[Immunoadjuvant Effect of Chitosan Oligosaccharide and Its Feasibility of Being Used as an Adjuvant for Attenuated Live Bacteria Vector Vaccines]. / å£³å¯¡ç³–çš„å ç–«ä½å‰‚æ•ˆæžœåŠä½œä¸ºå‡æ¯’æ´»èŒè½½ä½“ç–«è‹—ä½å‰‚çš„å¯è¡Œæ€§æŽ¢ç´¢.

Objective: To study the immunoadjuvant effects of chitosan oligosaccharide (COS), including the immune activation and the triggering of lysosomal escape, and to explore whether COS can be used as an adjuvant for attenuated live bacteria vector vaccines. Methods: 1) Mouse macrophages RAW264.7 cells were cultured with COS at 0 mg/mL (the control group) and 0.1-4 mg/mL for 24 h and the effect on cell viability was measured by CCK8 assay. Mouse macrophages RAW264.7 were treated with COS at 0 (the control group), 1, 2, and 4 mg/mL for 24 h. Then, the mRNA expression levels of the cytokines, including IFN-γ, IL-10, TGF-ß, and TLR4, were determined by RT-qPCR assay. 2) RAW264.7 cells were treated with 1 mL of PBS containing different components, including calcein at 50 µg/mL, COS at 2 mg/mL, and bafilomycin A1, an inhibitor, at 1 µmol/mL, for culturing. The cells were divided into the Calcein group, Calcein+COS group, and Calcein+COS+Bafilomycin A1 group accordingly. Laser scanning confocal microscopy was used to observe the phagocytosis and the intracellular fluorescence distribution of calcein, a fluorescent dye, in RAW264.7 cells in the presence or absence of COS intervention to determine whether COS was able to trigger lysosomal escape. 3) LM∆E6E7 and LI∆E6E7, the attenuated Listeria vector candidate therapeutic vaccines for cervical cancer, were encapsulated with COS at the mass concentrations of 0.5 mg/mL, 1 mg/mL, 2 mg/mL , 4 mg/mL, and 8 mg/mL. Then, the changes in zeta potential were measured to select the concentration of COS that successfully encapsulated the bacteria. Phagocytosis of the vaccine strains by RAW264.7 cells was measured before and after LM∆E6E7 and LI∆E6E7 were coated with COS at 2 mg/mL. Results: 1) CCK8 assays showed that, compared with the findings for the control group, the intervention of RAW264.7 cells with COS at different concentrations for 24 h was not toxic to the cells and promoted cell proliferation, with the difference being statistically significant (P<0.05). According to the RT-qPCR results, compared with those of the control group, the COS intervention up-regulated the mRNA levels of TLR4 and IFN-γ in RAW264.7 cells, while it inhibited the mRNA expression levels of TGF-ß and IL-10, with the most prominent effect being observed in the 4 mg/mL COS group (P<0.05). 2) Laser scanning confocal microscopy revealed that the amount of fluorescent dye released from lysosomes into the cells was greater in the Calcein+COS group than that in the Calcein group. In other words, a greater amount of fluorescent dye was released from lysosomes into the cells under COS intervention. Furthermore, this process could be blocked by bafilomycin A1. 3) The zeta potential results showed that COS could successfully encapsulate the surface of bacteria when its mass concentration reached 2 mg/mL. Before and after the vaccine strain was encapsulated by COS, the phagocytosis of LM∆E6E7 by RAW264.7 cells was 5.70% and 22.00%, respectively, showing statistically significant differences (P<0.05); the phagocytosis of LI∆E6E7 by RAW264.7 cells was 1.55% and 6.12%, respectively, showing statistically significant differences (P<0.05). Conclusion: COS has the effect of activating the immune response of macrophages and triggering lysosomal escape. The candidates strains of coated live attenuated bacterial vector vaccines can promote the phagocytosis of bacteria by macrophages. Further research is warranted to develop COS into an adjuvant for bacterial vector vaccine.

Identification of a novel porcine Teschovirus 2 strain as causative agent of encephalomyelitis in suckling piglets with high mortality in China.

