulfate modified of Lagenaria siceraria (Molina) Standl polysaccharide and its immune-enhancing adjuvant activity.

Herbal polysaccharides and their modifiers used as vaccine adjuvants have been widely investigated due to their safety and good immunoenhancing activity. In this study, the 50% ethanol concentration precipitated Lagenaria siceraria(Molina) standl polysaccharide (LSP50) and sulfated modified LSP50 (sLSP50) was prepared, and their characterization was investigated. LSP50 and sLSP50-1.5 were used as vaccine adjuvants to immunize chickens, and the strength and type of immune responses induced by different adjuvants were detected. Our results showed that LSP50 was homogeneous polysaccharides, and the carbohydrate content was 98.6%. The sLSP50-1.5 with the DS value of 1.5 was optimized by response surface methodology. The sLSP50-1.5 has both characteristics of polysaccharide functional groups and sulfate functional groups. Adjuvant activity of LSP50 and sLSP50-1.5 showed that LSP50 and sLSP50-1.5 could induce long-lasting and high hemagglutination (HI) titers, antigen-specific lgG-NDV antibody, splenic lymphocyte proliferation, high immune organ index. Moreover, chicken immunized with sLSP50-1.5 showed a strong mixed Th1-type (IFN-γ and TNF-α) and Th2-type (IL-4 and IL-6) cytokines expression. Thus, these findings demonstrated that sLSP50-1.5 as a vaccine adjuvant can induce a mixed cellular and humoral immune response and can potentially serve as an effective vaccine adjuvant for NDV antigen.

Polyethyleneimine modified Pickering emulsion as a novel adjuvant to induce strong and long-lasting immune responses.

Poly (lactic-co-glycolic acid) (PLGA) nanoparticles are widely-investigated vaccine adjuvants owing to their safety, antigen slow-release ability, and good adjuvants activity. This study involved the preparation of the polyethyleneimine-modified immunopotentiator Alhagi honey polysaccharide encapsulated PLGA nanoparticles (PEI-AHPP) and the assembly of the Pickering emulsion with PEI-AHPP as shell and squalene as core (PEI-PPAS). Furthermore, PEI-AHPP and PEI-PPAS were characterized. To assess the strength and type of immune responses induced by different adjuvants, the chickens were immunized with H9N2-absorbed nanoparticle formulations. Our results showed that since the PEI-PPAS possess rough strawberry-like surfaces, a large number of antigens can be absorbed on their surfaces through simple mixing. Compared to PEI-AHPP, PEI-PPAS was found to exhibit better stability and antigen-loading efficiency. The adjuvant activity of the nanoparticles showed PEI-PPAS/H9N2 to induce long-lasting and high Hemagglutination inhibition (HI) titers, high thymus, spleen, and organ index of the bursa of Fabricius. Moreover, the chickens immunized with PEI-PPAS/H9N2 showed a mixture of high CD4+ and CD8a+ T-cells in the spleen and strong Th1 and Th2-type cytokines secretion. Thus, these findings demonstrated PEI-PPAS to be a vaccine adjuvant inducing a mixed cellular and humoral immune response, which can potentially serve as an effective vaccine delivery adjuvant system for the H9N2 antigen.

Poly (lactic-co-glycolic acid) nanoparticle-based vaccines delivery systems as a novel adjuvant for H9N2 antigen enhance immune responses.

Poly (lactic-co-glycolic acid) (PLGA) nanoparticle used as vaccine adjuvants have been widely investigated due to their safety, antigen slow-release ability, and good adjuvants activity. In this study, immunopotentiator Alhagi honey polysaccharide encapsulated PLGA nanoparticles (AHPP) and assembled pickering emulsion with AHPP as shell and squalene as core (PPAS) were prepared. Characterization of AHPP and PPAS were investigated. H9N2 absorbed nanoparticles formulations were immunized to chicken, then the magnitude and kinetics of antibody and cellular immune responses were assessed. Our results showed that PPAS had rough strawberry-like surfaces, a large number of antigens could be absorbed on their surfaces through simple mixing. Adjuvant activity of PPAS showed that, PPAS/H9N2 can induce long-lasting and high HI titers, high thymus, spleen, and bursa of fabricius organ index. Moreover, chicken immunized with PPAS/H9N2 showed a mixed high differentiation of CD4+ and CD8a+ T cell, and strong Th1 and Th2-type cytokines mRNA expression. Thus, these findings demonstrated that PPAS could induce a strong and long-term cellular and humoral immune response, and has the potential to serve as an effective vaccine delivery adjuvant system for H9N2 antigen.

Traditional Uyghur Medicine: Concepts, Historical Perspective, and Modernization.

