Establishment of a rapid real-time fluorescence-based recombinase-aided amplification method for detection of avian infectious bronchitis virus.

Infectious bronchitis (IB) is an acute, highly contagious contact respiratory disease of chickens caused by infectious bronchitis virus (IBV). IBV is very prone to mutation, which brings great difficulties to the prevention and control of the disease. Therefore, there is a pressing need for a method that is fast, sensitive, specific, and convenient for detecting IBV. In this study, a real-time fluorescence-based recombinase-aided amplification (RF-RAA) method was established. Primers and probe were designed based on the conserved regions of the IBV M gene and the reaction concentrations were optimized, then the specificity, sensitivity, and reproducibility of this assay were tested. The results showed that the RF-RAA method could be completed at 39℃ within 20 min, during which the results could be interpreted visually in real-time. The RF-RAA method had good specificity, no cross-reaction with common poultry pathogens, and it detected a minimum concentration of template of 2 copies/µL for IBV. Besides, its reproducibility was stable. A total of 144 clinical samples were tested by RF-RAA and real-time quantitative PCR (qPCR), 132 samples of which were positive and 12 samples were negative, and the coincidence rate of the two methods was 100 %. In conclusion, the developed RF-RAA detection method is rapid, specific, sensitive, reproducible, and convenient, which can be utilized for laboratory detection and clinical diagnosis of IBV.

Development of a colloidal gold immunochromatographic strip for rapid detection of avian coronavirus infectious bronchitis virus.

Avian infectious bronchitis virus (IBV) still causes serious economic losses in the poultry industry. Currently, there are multiple prevalent genotypes and serotypes of IBVs. It is imperative to develop a new diagnosis method that is fast, sensitive, specific, simple, and broad-spectrum. A monoclonal hybridoma cell, N2D5, against the IBV N protein was obtained after fusion of myeloma SP2/0 cells with spleen cells isolated from the immunized Balb/c mice. The N2D5 monoclonal antibody (mAb) and the previously prepared mouse polyclonal antibody against the IBV N protein were used to target IBV as a colloidal gold-mAb conjugate and a captured antibody, respectively, in order to develop an immunochromatographic strip. The optimal pH and minimum antibody concentration in the reaction system for colloidal gold-mAb N2D5 conjugation were pH 6.5 and 30 µg/mL, respectively. Common avian pathogens were tested to evaluate the specificity of the strip and no cross-reaction was observed. The sensitivity of the strip for detecting IBV was 10-1.4522 EID50/mL. The strip showed a broad-spectrum cross-reactive capacity for detecting IBV antigens, including multiple IBV genotypes in China and all of the seven serotypes of IBV that are currently prevalent in southern China. Additionally, the result can be observed within 2 min without any equipment. The throat and cloacal swab samples of chickens that were artificially infected with three IBV strains were tested using the developed strip and the qPCR method; the strip test demonstrated a high consistency in detecting IBV via qPCR gene detection. In conclusion, the immunochromatographic strip that was established is rapid, sensitive, specific, simple, practical, and broad-spectrum; additionally, it has the potential to serve as an on-site rapid detection method of IBV and can facilitate the surveillance and control of the disease, especially in resource-limited areas.

Thermal stability of levopimaric acid and its oxidation products.

Biofuels are renewable alternatives to fossil fuels. Levopimaric acid‒base biofuels have attracted increasing attention. However, their stability remains a critical issue in practice. Thus, there is a strong impetus to evaluate the thermal stability of levopimaric acid. Through thermogravimetry (TG) and a custom-designed mini closed pressure vessel test (MCPVT) operating under isothermal and stepped temperature conditions, we investigated thermal oxidation characteristics of levopimaric acid under oxygen atmosphere. Thin-layer chromatography (TLC) and iodimetry were used to measure the hydrogen peroxides generated by levopimaric acid oxidation. A high pressure differential scanning calorimeter (HPDSC) was used to assess hydroperoxide thermal decomposition characteristics. Gas chromatography-mass spectrometry (GC-MS) was used to characterize the oxidation products. The thermal decomposition kinetics of levopimaric acid were thus elucidated, and a high peroxide value was detected in the levopimaric acid. The decomposition heat (QDSC) and exothermic onset temperature (Tonset) of hydroperoxides were 338.75 J g-1 and 375.37 K, respectively. Finally, levopimaric acid underwent a second-stage oxidation process at its melt point (423.15 K), resulting in complex oxidation products. Thermal oxidation of levopimaric acid could yield potential thermal hazards, indicating that antioxidants must be added during levopimaric acid application to protect against such hazardous effects.