BACKGROUND: Porcine Teschovirus (PTV), also named Teschovirus A, is prevalent in pig populations, mainly causing neurological symptoms, diarrhea, pneumonia, and reproductive failure, however the morbidity and mortality are usually low in pig farms. CASE PRESENTATION: In this study, we reported a PTV outbreak investigation in one large-scale pig farm in China with severe symptoms including diarrhea, lethargy, locomotor ataxia, nystagmus, paralysis of the hind limbs, and coma in piglets. More importantly, the mortality reached 38% in suckling pigs, which is remarkably high in PTV history. A novel PTV strain, named HeNZ1, was isolated from cerebral samples of one suckling pig and the genome sequence was obtained by NGS sequencing. Phylogenetic and evolutionary divergence analyses revealed that HeNZ1 belongs to PTV genotype 2. Surprisingly, the VP1 coding region of HeNZ1 shares the highest sequence similarity with European PTV-2 strains, instead of China domestic PTV-2 strains, implying it may not derive from China local PTV-2 strains. Multiple sequence alignment and B cell epitope prediction of PTV VP1 and VP2 protein revealed 10 B cell epitopes, 5 mutant clusters and 36 unique mutation sites, of which 19 unique mutation sites are located in B cell epitopes and exposed on the surface of VP1 or VP2, implying significant antigenic drift potential of HeNZ1. CONCLUSION: These results indicate that HeNZ1 is a highly virulent PTV-2 strain, which capable of causing severe neurological symptoms and high mortality in piglets. Bioinformatic analysis suggest that HeNZ1 is genetically and antigenically different from other Chinese PTV-2 strains. Overall, current case expanded our understanding of PTV-2 clinical spectrum and revealed the emergence of a highly virulent PTV-2 strain with substantial genetic diversity and antigenic drift potential in VP1 and VP2.

Molybdenum and cadmium co-exposure promotes M1 macrophage polarization through oxidative stress-mediated inflammatory response and induces pulmonary fibrosis in Shaoxing ducks (Anas platyrhyncha).

High molybdenum (Mo) and cadmium (Cd) are harmful to the body, but pulmonary toxicity induced by Mo and Cd co-exposure is unknown. To assess the combined impacts of Mo and Cd on fibrosis through M1 polarization in the lung of ducks, 80 healthy 8-day-old Shaoxing ducks (Anas platyrhyncha) were randomly assigned to 4 groups and fed with containing unequal doses of Mo or/and Cd diet. Lung tissues were collected on the 16th week. Results indicated that Mo or/and Cd significantly increased their contents in the lungs, and led to trace elements disorder and histological abnormality, and oxidative stress accompanied by promoting contents of H2 O2 and MDA and decreasing activities of T-SOD, GSH-Px, and CAT, then activated the TLR4/NF-κB/NLRP3 pathway accompanied by upregulating Caspase-1, ASC, IL-18, IL-1ß, TLR4, NF-κB, and NLRP3 expression levels, and disrupted M1/M2 balance to divert toward M1, which evoked the TGF-ß/Smad2/3-mediated fibrosis by elevating TGF-ß1, Smad2, Smad3, COL1A1, α-SMA, and MMP2 expression levels, and decreasing Smad7 and TIMP2 expression levels. The changes of the combined group were most obvious. To sum up, the research demonstrated that Mo or/and Cd may cause macrophages to polarize toward M1 by oxidative stress-mediated the TLR4/NF-κB/NLRP3 pathway, then result in fibrosis through the TGF-ß1/Smad2/3 pathway in duck lungs. Mo and Cd may worsen lung damage.

Molybdenum and cadmium co-induce hypothalamus toxicity in ducks via disturbing Nrf2-mediated defense response and triggering mitophagy.

Growing evidences reveal that Nrf2-mediated antioxidant defense response and mitophagy are involved in the toxic mechanism of heavy metals, but the effects of molybdenum (Mo) and cadmium (Cd) co-exposure on Nrf2-mediated antioxidant defense response and mitophagy in duck hypothalamus have yet to be elucidated. Herein, 40 healthy 7-day-old ducks were randomly assigned to 4 groups and fed diets containing different doses of Mo or/and Cd for 16 weeks, respectively. The data demonstrated that Mo or/and Cd notably elevated their contents in hypothalamus, decreased Cu, Fe, Zn and Se contents, caused pathological damage and oxidative stress accompanied by elevating MDA content and reducing CAT, T-AOC, T-SOD, GSH-Px activities. Moreover, Mo or/and Cd not only restrained Nrf2 pathway by decreasing Nrf2, HO-1, NQO1, GST, CAT, SOD1, GCLM mRNA expression levels and Nrf2 protein expression level, but also disturbed mitochondrial dynamics and triggered PINK1/Parkin-mediated mitophagy by enhancing MFF, PINK1, Parkin, Bnip3, LC3A, LC3B mRNA expression levels and PINK1, Parkin, LC3B-II/LC3B-I protein expression levels, inhibiting Mfn1, Mfn2, OPA1, P62 mRNA expression levels and P62 protein expression level, and facilitating the colocalization between LC3 and HSP60. The changes of above factors were most remarkable under Mo and Cd co-treatment. Overall, the results elucidate that Mo and Cd can synergistically inhibit Nrf2-mediated antioxidant defense response and activate PINK1/Parkin pathway-dependent mitophagy in duck hypothalamus, whose mechanism is somehow related to Mo and Cd accumulation.