Context • Traditional Uyghur medicine (TUM) is rooted in ancient Uyghur medical theory that was developed with the combined essence of different traditional medicines, such as Han Chinese, Egyptian, ancient Greek, Arabian, Persian, and Indian medicines. Modern experimental methods and technologies for disease diagnoses have accelerated the modernization of Uyghur medicine. Objective • The research team intended to compile a comprehensive introduction to TUM and to determine the current state of research in the field to establish a basis for future modernization of Uygur medicine. Design • The research team collected information from several databases-the China National Knowledge Infrastructure, the Wanfang Databases, and PubMed-as well as from Uyghur medical books. They also interviewed Uyghur medical scholars to ensure the authenticity of the Uyghur medical theory presented. The registry database of the China Food and Drug Administration was also used to search for and screen registered TUMs. Setting • The selection of articles and further inclusion in the review was performed in the College of Veterinary Medicine, Northwest A&F University (Yangling, China). Results • TUM has been developed to a unique, comprehensive theoretical system with the concepts and theories for clinical diagnosis, treatment, and medication. Advancements of modern experimental methods and disease diagnoses have accelerated the modernization of Uyghur medicine. The establishments of a series of standards/regulations legalize Uyghur drug production, supervision, and management. Conclusions • The future development of Uygur medicine should begin with the standardization of planting, production, and laboratory and clinical practices to form a complete system with the support and participation of the government to realize the modernization of TUM finally worldwide. One pressing matter is a full analysis of the requirements and standards of the dominant international pharmaceutical markets as applied to natural medicinal preparations, including TUM preparations. Knowledge of the exact curative effects of Uygur medicine and the development of TUM preparations that conform to international standards would enable them to be better accepted by the mainstream market.

The immunological adjuvant activity of gypenosides liposome against Newcastle disease vaccine.

The adjuvant activity of gypenosides liposome (GPSL) encapsulated with liposome was investigated in vitro and in vivo. In vitro, different concentrations of GPSL were added into chicken's peripheral blood lymphocytes and splenic lymphocyte. The results showed that GPSL could significantly enhance T and B lymphocytes proliferation singly or synergistically with PHA and LPS and the efficacy were superior to those of gypenosides (GPS) and blank liposome (BL) at most of concentrations. In vivo, three hundred and fifty 14-day-old chickens were assigned to 7 groups randomly and vaccinated with Newcastle disease (ND) vaccine. Simultaneously, the chickens in experimental groups were, respectively, oral administration with the GPSL at three doses, GPS and BL. The results showed that GPSL could significantly enhance lymphocyte proliferation, increase antibody titer, and promote cytokine secretion in vitro and in vivo, moreover, the adjuvant activity of GPSL was better than those of GPS and BL. These indicated that formulations of GPS and liposome can further enhance the immune response against ND vaccine compared with the adjuvant alone.

Optimization on preparation condition of propolis flavonoids liposome by response surface methodology and research of its immunoenhancement activity.

The aim of this study is to prepare propolis flavonoids liposome (PFL) and optimize the preparation condition and to investigate further whether liposome could promote the immunoenhancement activity of propolis flavonoids (PF). PFL was prepared with ethanol injection method, and the preparation conditions of PFL were optimized with response surface methodology (RSM). Moreover, the immunoenhancement activity of PFL and PF in vitro was determined. The result showed that the optimal preparation conditions for PFL by response surface methodology were as follows: ratio of lipid to drug (w/w) 9.6 : 1, ratio of soybean phospholipid to cholesterol (w/w) 8.5 : 1, and speed of injection 0.8 mL·min(-1). Under these conditions, the experimental encapsulation efficiency of PFL was 91.67 ± 0.21%, which was close to the predicted value. Therefore, the optimized preparation condition is very reliable. Moreover, the results indicated that PFL could not only significantly promote lymphocytes proliferation singly or synergistically with PHA, but also increase expression level of IL-2 and IFN-γ mRNA. These indicated that liposome could significantly improve the immunoenhancement activity of PF. PFL demonstrates the significant immunoenhancement activity, which provides the theoretical basis for the further experiment in vivo.

In vitro antiviral activity of sulfated Auricularia auricula polysaccharides.

Total Auricularia auricula polysaccharide (AAP(t)) was prepared by extracting and removing the proteins. Column chromatography was used to further graded it into AAP(1) and AAP(2). Three AAPs were modified by chlorosulfonic acid-pyridine method to obtain three sulfated AAPs (sAAPs), sAAP(t), sAAP(1) and sAAP(2), respectively. Three sAAPs and Newcastle disease virus (NDV) were added into cultivation system of chicken embryo fibroblast (CEF) in three manners, pre-, post- and simultaneous-adding polysaccharide with NDV respectively, taking three non-modified AAPs as control. Their anti-viral activities were compared by MTT method. The results showed that sAAPs and AAPs at a certain concentration could significantly inhibit the cellular infectivity of NDV in three manners. The effects of sAAPs were better than that of AAPs. It indicated that sulfated modification could enhance the antiviral activity of AAP. sAAP(1) and sAAP(t) possessed stronger activity and would be as the component of a new-type antiviral drug.

Immuno-enhancing activity of sulfated Auricularia auricula polysaccharides.

