Feeding Eucommia ulmoides extract enhances protection against high-temperature stress in chicks.

Chick's susceptibility to heat stress often leads to growth retardation, immune function impairment, disease, and mortality. This thesis explores the potential ameliorative effect of 0.8% Eucommia ulmoides extract (EUE) into the diet of heat-stressed chicks in a 15-d feeding trial. The investigation reveals that feeding EUE significantly enhances the BW, ADG, AFI, and F/G of chicks experiencing heat stress. Additionally, the EUE groups exhibited higher levels of T-AOC (at 7 and 15d), SOD (at 15 d), GSH-Px (at 15 d), as well as lower MDA concentrations (at 7 and 15d) in chick serum. Pathological changes and H&E staining revealed that EUE effectively improved tissue damage in the duodenum, heart, and stomach induced by heat stress in the chicks. The EUE groups also showed higher levels of IgA (at 7 d), IgG and IgM (at 7 and 15 d). RNA-seq and WGCNA analysis revealed that EUE mitigates cellular damage and losses in heat-stressed chicks primarily through pathways involving signal transduction, protein synthesis and degradation, as well as cell cycle regulation, particularly the latter. This investigation serves as a fundamental and cognitive framework for the development and application of Eucommia ulmoides feed additives aimed at safeguarding the well-being of chicks in adverse environmental conditions.

Overexpression of the NbZFP1 encoding a C3HC4-type zinc finger protein enhances antiviral activity of Nicotiana benthamiana.

Viral diseases are crucial determinants affecting tobacco cultivation, leading to a substantial annual decrease in production. Previous studies have demonstrated the regulatory function of the C3HC4 family of plant zinc finger proteins in combating bacterial diseases. However, it remains to be clarified whether this protein family also plays a role in regulating resistance against plant viruses. In this study, the successful cloning of the zinc finger protein coding gene NbZFP1 from Nicotiana benthamiana has been achieved. The full-length coding sequence of NbZFP1 is 576 bp. Further examination and analysis of this gene revealed its functional properties. The induction of NbZFP1 transcription in N. benthamiana has been observed in response to TMV, CMV, and PVY. Transgenic N. benthamiana plants over-expressing NbZFP1 demonstrated a notable augmentation in the production of chlorophyll a (P < 0.05). Moreover, NbZFP1-overexpressing tobacco exhibited significant resistance to TMV, CMV, and PVY, as evidenced by a decrease in virus copies (P < 0.05). In addition, the defense enzymes activities of PAL, POD, and CAT experienced a significant increase (P < 0.05). The up-regulated expression of genes of NbPAL, NbNPR1 and NbPR-1a, which play a crucial role in SA mediated defense, indicated that the NbZFP1 holds promise in enhancing the virus resistance of tobacco plant. Importantly, the results demonstrate that NbZFP1 can be considered as a viable candidate gene for the cultivation of crops with enhanced virus resistance.

Isolation and In Vitro cultivation of Lawsonia intracellularis from China.

Lawsonia intracellularis is an obligate intracellular bacterium that cannot be cultured by conventional bacteriological methods. Pigs infected with L. intracellularis suffer from decreased daily weight gain and poor feed conversion ratio. China is a large producer of pigs, but epidemiological investigation data of L. intracellularis has not been obtained in recent years. Additionally, there is no information about a L. intracellularis strain being successfully isolated and established in cell culture in China, and the above shortcomings limit understanding of the pathogenesis of L. intracellularis and alternative prevention and control methods. The aims of this study were to estimate the seroprevalence of L. intracellularis antibodies in eight major pig-producing provinces in China during 2019-2020, to isolate L. intracellularis from infected intestines and then to establish an infection model of L. intracellularis in mice. Our results showed that of the 3586 serum samples, 2837 (79.1%, 95% CI: 77.7%, 80.4%) were seropositive for the L. intracellularis antibody. Subsequently, the L. intracellularis strain LJS19051 from China was successfully isolated and established in cell culture. Furthermore, L. intracellularis DNA and antibodies could be detected in the feces and serum samples of infected mice, respectively. Moreover, infected crypts showed typical proliferative enteropathies (PE) lesions and L. intracellularis antigen was detected in infected mice by immunofluorescence at 28 days post inoculation. The results indicated that the new L. intracellularis strain LJS19051 was obtained and could successfully proliferate in ICR mice.

Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay.

Proliferative enteropathy (PE) is an infectious enteric disease caused by Lawsonia intracellularis (L. intracellularis) and is endemic in pig herds worldwide. However, a L. intracellularis-specific monoclonal antibody plays an important role in the evaluation of L. intracellularis infection in vitro. Therefore, the objective of this study was to produce and identify the characteristics of a new monoclonal antibody against the outer membrane protein (Omp2) of L. intracellularis and apply it in an indirect immunofluorescence assay (IFA) and immunocytochemistry (IHC). The results indicated that three highly specific monoclonal antibodies against the Omp2 protein (4D9, 3G2, and 7G5) of L. intracellularis were obtained by using purified Omp2 as an immunogen, the titers of ascitic fluids of 4D9, 3G2, and 7G5 cells were 1:2,048,000, 1:512,000, and 1:256,000, respectively. IFA analysis showed that the 4D9, 3G2, and 7G5 have no cross-reactivity with other enteric bacteria commonly found in the ilea of pigs or closely related to L. intracellularis, such as Desulfovibrio, Bilophila wadsworthia (B. wadsworthia), Salmonella choleraesuis (S. choleraesuis), Salmonella typhimurium (S. typhimurium), Escherichia coli (E. coli), and Brachyspira hyodysenteriae (B. hyodysenteriae). IFA and IHC results indicated that the monoclonal antibodies can be successfully used as primary antibodies to detect L. intracellularis in infected cells and in the crypt of the ileum from infected tissues of PE. Our findings suggested that the new monoclonal antibody specific against L. intracellularis will be useful for the evaluation of L. intracellularis infection in vivo and in vitro.