The synergistic effect of traditional Chinese medicine prescription and rumen-protected γ-aminobutyric acid on beef cattle under heat stress.

Traditional Chinese medicine (TCM) prescription or rumen-protected GABA (RP-GABA) can effectively relieve the heat stress (HS) in cattle, but the joint effects of TCM and RP-GABA on HS in beef cattle are not fully clarified. To investigate the effects of TCM or/and RP-GABA on growth performance, antioxidant capacity, serum parameters and heat shock proteins (HSPs) expression in beef cattle under HS ambient. A total of 40 Jinjiang yellow cattle were randomly divided into four groups: (a) control group (basal diet, BD), (b) TCM group (BD+TCM), (c) GABA group (BD+RP-GABA) and (d) TCM+GABA group (BD+TCM plus RP-GABA). Results indicated that the average daily feed intake (ADFI) was significantly elevated in the TCM+GABA group (p < 0.05), whereas, average daily gain (ADG) was elevated (p < 0.05) in the group of TCM (38.5%), GABA (35.4%) and TCM+GABA (41.5%) compared with the control group. Meanwhile, TCM+GABA exhibited prominently more positive effects in terms of SOD, BUN, T-CHO, TG, HDL-C and HSP70 (p < 0.05 or p < 0.01) than the control and other treatment groups. Therefore, TCM or GABA can effectively moderate the HS response in beef cattle by ameliorating antioxidant capacity, serum parameters and HSPs expression, meanwhile, the combination of them exerts a synergistic effect on HS alleviation.

Effects of molybdenum and cadmium on the oxidative damage and kidney apoptosis in Duck.

Molybdenum (Mo) is an essential element for human beings and animals; however, high dietary intake of Mo can lead to adverse reactions. Cadmium (Cd) is one of the major transitional metals which has toxic effects in animals. To investigate the co-induced toxic effects of Mo and Cd on oxidative damage and kidney apoptosis in duck, 120 ducks were randomly divided into control group and 5 treatment groups which were treated with a commercial diet containing different dosages of Mo and Cd. Kidney samples were collected on the 60th and 120th days to determine the mRNA expression levels of ceruloplasmin (CP), metallothionein (MT), Bak-1, and Caspase-3 by quantitative RT-PCR. Additionally, we also determined the antioxidant activity indexes and contents of Mo, Cd, copper (Cu), iron (Fe), zinc (Zn), and selenium (Se) in serum. Meanwhile, ultrastructural changes of the kidney were observed. The results showed that glutathione reductase (GR) activity and CP level in serum were decreased in combination groups. In addition, the antioxidant indexes were decreased in co-treated groups compared with single treated groups. The mRNA expression levels of Bak-1 and Caspase-3 increased in co-treated groups. The mRNA expression level of CP in high-dose combination group was downregulated, while the mRNA expression of MT was upregulated except for low-dose Mo group. Additionally, in the later period the content of Cu in serum decreased in joint groups while the contents of Mo and Cd increased. In addition, ultrastructural changes showed mitochondrial crest fracture, swelling, deformed nuclei, and karyopyknosis in co-treated groups. Taken together, it was suggested that dietary Mo and Cd might lead to oxidative stress, kidney apoptosis and disturb homeostasis of trace elements in duck, and it showed a possible synergistic relationship between the two elements.

Traditional chinese medicine prescriptions enhance growth performance of heat stressed beef cattle by relieving heat stress responses and increasing apparent nutrient digestibility.