The crude total Auricularia auricula polysaccharide (AAPct) was extracted by water decoction and ethanol precipitation, protein was removed to obtain total A. auricula polysaccharide (AAPt), then was graded into AAP1 and AAP2 through column chromatography. sAAPt, sAAP1 and sAAP2 were prepared by chlorosulfonic acid-pyridine method. In vitro test, the effects of sAAPt, sAAP1, sAAP2, AAPt, AAP1 and AAP2, on chicken peripheral lymphocytes proliferation were compared. The results showed that sAAPt and sAAP1 demonstrated better effect. In vivo test, 14-day-old chickens were injected respectively with sAAPt, sAAP1, AAPt and AAP1 at the first vaccination of ND vaccine, once a day for three days. On days 7, 14, 21 and 28 after the first vaccination, the peripheral lymphocytes proliferation and antibody titer were determined. The results indicated that sAAPt possessed the best efficacy and would be expected to be used as a component of a new-type immunopotentiator.

Optimization of sulfated modification conditions of tremella polysaccharide and effects of modifiers on cellular infectivity of NDV.

Based on our previous research, sulfated modification conditions of Tremella polysaccharide (TPS), the chlorosulfonic acid to pyridine (CSA-Pry) ratio, reaction temperature and time, were optimized by L(9) (3(4)) orthogonal design taking the yield and degree of sulfation (DS) of modifiers as indexes. Two TPSs, TPS(tp) and TPS(70c), were modified under optimized conditions. The effects of two modifiers, sTPS(tp) and sTPS(70c), on cellular infectivity of NDV were determined by MTT method taking the non-modified TPS(tp), TPS(tc) and TPS(70c) as controls. The results showed that the optimized modification conditions were reaction temperature of 80°C, CSA-Pry ratio of 1:6 and reaction time of 1.5h. Five polysaccharides at proper concentrations could significantly inhibit the infectivity of NDV to CEF. The virus inhibitory rates of sTPS(tp) at 1.563 µg mL(-1) group were the highest and significantly higher than those of other three non-modified polysaccharide groups in three sample-adding modes. This indicated that sulfated modification could significantly improve the antiviral activity of TPS. sTPS(tp) possessed the best efficacy and would be as a component of antiviral polysaccharide drug.

Sulfated modification can enhance the immune-enhancing activity of lycium barbarum polysaccharides.

In test in vitro, four sulfated lycium barbarum polysaccharides (sLBPSs) with different degrees of sulfation (DS), sLBPS0.7, sLBPS1.1, sLBPS1.5 and sLBPS1.9, were added into cultured chicken peripheral lymphocytes and the changes of lymphocytes proliferation were compared by MTT assay taking the non-modified LBPS as control. Two sLBPSs with better efficacy, sLBPS1.5 and sLBPS1.9 were selected. In test in vivo, one hundred 14-day-old chickens were averagely divided into five groups randomly. The chickens except blank control group were vaccinated with Newcastle disease vaccine, repeated vaccination at 28 days old. At the same time of the first vaccination, the chickens in three experimental groups were injected with 0.5 mL of sLBPS1.5, sLBPS1.9 and LBPS at 4 mg mL(-1), in vaccination control group, with 0.5 mL of physiological saline, once a day for three successive days. On days 7, 14, 21 and 28 after the first vaccination, the changes of peripheral lymphocytes proliferation and serum HI antibody titer were determined. The result showed that two sLBPSs could significantly promote lymphocytes proliferation and enhance serum antibody titer. These results indicated that sulfated modification could enhance the immune-enhancing activity of LBPS, which there was a certain relativity with the DS of sulfated polysaccharide. sLBPS1.5 possessed the best efficacy and would be expected as the component drug of a new-type immunopotentiator.

Lycium barbarum polysaccharide inhibits the infectivity of Newcastle disease virus to chicken embryo fibroblast.

Lycium barbarum polysaccharide (LBPS) was extracted by water decoction and ethanol precipitation. After purification, four sulfated lycium barbarum polysaccharides (sLBPSs), sLBPS(0.7), sLBPS(1.1), sLBPS(1.5) and sLBPS(1.9), were prepared by chlorosulfonic acid-pyridine method respectively at four designed modification conditions. Four sLBPSs at 5 concentrations, within the safety concentration scope, and Newcastle disease virus (NDV) were added into cultivating system of chick embryo fibroblast (CEF) respectively in three modes, pre- and post-adding polysaccharide and simultaneous adding polysaccharide and virus after being mixed. The effects of sLBPSs on cellular infectivity of NDV were assayed by MTT method taking the non-modified LBPS as control. The results showed that sLBPS(1.5), sLBPS(1.9) and sLBPS(1.1) in three sample-adding modes, sLBPS(0.7) in simultaneous adding after being mixed could significantly inhibit the infectivity of NDV to CEF. The viral inhibitory rate of sLBPS(1.5) in pre- and simultaneous adding and sLBPS(1.9) in post-adding was the highest. Non-modified LBPS did not present significant effect in any sample-adding mode. These results indicated that sulfated modification could significantly enhance the antiviral activity of LBPS, which was correlated with the degree of sulfation (DS) of sLBPS. sLBPS(1.5) and sLBPS(1.9) possessed better activity and would be as the compositions of antiviral prescription.