Mycoplasma gallisepticum (MG) is an important avian pathogen that causes significant economic losses in the poultry industry. Surprisingly, the limited protection and adverse reactions caused by the vaccines, including live vaccines, bacterin-based (killed) vaccines, and recombinant viral vaccines is still a major concern. Mycoplasma gallisepticum strains vary in infectivity and virulence and infection may sometimes unapparent and goes undetected. Although extensive research has been carried out on the biology of this pathogen, information is lacking about the type of immune response that confers protection and selection of appropriate protective antigens and adjuvants. Regardless of numerous efforts focused on the development of safe and effective vaccine for the control of MG, the use of modern DNA vaccine technology selected in silico approaches for the use of conserved recombinant proteins may be a better choice for the preparation of novel effective vaccines. More research is needed to characterize and elucidate MG products modulating MG-host interactions. These products could be used as a reference for the preparation and development of vaccines to control MG infections in poultry flocks.

The thymus is a primary lymphoid organ and plays a critical role in the immune response against infectious agents. Baicalin is a naturally derived flavonoid famous for its pharmacological properties, but the preventive effects of baicalin against immune impairment remain unclear. We examined this effect in the context ofMycoplasma gallisepticum(MG) infection-induced structural damage in the chicken thymus. Histopathological examination showed that the compact arrangement of cells in the thymus was lost in the MG-infected group. Inflammatory cell infiltration and nuclear debris accumulated, and the boundary between the cortex and medulla was not clearly visible. The mRNA and protein expression of apoptosis-related genes were significantly increased in the MG-infected group compared to the control group and the baicalin group. The number of positively stained nuclei in the terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay were increased in the MG-infected group. In addition, electron microscopic examination showed chromatin condensation, mitochondrial swelling and apoptotic vesicles in the MG-infected group. However, baicalin treatment significantly alleviated the oxidative stress and apoptosis induced by MG infection. Importantly, the abnormal morphology was partially ameliorated by baicalin treatment. Compared to the MG-infected group, the baicalin-treated group showed significantly reduced expression of apoptosis-related genes at both the mRNA and protein levels. Meanwhile, the nuclear factor erythroid 2-related factor 2 (Nrf2) signalling pathway and downstream genes were significantly upregulated by baicalin to counteract MG-induced oxidative stress and apoptosis in the thymocytes of chickens. In summary, these findings suggest that baicalin treatment efficiently attenuated oxidative stress and apoptosis by activating the Nrf2 signalling pathway and could protect the thymus from MG infection-mediated structural and functional damage.

Previous studies reported that Mycoplasma gallisepticum (MG) causes immune dysregulation in chickens. However, the underlying mechanisms of immune dysregulation in chickens are still unclear. The thymus is a primary lymphoid organ where the proliferation, differentiation and selection of T-lymphocytes occur, whereas T-lymphocytes play a crucial role in innate immune responses. To evaluate the effects of MG-infection on chicken thymus, White Leghorn chickens were divided into (1) control group and (2) MG-infection group. ATPase activities were detected by commercial kits. The hallmarks of inflammation, autophagy and energy metabolism were examined in chicken thymus tissues by histopathology, transmission electron microscopy, immunofluorescence microscopy, RT-PCR and western blotting. Immunofluorescence examination revealed that the number of CD8⁺ lymphocytes has significantly reduced in MG-infection group. In addition, morphological analysis revealed that MG induced inflammatory cells infiltration. The mitochondria were swollen and chromatin material was condensed in MG-infection group. The mRNA and protein expression results showed that MG-infection triggered the nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 3 (NLRP3) inflammasome through TLR-2/MyD88/NF-κB signaling pathway. Meanwhile, the expressions of autophagy-related genes were reduced both at mRNA and protein level in MG-infection group. While, ATPase activities and the expression of energy metabolism-related genes were reduced in the thymus of MG-infected chickens. These results showed that MG-infection triggered inflammatory response through TLR-2/MyD88/NF-κB signaling pathway, activated NLRP3 inflammasome, reduced the level of autophagy and impaired energy metabolism, which then lead to tissue damage in chicken thymus. The data provide new insights in MG-infection-mediated immune damage and provide possible therapeutic targets for future targeted therapy.

Reversed micelles were used to extract lectin from red kidney beans and factors affecting reverse micellar systems (pH value, ionic strength and extraction time) were studied. The optimal conditions were extraction at pH 4–6, back extraction at pH 9–11, ion strength at 0.15 M NaCl, extraction for 4–6 minutes and back extraction for 8 minutes. The reverse micellar system was compared with traditional extraction methods and demonstrated to be a time-saving method for the extraction of red kidney bean lectin. Mitogenic activity of the lectin was reasonably good compared with commercial phytohemagglutinin (extracted fromPhaseolus vulgaris) Mitogenic properties of the lectin were enhanced when four Chinese herbal polysaccharides were applied concurrently, among which 50 μg/mLAstragalus mongholicuspolysaccharides (APS) with 12.5 μg/mL red kidney bean lectin yielded the highest mitogenic activity and 100 mg/kg/bw APS with 12.5 mg/kg/bw red kidney bean lectin elevated mouse nonspecific immunity.

In this study, a simultaneous water extraction process for baicalin, wogonoside, and chlorogenic acid has been optimized. The effect of extraction temperature, extraction time, and liquid–solid ratio was scrutinized by single factor experiments and further analyzed by Box–Behnken design (BBD) approach using response surface methodology (RSM). The extraction yield of investigated compounds was determined by high performance liquid chromatography (HPLC). Single-factor experiments and response surface analysis results revealed that the optimized conditions are: Liquid to solid ratio 25:1 (mL/g), extraction temperature 93 °C, extraction time 2.4 h, and the extraction cycle two. Importantly, it has been noted that under the above conditions, concentrations of baicalin, wogonoside, and chlorogenic were 0.078, 0.031, and 0.013 mg/mL, respectively, and the overall desirability (OD) value was 0.76 which was higher than the non-optimized conditions and the deviation from the predicted OD value was only 2.44%. Conclusively, it has been suggested that the model was stable and feasible, and fit for extraction of baicalin, wogonoside, and chlorogenic acid from Scutellariae Radix and Lonicerae (L.) japonicae Flos.