The present aim was to investigate the effects of traditional Chinese medicine prescriptions (TCM) on body temperature, blood physiological parameters, nutrient apparent digestibility and growth performance of beef cattle under heat stress conditions. Twenty-seven beef cattle were randomly divided into three groups as following; i) high temperature control (HTC), ii) traditional Chinese medicine prescriptions I+high temperature (TCM I) and iii) traditional Chinese medicine prescriptions II+high temperature (TCM II) (n = 9 per group). The results showed that the mean body temperature declined in TCM II treatment (p<0.05). Serum T3 and T4 levels with TCM I and TCM II treatments elevated (p<0.05), and serum cortisol levels of TCM I treatments decreased (p<0.05), compared with the HTC group. Total protein, albumin, globulin in TCM II treatments elevated and blood urea nitrogen levels of both TCM treatments increased, but glucose levels of both TCM treatments decreased, compared with the HTC group (p<0.05). The apparent digestibility of organic matter and crude protein with TCM I treatment increased, and the apparent digestibility of acid detergent fiber elevated in both TCM treatments (p<0.05). Average daily feed intake was not different among three groups, however average daily gain increased and the feed:gain ratio decreased with both TCM treatments, compared with the HTC group (p<0.05). The present results suggest that dietary supplementation with TCM I or TCM II improves growth performance of heat stressed beef cattle by relieving heat stress responses and increasing nutrient apparent digestibility.

Molybdenum and Cadmium Co-induce Pyroptosis via Inhibiting Nrf2-Mediated Antioxidant Defense Response in the Brain of Ducks.

Excess molybdenum (Mo) and cadmium (Cd) are harmful to animals, but the neurotoxic mechanism co-induced by Mo and Cd is unclear. To estimate the effects of Mo and Cd co-exposure on pyroptosis by nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated antioxidant defense response in duck brains, 40 healthy 7-day-old ducks were randomly assigned to 4 groups and fed diet supplemented with Mo or/and Cd for 16 weeks, respectively. Results showed that Mo or/and Cd markedly increased Mo and Cd contents; decreased iron (Fe), copper (Cu), zinc (Zn), and selenium (Se) contents, elevated malondialdehyde (MDA) content; and decreased total-antioxidant capacity (T-AOC), total-superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activities accompanied by pathological damage in brain. Additionally, Mo or/and Cd inhibited Nrf2 pathway via decreasing Nrf2, CAT, SOD1, glutathione S-transferase (GST), hemeoxygenase-1 (HO-1), NAD (P) H:quinone oxidoreductase 1 (NQO1), glutamate-cysteine ligase catalytic subunit (GCLC), and modifier subunit (GCLM) mRNA levels and Nrf2 protein level, which induced pyroptosis through upregulating nucleotide oligomerization domain-like receptor protein-3 (NLRP3), apoptosis-associated speck-like protein (ASC), gasdermin A (GSDMA), gasdermin E (GSDME), interleukin-1ß (IL-1ß), interleukin-18 (IL-18), Caspase-1, NIMA-related kinase 7 (NEK7) mRNA levels and NLRP3, Caspase-1 p20, gasdermin D (GSDMD), ASC protein levels and IL-1ß, and IL-18 contents. Besides, the changes of these indicators were most apparent in the Mo and Cd co-treated group. Collectively, the results certificated that Mo and Cd might synergistically induce pyroptosis via inhibiting Nrf2-mediated antioxidant defense response in duck brains, whose mechanism is closely related to Mo and Cd accumulation.

PRRSV Elimination in a Farrow-to-Finish Pig Herd Using Herd Closure and Rollover Approach.

It is well established that PRRSV elimination is an effective strategy for PRRS control, but published reports concerning successful PRRSV elimination cases in farrow-to-finishing herds are rare. Here, we have reported a successful PRRSV elimination case in a farrow-to-finish herd by employing a "herd closure and rollover" approach with some modifications. Briefly, the introduction of pigs to the herd was stopped and normal production processes were maintained until the herd reached a PRRSV provisional negative status. During the herd closure, strict biosecurity protocols were implemented to prevent transmission between nursery pigs and sows. In the current case, introducing gilts before herd closure and live PRRSV exposure were skipped. In the 23rd week post-outbreak, the pre-weaning piglets started to show 100% PRRSV negativity in qPCR tests. In the 27th week, nursery and fattening barns fully launched depopulation. In the 28th week, nursery and fattening houses reopened and sentinel gilts were introduced into gestation barns. Sixty days post-sentinel gilt introduction, the sentinel pigs maintained being PRRSV antibody negative, manifesting that the herd matched the standard of the provisional negative status. The production performance of the herd took 5 months to bounce back to normal. Overall, the current study provided additional information for PRRSV elimination in farrow-to-finish pig herds.

Cadmium and molybdenum co-induce pyroptosis and apoptosis via the PTEN/PI3K/AKT axis in the livers of Shaoxing ducks (Anas platyrhynchos).