How destination brand experience influences tourist citizenship behavior: Testing mediation of brand relationship quality and moderation effects on commitment.

This study examines the potential predictors of tourist citizenship behavior based on the Stimulus-Organism-Response framework. The studies were conducted in China. Data were collected via questionnaire surveys. Structural equation path modeling and mediation as well as moderation role were used for data analyses. This model was used to test the hypotheses using a sample of 325 individuals with tourism experience in Guangzhou city. The results reveal that tourism destination brand experience and brand relationship quality significantly affect tourist citizenship behavior. Furthermore, the results show that brand relationship quality significantly mediates the relationship between tourism destination brand experience and tourist citizenship behavior and demonstrate that commitment plays a significant moderating role between brand relationship quality and tourist citizenship behavior. This study clearly shows the relationship between tourism destination brand experience, brand relationship quality, and tourist citizenship behavior. Thus, this study contributes to existing tourism studies by identifying gaps and proposing a holistic view to understand tourist citizenship behavior in the tourism industry.

Thermal-birefringence-induced depolarization in a 450 W Ho:YAG MOPA system.

We demonstrated an efficient, high-power Ho:YAG master-oscillator power amplifier (MOPA) system and investigated its thermal-birefringence-induced depolarization. The maximum output power was 450 W with a depolarized power of 32.1 W and depolarization of 0.071 via three power amplifiers. To our knowledge, this is the highest average power generated from a Ho:YAG MOPA system. In theory, a simplified model was built to calculate the depolarization in the amplifier, and the theoretical results agreed with the actual value well. Moreover, the overall optical-to-optical efficiency of the MOPA system was near 60%, and the beam quality M2 factors of s-polarized laser were measured to be ∼ 1.8 at 400 W. In pulse operation, the per pulse energy was ∼ 11 mJ at the pulse repetition frequency of 40 kHz with the corresponding peak power of 220 kW.

113 W Ho:YLF oscillator with good beam quality efficiently pumped by a Tm:YAP laser.

We demonstrate a linearly polarized Tm:YAP slab laser pumped by fiber-coupled laser diodes. The maximum output power is 202 W at 1937.5 nm with a slope efficiency of 47.4% and an optical-to-optical efficiency of 35.6%. The beam quality M2 factors are 10.1 and 8.33 in x and y directions, respectively. Using the Tm:YAP laser as the pump source, the maximum power of the Ho:YLF oscillator is 113 W at 2063.3 nm, corresponding to an optical-to-optical efficiency of 55.9%. In addition, the beam quality factors of the Ho:YLF laser are ∼1.5 at maximum power.

In vivo antiviral effect of plant essential oils against avian infectious bronchitis virus.

BACKGROUND: Infectious bronchitis virus (IBV) leads to huge economic losses in the poultry industry worldwide. The high levels of mutations of IBV render vaccines partially protective. Therefore, it is urgent to explore an effective antiviral drug or agent. The present study aimed to investigate the in vivo anti-IBV activity of a mixture of plant essential oils (PEO) of cinnamaldehyde (CA) and glycerol monolaurate (GML), designated as Jin-Jing-Zi. RESULTS: The antiviral effects were evaluated by clinical signs, viral loads, immune organ indices, antibody levels, and cytokine levels. The infection rates in the PEO-M (middle dose) and PEO-H (high dose) groups were significantly lower than those in the prevention, positive drug, and PEO-L (low dose) groups. The cure rates in the PEO-M and PEO-H groups were significantly higher than those in the prevention, positive drug, and PEO-L groups, and the PEO-M group had the highest cure rate of 92.31%. The symptom scores and IBV mRNA expression levels were significantly reduced in the PEO-M group. PEO significantly improved the immune organ indices and IBV-specific antibody titers of infected chickens. The anti-inflammatory factor levels of IL-4 and IFN-γ in the PEO-M group maintained high concentrations for a long time. The IL-6 levels in the PEO-M group were lower than those in prevention, positive drug, and PEO-L groups. CONCLUSION: The PEO had remarkable inhibition against IBV and the PEO acts by inhibiting virus multiplication and promoting immune function, suggesting that the PEO has great potential as a novel anti-IBV agent for inhibiting IBV infection.