Evaluation of immune efficacy of Omp2 protein against Lawsonia intracellularis in mice.

Porcine proliferative enteropathy (PPE) is caused by the obligate intracellular bacterium Lawsonia intracellularis. Infection results in an enteric disease characterised by decreased growth performance of pigs, and presents a major economic burden for swine industries worldwide. Since vaccination is an effective technique for controlling PPE, novel effective vaccine platforms are need to be developed. In this study, five proteins of L. intracellularis were screened through animal experiments and the highly immunoprotective Omp2 protein was identified. Then, the immune efficacy of Omp2 was further evaluated based on humoral and cell mediated immune (CMI) responses, faecal bacterial shedding, histopathological lesions, immune barrier function of intestinal mucosa as well as digestive and absorptive capacity following challenge of mice with L. intracellularis. Mice immunised with Omp2 had reduced faecal shedding, fewer histopathological lesions and reduced bacteria colonisation of the ileum. Additionally, Omp2 immunised mice showed stronger serum IgG and IFN-γ levels, up-regulated Occludin and zonula occludens-1 (ZO-1) mRNA levels, as well as increased numbers of intestinal intraepithelial lymphocytes (IELs) and levels of sIgA. On the contrary, the activities of LPS, α-AMS and AKP were significantly increased. Our investigation indicated that immunization with Omp2 reduced the severity of clinical signs and provided efficacious immunoprotection for target animals against L. intracellularis infection in mouse model.

Dietary Eucommia ulmoides Extract Alleviates the Effect of Cold Stress on Chick Growth Performance, Antioxidant and Immune Ability.

This study aimed to investigate the protective value of Eucommia ulmoides extract (EUE) on chicks under cold stress. A total of 21 compounds were identified in EUE using mass spectrometry (LC-MS). Ninety chicks were divided into a control group (CS) fed a basal diet and an experimental group supplemented with EUE, exposed to 10 ± 1 °C for 8 h per day. Results showed, compared with the CS group, the body weights (BW) (p < 0.01) and average daily gains ADG (p < 0.05) of the EUE group were increased throughout the study period. Chicks fed EUE had higher AFI (0-7 d, p < 0.001) and lower feed-to-gain ratios (F/G) (0-15 d, p < 0.001). EUE increased the activities of superoxide dismutase (SOD) (15 d, p < 0.05) and glutathione peroxidase (GSH-Px) (7 d, p < 0.05), whereas it decreased malondialdehyde (MDA) (15 d, p < 0.01). The contents of IgA (7 d, p < 0.05), IgG (7 d; 15 d, p < 0.01), and IgM (15 d, p < 0. 001) were higher in the EUE group. Dietary EUE could also reduce chick organ damage. Overall, EUE as a natural feed additive can improve the growth performance, antioxidant capacity, and immune level, and reduce the organ damage of cold-stressed chicks.

Coinfection with porcine circovirus type 2 and Streptococcus suis serotype 2 enhances pathogenicity by dysregulation of the immune responses in piglets.

Porcine circovirus type 2 (PCV-2) and Streptococcus suis (S. suis) are common pathogens in pigs. Both pathogens are associated with the porcine respiratory disease complex. Clinically, coinfection of PCV-2 and S. suis are often detected in pigs with respiratory symptoms, while interactions between the two pathogens during coinfection and the coinfection pathogenesis are poorly understood. In this study, a piglet model coinfected with PCV-2 and Streptococcus suis serotype 2 (SS2) was established; coinfection of piglets increased the contents of SS2 in blood, and piglets showed more severe pneumonia, myocarditis and arthritis. Peripheral blood mononuclear cells (PBMCs) were collected and coinfected piglets showed high expression levels of inflammatory cytokines and TLR2, TLR4, while levels of CD4, CD8 and MHC II were reduced. In addition, in order to further explore the mechanisms of coinfection induced cytokine overexpression, an in vitro model of coinfection with PCV-2 and SS2 was established using cells of the porcine monocytic line 3D4/21. Similar to the in vivo results,coinfected cells exhibited increased expression of the cytokines IL-6, IL-8, TNF-α and the receptors TLR2, TLR4, while they showed a lower expression of MHC II than cells infected with SS2 alone. Furthermore, in coinfected 3D4/21 cells, both MAPK and NF-κB signaling pathways were activated, and the increased expression of IL-8 was related to TLR4. In general, coinfection with PCV-2 and SS2 exacerbated the inflammatory response and probably impaired macrophage antigen presentation, resulting in immune dysregulation and increasing the severity of host infection.