Cadmium (Cd) and excessive molybdenum (Mo) have adverse impacts on animals. However, the hepatotoxicity co-induced by Cd and Mo in ducks has not been fully elucidated. In order to explore the impacts of Cd and Mo co-exposure on pyroptosis and apoptosis by the PTEN/PI3K/AKT pathway in the livers of ducks, 40 healthy 7-day-old Shaoxing ducks (Anas platyrhynchos) were randomly assigned into 4 groups, and Cd or/and Mo were added to the basic diet per kilogram (kg): control group (0 mg Mo and 0 mg Cd), Mo group (100 mg Mo), Cd group (4 mg Cd), and Mo + Cd group (100 mg Mo and 4 mg Cd), with 16 weeks feed management. Results signified that Cd or/and Mo caused trace element imbalance, liver function and histomorphological abnormalities in the duck liver, and activated the PTEN/PI3K/AKT pathway through increasing PTEN mRNA and protein levels, reducing PI3K, AKT mRNA and p-AKT/AKT protein levels, which triggered pyroptosis and apoptosis via increasing Caspase-1, NLRP3, NEK7, ASC, GSDME, GSDMA, IL-1ß and IL-18 mRNA levels, Caspase-1 p20, NLRP3, ASC and GSDMD protein levels, and IL-1ß and IL-18 contents, and increasing Bak-1, Bax, Cyt C and Caspase-3 mRNA levels and cleaved Caspase-3/Caspase-3 protein level, and downregulating Bcl-2 mRNA level and the ratio of Bcl-2 to Bax, respectively. Overall, the results illustrate that pyroptosis and apoptosis induced by Cd or/and Mo may be associated with activating the PTEN/PI3K/AKT pathway in the livers of ducks. There may be a synergy between these two elements.

Involvement of calcium homeostasis and unfolded protein response in autophagy co-induced by molybdenum and cadmium in duck (Anas platyrhyncha) brain.

Excess molybdenum (Mo) and cadmium (Cd) are harmful to animals, but neurotoxicity caused by Mo and Cd co-exposure in ducks is yet unknown. To assess joint impacts of Mo and Cd on autophagy via calcium homeostasis and unfolded protein response (UPR) in duck brain, 40 healthy 7-day-old ducks (Anas platyrhyncha) were assigned to 4 groups at random and fed diets supplemented with different doses of Mo or/and Cd for 16 weeks, respectively. Brain tissues were excised for experiment. Results exhibited that Mo or/and Cd disturbed calcium homeostasis by decreased ATPase activities and increased calcium (Ca) content, and upregulated calcium homeostasis-related factors Ca2+/CAM-dependent kinase IIɑ (CaMKIIɑ), calcineurin (CaN), inositol-1,4,5-trisphosphate receptor (IP3R), and calreticulin (CRT) expression levels. Meanwhile, the upregulation of UPR-related factor expression levels indicated that Mo or/and Cd activated UPR. Moreover, Mo or/and Cd triggered autophagy through promoting the number of autophagosomes and LC3II immunofluorescence intensity and altering autophagy key factor expression levels. Correlation analysis showed that there were obvious connections among Ca2+ homeostasis, endoplasmic reticulum (ER) stress, and autophagy induced by Mo or/and Cd. Thence, it can be speculated that autophagy initiated by Mo or/and Cd may be associated with interfering Ca2+ homeostasis and triggering UPR.

Phenylalanine-arginine ß-naphthylamide could enhance neomycin-sensitivity on Riemerella anatipestifer in vitro and in vivo.

Riemerella anatipestifer is an important duck pathogen responsible for septicemia and infectious serositis, which has caused great economic losses to the duck industry. Phenylalanine-arginine ß-naphthylamide (PAßN) is an efflux pump inhibitor, which mainly reduces the efflux effect by competing with antibiotics for efflux pump channels. Here, we found that R. anatipestifer strain GD2019 showed resistances to gentamicin, amikacin, kanamycin, and neomycin. Notably, PAßN could significantly reduce the Minimal inhibitory concentrations (MICs) of neomycin on the GD2019 strain. Moreover, PAßN combined with neomycin significantly decreased bacterial loads, relieved pathological injury and increase survival rate (p < 0.05) for the ducks lethally challenged by the GD2019 strain. Therefore, our results suggested, in vitro and in vivo, PAßN could reduce neomycin-resistant of R. anatipestifer. Importantly, finding of this study provide a new approach for treating antibiotic-resistant R. anatipestifer infection.

Baicalin ameliorates APEC-induced intestinal injury in chicks by inhibiting the PI3K/AKT-mediated NF-κB signaling pathway.