Identification of a novel avian coronavirus infectious bronchitis virus variant with three-nucleotide-deletion in nucleocapsid gene in China.

A novel avian infectious bronchitis virus (IBV) variant, designated as GX-NN160421, was isolated from vaccinated chicken in Guangxi, China, in 2016. Based on analysis of the S1 gene sequence, GX-NN160421 belonged to the New-type 1 (GVI-1) strain. More importantly, three consecutive nucleotides (AAC) deletions were found in the highly conserved structure gene N. The serotype of GX-NN160421 was different from those of the commonly used vaccine strains. The mortality of the GX-NN160421 strain was 3.33%, which contrasted with 50% mortality in the clinical case, but high levels of virus shedding lasted at least 21 days. In conclusion, the first novel IBV variant with three-nucleotide-deletion in the N gene was identified, and this unique variant is low virulent but with a long time of virus shedding, indicating the continuing evolution of IBV and emphasizing the importance of limiting exposure to novel IBV strains as well as extensive monitoring of new IBVs.

Few-moded ultralarge mode area chalcogenide photonic crystal fiber for mid-infrared high power applications.

We demonstrate a novel few-moded ultralarge mode area chalcogenide glass photonic crystal fiber for mid-infrared high power applications. The numerical simulation indicates that the fiber has ultralarge mode areas of ∼10500 µm2 and ∼12000 µm2 for the fundamental mode LP01 and the lowest higher-order mode LP11, respectively. Dual-moded operation is confirmed experimentally at 2 µm, in good agreement with the numerical simulation. By selectively launching technique, low bending loss of 0.7 dB/m, equivalent to 0.55 dB/turn, has been observed in the fiber with a small bending radius of ∼12 cm, indicating excellent bending resistance of the few-moded fiber with such a large mode area. The fiber has been demonstrated to sustain an incident power density up to 150 kW/cm2 under 2-µm CW laser irradiation, showing the potential of the fiber for high-power applications in mid-infrared.

Gain-switched laser diode seeded TDFA with 409 W picosecond pulses and 142 W spectrally flat supercontinuum output.

We report on a high-power picosecond all-fiber Tm-doped fiber amplifier (TDFA) seeded by a gain-switched laser diode (LD) in the 2 µm spectral range. A total average output power of 409 W (304 ps) has been generated at 320 MHz of repetition rate with 10 dB bandwidth of ~48 nm centered at 1970 nm. Over 140 W of spectrally flat supercontinuum (SC) output has been produced at 40 MHz of repetition rate with optimized fiber length. The 10 dB spectral bandwidth was 615 nm, ranging from 1965 to 2580 nm. The prospects for further power scaling of LD seeded ~2 µm picosecond all-fiber sources are discussed.

Ho3+:Y2O3 ceramic laser generated over 113 W of output power at 2117 nm.

We demonstrate efficient laser operation using a Ho:Y2O3 ceramic with low scattering loss and excellent optical quality fabricated in-house through an improved technique in support of high-power laser operation. The Ho:Y2O3 ceramic laser was in-band pumped by a Tm-fiber laser of 1931 nm at room temperature. With an optically polished but uncoated sample of 0.5 at. % Ho3+ doping, up to 113.6 W continuous-wave output power at 2117 nm has been generated with a slope efficiency of 55.6% with respect to the absorbed pump power. To the best of our knowledge, this is the highest output power achieved with a Ho-doped ceramic.