Clostridium perfringens α-toxin is one of the major virulence factors during C. perfringens infection, causing hemolysis of erythrocytes in various species. Here, genetically engineered Lactobacillus casei (pPG-α/L. casei 393) constitutively expressing the toxoid of C. perfringens α-toxin was generated and its immunogenicity in mice for induction of protective immunity against the αtoxin was evaluated via oral immunization. The α-toxoid was constitutively expressed by pPG-α/ L. casei 393 without a specific inducer, as confirmed by western blotting, laser confocal microscopy, and flow cytometry. In an experiment on BALB/c mice to evaluate the oral immunogenicity of pPG-α/L. casei 393, significant levels of a specific secretory IgA (sIgA) antibody in the intestinal mucus and feces and an IgG antibody in the serum of the probiotic vaccine group were detected after booster immunization (p < 0.05) as compared with the pPG/L. casei 393 and PBS control groups. These antibodies effectively neutralized C. perfringens natural α-toxin. Moreover, significantly higher levels of cytokines IL-2, IL-4, IL-10, IL-12, IL-17, and interferon (IFN) γ in the serum and increased proliferation of spleen lymphocytes obtained from mice orally immunized with pPG-α/L. casei 393 were detected. With a commercial C. perfringens type A inactivated vaccine as a control, immune protection provided by the probiotic vaccine against C. perfringens α-toxin was evaluated, and 90% and 80% protection rates were observed, respectively. Therefore, strain pPGα/L. casei 393 effectively elicited mucosal, humoral, and cellular immunity, suggesting that pPG-α/ L. casei 393 is a promising candidate for development of a vaccine against C. perfringens α-toxin.
In this study, a multicenter evaluation of the Life Technologies TaqMan(®) Array Card (TAC) with 21 custom viral and bacterial respiratory assays was performed on the Applied Biosystems ViiA™ 7 Real-Time PCR System. The goal of the study was to demonstrate the analytical performance of this platform when compared to identical individual pathogen specific laboratory developed tests (LDTs) designed at the Centers for Disease Control and Prevention (CDC), equivalent LDTs provided by state public health laboratories, or to three different commercial multi-respiratory panels. CDC and Association of Public Health Laboratories (APHL) LDTs had similar analytical sensitivities for viral pathogens, while several of the bacterial pathogen APHL LDTs demonstrated sensitivities one log higher than the corresponding CDC LDT. When compared to CDC LDTs, TAC assays were generally one to two logs less sensitive depending on the site performing the analysis. Finally, TAC assays were generally more sensitive than their counterparts in three different commercial multi-respiratory panels. TAC technology allows users to spot customized assays and design TAC layout, simplify assay setup, conserve specimen, dramatically reduce contamination potential, and as demonstrated in this study, analyze multiple samples in parallel with good reproducibility between instruments and operators.
Type II transmembrane serine proteases (TTSPs) facilitate the spread and replication of viruses such as influenza and human coronaviruses, although it remains unclear whether TTSPs play a role in the progression of animal coronavirus infections, such as that by porcine epidemic diarrhea virus (PEDV). In this study, TTSPs including TMPRSS2, HAT, DESC1, and MSPL were tested for their ability to facilitate PEDV replication in Vero cells. Our results showed that TMPRSS2 and MSPL played significant roles in the stages of cell–cell fusion and virus–cell fusion, whereas HAT and DESC1 exhibited weaker effects. This activation may be involved in the interaction between TTSPs and the PEDV S protein, as the S protein extensively co-localized with TMPRSS2 and MSPL and could be cleaved by co-expression with TMPRSS2 or MSPL. Moreover, the use of Vero cells expressing TMPRSS2 and MSPL facilitated PEDV replication in the absence of exogenous trypsin. In sum, we identified two host proteases, TMPRSS2 and MSPL, which may provide insights and a novel method for enhancing viral titers, expanding virus production, and improving the adaptability of PEDV isolates in vitro.