Avian pathogenic Escherichia coli (APEC) is the causative agent of avian colibacillosis. Baicalin (BA) possesses multiple pharmacological effects, but the mechanism underlying its activity in APEC-induced intestinal injury remains unknown. This study aims to investigate the protective effects and possible mechanism of BA against APEC-induced intestinal injury. Sixty 1-day-old chicks were randomly divided into 4 groups: the control group (basal diet), E. coli group (basal diet), BAI10 group (10 mg/kg BA), and BAI20 group (20 mg/kg BA). After pretreatment with BA for 15 d and subsequent induction of APEC infection by pectoralis injection, the ileum was collected and analyzed. The results showed that BA-pretreatment demonstrated an alleviation of chicks in diarrhea rate, mortality, and histopathological changes in intestinal tissues after APEC infection. Additionally, following APEC infection, BA improved the intestinal barrier by elevating zona occludens (ZO)s (ZO-1, 2, 3), Claudins (Claudin1, 2, 3), Occludin, avian ß-defensin (AvBD)s (AvBD1, 2, 4), lysozyme (Lyz) mRNA levels and ZO-1, Claudin1, and Occludin protein levels. Besides, the activities of total superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) and the SOD-1 and CAT mRNA levels and SOD-1 protein level were elevated by BA pretreatment. BA pretreatment also decreased the malondialdehyde (MDA) content, heme oxygenase-1 (HO-1) and NADH quinone oxidoreductase 1 (NQO1) mRNA levels, and HO-1 protein level after APEC infection. BA alleviated the APEC-induced inflammatory response, including downregulating the mRNA levels of proinflammatory cytokines (tumor necrosis factor-α (TNF-α), interleukin [IL]-1ß, IL-6, IL-8) and upregulating the mRNA levels of anti-inflammatory cytokines (IL-4, IL-10, IL-13, transforming growth factor-ß [TGF-ß]). Furthermore, BA decreased the mRNA and protein levels of phosphatidylinositol 3 kinase (PI3K), protein kinase B (AKT), and nuclear factor kappa-B (NF-κB) as well as the expression of the phosphorylated forms of these proteins after APEC infection. Collectively, our findings indicate that BA exerts a protective effect against APEC-induced intestinal injury in chicks by inhibiting the PI3K/AKT-mediated NF-κB pathway, suggesting that BA may be a potential therapeutic approach for avian colibacillosis.

Luteolin Attenuates APEC-Induced Oxidative Stress and Inflammation via Inhibiting the HMGB1/TLR4/NF-κB Signal Axis in the Ileum of Chicks.

Avian pathogenic E. coli (APEC) is typically the cause of avian colibacillosis, which can result in oxidative stress, inflammation, and intestinal damage (APEC). Luteolin, in the form of glycosylation flavone, has potent anti-inflammatory and anti-oxidative properties. However, its effects on APEC-induced intestinal oxidative stress and NF-κB-mediated inflammation in chicks remains poorly understood. After hatching, one-day-old chicks were stochastically assigned to four groups: a control group (basic diet), an E. coli group (basic diet) and L10 and L20 groups (with a dry matter of luteolin diet 10 mg/kg and 20 mg/kg, respectively), with fifteen chicks in each group and one repeat per group. They were pretreated for thirteen days. The body weight, mortality, histopathological changes in the ileum, antioxidant status, and the mRNA and protein-expression levels of factors associated with the HMGB1/TLR4/NF-κB signal axis of the chicks were measured. The results showed that luteolin treatment decreased the mRNA and protein-expression level of the related factors of HMGB1/TLR4/NF-κB signal axis in the ileum, reduced inflammation, increased antioxidant enzyme activity, and reduced intestinal injury. Collectively, luteolin alleviated APEC-induced intestinal damage by means of hindering the HMGB1/TLR4/NF-κB signal axis, which suggests that luteolin could be a good method for the prevention and treatment of avian colibacillosis.

Nephropathogenic Infectious Bronchitis Virus Mediates Kidney Injury in Chickens via the TLR7/NF-κB Signaling Axis.

Nephropathogenic infectious bronchitis virus (NIBV) is one of the most important viral pathogens in the world poultry industry. Here, we used RT-qPCR, WB and immunofluorescence to explore the interaction between NIBV and the host innate immune system of the kidney. Multiple virions were found in the kidney tissues of the disease group under electron microscopy, and pathological changes such as structural damage of renal tubules and bleeding were observed by HE staining. In addition, we found that the mRNA levels of TLR7, TRAF6, and IKKß were upregulated after NIBV infection. IRF7 mRNA levels decreased significantly at 5 dpi and increased significantly at 11 to 18 dpi. The NF-κB P65 mRNA level increased significantly at 5 to 18 dpi and decreased at 28 dpi. However, NIBV infection-induced NF-κB P65 protein levels were downregulated at multiple time points. Moreover, we demonstrated that the cytokine (IFN-γ, IL-8, and IL-6) mRNA and protein expression levels were increased significantly at multiple time points after NIBV infection. Furthermore, immunofluorescence analysis showed that NF-κB P65 and IFN-γ were mainly located in the nuclear or perinuclear region. The positive signal intensity of NF-κB P65 was significantly lower than that of the normal group at 1 to 5 dpi, and there was no significant change in the subsequent time period. The positive signal intensity of IFN-γ decreased significantly at 5 dpi, and increased significantly at 11 to 28 dpi. In conclusion, we found that NIBV promoted cytokine release through the TLR7/NF-κB signaling axis, thus causing kidney injury.

Response of Growth Performance, Blood Biochemistry Indices, and Rumen Bacterial Diversity in Lambs to Diets Containing Supplemental Probiotics and Chinese Medicine Polysaccharides.

This study aims to investigate the effects of probiotics and Chinese medicine polysaccharides (CMPs) on growth performance, blood indices, rumen fermentation, and bacteria composition in lambs. Forty female lambs were randomly divided into four groups as follows: control, probiotics, CMP, and compound (probiotics + CMP) groups. The results showed that probiotics treatment increased the concentrations of blood glucose (GLU) and immunoglobulin G (IgG) and enhanced rumen microbial protein contents but declined the value of pH in rumen fluid compared with the control (P < 0.05). Furthermore, supplementation with CMP enhanced the average daily gain (ADG) and the contents of IgA, IgG, and IgM in the serum but decreased the F:G ratio compared with the control (P < 0.05). Besides, both CMP and compound (probiotics + CMP) treatments decreased the ratio of acetic acid and propionic acid compared with the control (P < 0.05). High-throughput sequencing data showed that at the genus level, the relative abundance of Veillonellaceae_UCG-001 in the probiotics group was increased, the relative abundance of Succiniclasticum and norank_f__Muribaculaceae in the CMP group were enhanced, and the relative abundance of Ruminococcaceae_UCG-002 in the compound group was raised compared with the control (P < 0.05). In summary, supplementation with probiotics can promote rumen protein fermentation but decrease the diversity of bacteria in rumen fluid; however, CMP treatment increased the relative abundance of Fibrobacteria, changed rumen microbial fermentation mode, increased the immune function, and ultimately improved the growth performance.

The protection of selenium against cadmium-induced mitophagy via modulating nuclear xenobiotic receptors response and oxidative stress in the liver of rabbits.

Cadmium (Cd) is a harmful heavy metal that can cause many health problems, while selenium (Se) is an essential nutrient for organisms that can protect them from heavy metal-induced damage. To explore the effects of Se on Cd-induced mitophagy in the liver, forty 3-month-old New Zealand white rabbits (2-2.5 kg), half male and half female, were randomly divided into four groups: the Control group, the Se (0.5 mg/kg body weight (BW)) group, the Cd (1 mg/kg BW) group and the Se+Cd group. After 30 days, the toxicity from Cd in the liver was assessed in terms of the nuclear xenobiotic receptor (NXR) response, oxidative stress and mitophagy. It was found that Cd decreased the activities of CYP450 enzymes and antioxidant enzymes and increased the contents of malondialdehyde (MDA) and hydrogen peroxide (H2O2) and also increased the consumption of reduced glutathione (GSH). Moreover, the mRNA levels of NXRs (CAR, PXR, AHR and Nrf2), some mitochondrial function factors (PGC-1α, Sirt1, Sirt3, Nrf1 and TFAM) and mitochondrial fusion factors (Mfn1, Mfn2 and OPA1) were downregulated, but the mRNA levels of other mitochondrial function factors (VDAC1, Cyt C and PRDX3), mitochondrial fission factors (Fis1 and MFF) and those in the PINK1/Parkin-mediated mitophagy pathway (p62, Bnip3 and LC3) were upregulated under Cd exposure. The protein expression levels of Nrf2, SOD2, PGC-1α, PINK1 and Parkin were consistent with the mRNA expression levels in the Cd group. Se alleviated the changes in the abovementioned factors induced by Cd. In conclusion, the results indicate that Cd can cause oxidative stress in rabbit livers by inhibiting NXRs and the antioxidation response leading to mitophagy, and these harmful changes caused by Cd can be alleviated by Se.