D URING recent years significant advances have been made in using and development of biodegradable polymeric materials for life applications. Degradable polymeric biomaterials are preferred because these materials have specific physical, chemical, biological, biomechanical and degradation properties. Wide ranges of natural or synthetic biopolymers capable of undergoing degradation hydrolytically or enzymatically are being investigated for many applications. This review aimed to provide an overview for the importance of biomaterials, produced or degraded naturally, classification and applications.
The feasibility of Laminaria japonica powder (LJP) combined with cecropin as a dietary supplement to enhance broiler growth performance and immune function was evaluated in this study. In total, 648 oneday-old Arbor Acres broiler chicks were randomly distributed into nine numerically-equal treatment groups: T 1 (control group; fed a basal diet); T 2 (fed the basal diet supplemented with 1% LJP);T 3 (fed the basal diet supplemented with 300mg cecropin/kg); and T 4 ,T 5 ,T 6 ,T 7 ,T 8 and T 9 , individually fed with the dietary supplemented with varying levels of LJP and cecropin). Compared with the control, dietary of LJP or cecropin supplementation slightly improved feed conversion ratio (FCR). However, the dietary supplementation of LJP combined with cecropin significantly improved broiler growth performance during the periods of 1-21,21-42, and 1-42 days (p<0.05).The dietary supplementation of 3% LJP combined with 300 mg/kg cecropin significantly increased FCR, and serum Newcastle disease antibody titers and lymphocyte numbers during the periods of 1-21, 21-42, and 1-42 days (p<0.05). Cecal microorganisms were cultivated and the number of Escherichia coli and Lactobacillus colonies were counted. The dietary supplementation of LJP combined with cecropin remarkably inhibited E. coli growth and increased Lactobacillus growth. The results of this study demonstrate the feasibility of using LJP and cecropin as feed supplement for improving the growth performance and enhancing the immune function of broilers.
We synthesized chitosan grafted with β-cyclodextrin (CD-g-CS) from mono-6-deoxy-6-(p-toluenesulfonyl)-β-cyclodextrin and chitosan. Two different amounts of immobilized β-cyclodextrin (β-CD) on CD-g-CS (QCD: 0.643 × 103 and 0.6 × 102 μmol/g) were investigated. The results showed that the amino contents of CD-g-CS with QCD = 0.643 × 103 and 0.6 × 102 μmol/g were 6.34 ± 0.072 and 9.41 ± 0.055%, respectively. Agar diffusion bioassay revealed that CD-g-CS (QCD = 0.6 × 102 μmol/g) was more active against Staphylococcus xylosus and Escherichia coli than CD-g-CS (QCD = 0.643 × 103 μmol/g). Cell membrane integrity tests and scanning electron microscopy observation revealed that the antimicrobial activity of CD-g-CS was attributed to membrane disruption and cell lysis. Uptake tests showed that CD-g-CS promoted the uptake of doxorubicin hydrochloride by S. xylosus, particularly for CD-g-CS with QCD = 0.6 × 102 μmol/g, and the effect was concentration dependent. CD-g-CS (QCD = 0.6 × 102 and 0.643 × 103 μmol/g) also improved the aqueous solubilities of sulfadiazine, sulfamonomethoxine, and sulfamethoxazole. These findings provide a clear understanding of CD-g-CS and are of great importance for reducing the dosage of antibiotics and antibiotic residues in animal-derived foods. The results also provide a reliable, direct, and scientific theoretical basis for its wide application in the livestock industry.
In this study, the SWKQ series microcomputer automatic incubator was used to study the growth and development of quail in the embryonic stage. Results showed that the embryo shape of became gradually defined as embryo aged. On day 6, the head and body of quail were clearly differentiated, the legs became longer and the wings appeared. At 7 embryo age, the entire embryo of quail was very clear, and the beak has formed. During 3 to 9 days of age, quail embryos length increased quickly, showing a linearly upward trend. At 9 day old, quail embryos length reached 2.2 cm. The regression equation of embryo length to day old was curve regression, giving as following: y=-0.464+0.325x-0.004x 2 , y: embryo length, x: the age of the embryo.
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