Puerarin improved growth performance and postmortem meat quality by regulating lipid metabolism of cattle under hot environment.

This study aims to evaluate the effect of puerarin on performance, meat quality, and serum indexes of beef cattle under hot environment. Thirty-two bulls were divided into four groups and fed diet supplemented with puerarin at 0, 200, 400, or 800 mg/kg. Results showed that heat stress was employed for 54 out of 60 days, 400 mg/kg group declined serum cortisol (COR) contents, all treatments increased the contents of total cholesterol, high density lipoprotein cholesterol, and total superoxide dismutase activity; in addition, glutathione peroxidase activity of 200 mg/kg group were enhanced, only 800 mg/kg group enhanced immunoglobulin (IgA, IgM, and IgG) and low density lipoprotein cholesterol contents compared with the control (p < .05). Moreover, 400-mg/kg puerarin increased serum growth hormone levels compared with 200 mg/kg group but declined COR concentrations compared with 200 mg/kg and 800 mg/kg groups (p < .05). More importantly, average daily gain and daily matter intake, and intramuscular fat contents of 400 mg/kg group were enhanced, but the shear force of beef in 400 mg/kg and 800 mg/kg groups were declined compared with the control (p < .05). These findings indicated that supplemental with puerarin enhanced immune and antioxidant, and 400 mg/kg of puerarin improved performance and meat quality by normalizing levels of stress hormones and increasing intramuscular fat deposition of beef cattle under hot environment.

Traditional Chinese Medicine Prescriptions Decrease Diarrhea Rate by Relieving Colonic Inflammation and Ameliorating Caecum Microbiota in Piglets.

Diarrhea is a leading cause of death in piglets. XiaoJianZhong (XJZ) and Jingsananli-sepsis (JSS) were two traditional Chinese medicine (TCM) prescriptions to prevent and treat intestinal diseases, including diarrhea and inflammatory disease. Here, we investigated the effects of XJZ and JSS on diarrhea rate, growth performance, colonic inflammation, and caecum microbiota in piglets. A total of 18 piglets were selected and randomly divided into three groups. Control group was supplied with basal diets, while TCM1 and TCM2 groups were, respectively, supplied with XJZ and JSS in basal diets. Decreased diarrhea rate, colonic or caecal pH, and elevated apparent nutrient digestibility were observed in both TCM groups. Meanwhile, both prescriptions alleviated colonic inflammation by decreasing mRNA expression of proinflammatory cytokines and suppressing the TLR4/MyD88/NF-κB signaling pathway. Additionally, TCM1 and TCM2 prescriptions ameliorated caecum microbiota composition and increased the abundance of beneficial bacteria, together with regulations on several genes that are responsible for signaling pathways involved in cancers and metabolic diseases. Importantly, both TCM1 and TCM2 significantly promoted the average daily gain (ADG) and reduced the feed : gain (F : G) ratio. In conclusion, both TCM prescriptions effectively decreased diarrhea rate and increased growth performance by elevating apparent nutrient digestibility and gut health, via relieving colonic inflammation and ameliorating gut microbiota composition of piglets.

The anti-heat stress effects of Chinese herbal medicine prescriptions and rumen-protected γ-aminobutyric acid on growth performance, apparent nutrient digestibility, and health status in beef cattle.

This study aims to evaluate the anti-heat stress effect of Chinese herbal medicine (CHM) prescription, rumen-protected γ-aminobutyric acid (RP-GABA), and CHM plus RP-GABA co-medication on growth performance, apparent digestibility, and serum parameters in heat-stressed beef cattle. Forty beef cattle were randomly divided into four groups. Control group was supplied with basal diet, while CHM, γ-aminobutyric acid (GABA), and CHM + GABA groups were, respectively, supplied with CHM, RP-GABA, and CHM plus RP-GABA in basal diet. Our result indicated that CHM + GABA elevated apparent digestibility including crude protein, Ca, P, crude fat (CF) (p < .01), and neutral detergent fiber (NDF) (p < .05), but no difference was found with CF and NDF digestibility both in CHM and GABA group (p > .05). More importantly, average daily gain (ADG) was improved in CHM, GABA, and CHM + GABA groups, while average daily feed intake (ADFI) significantly increased only in CHM + GABA groups (p < .05). Meanwhile, CHM + GABA displayed notably more positive effect in serum hormones, immune globulin, ions contents, and blood inflammatory cytokines than other treatment groups and control group. These results demonstrated that both CHM and GABA are effective in alleviating heat stress response and the co-medication has a synergistic effect on anti-heat stress.