Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Detection of Raccoon Dog in Meat Mixtures

Raccoon dog (Nyctereutes procyonoides) is an economically important animal used for fur production, but consuming its meat is injurious to human health. Currently, no rapid and sensitive method for detecting raccoon dog meat in meat mixtures is available. In this study, we developed an easily applicable, rapid, and economically feasible method for identifying the presence of raccoon dog in meat mixtures based on loop-mediated isothermal amplification (LAMP). Four sets of LAMP primers were tested at different temperatures, and the primers that worked best at 62°C (set 2) were determined. In the LAMP assay, there was no cross-reactivity with the meat procured from other species of animals and the detection limit of DNA concentration was 0.1 pg·μL−1, slightly higher than TaqMan real-time PCR (0.01 pg·μL−1), but sensitivity of 0.1 pg·μL−1 complies with most requirements of routine analysis. Moreover, by the LAMP method, the meat mixtures containing more than 0.5% of the raccoon dog component were directly detected (without DNA extraction) in the supernatant isolated from the meat mixtures after performing repeated cycles of thawing and freezing of minced meat mixtures. Our results show that LAMP assay is a valuable, straightforward, and sensitive detection tool for identification of raccoon dog meat in mixtures

A Correlation Study between Two Adjacent Same-Meridian Acupoints after Laser-Needle Acupuncture with Optical Coherence Tomography and Diffuse Reflectance Spectra

This study is to investigate the correlations among Sanjian (LI3), Hegu (LI4), and Yangxi (LI5) acupoints and their corresponding nonacupoints on the Yangming Large Intestine Meridian of Hand before and after laser irradiation using optical coherence tomography (OCT) and diffuse reflectance spectra. The experiment was conducted on 10 healthy people. A 658 nm laser with 50 mW output power was used for irradiating LI4, LI5 acupoints and their corresponding nonacupoints. As to LI4 acupoint with laser irradiation for duration of 15 or 45 minutes, the OCT backscattered light intensities of LI4 and LI5 acupoints increased significantly, and the reflectance intensities (RIs) of the LI3, LI4, and LI5 acupoints decreased significantly. As to LI5 acupoint with laser irradiation for duration of 15 or 45 minutes, the changes of OCT backscattered light intensities of the corresponding irradiated acupoint and LI4 acupoint increased significantly, and the RIs decreased significantly. However, the OCT backscattered light intensities and RIs for their nonacupoints were almost not changed. The results show that an association exists between two adjacent same-meridian acupoints on the same meridian after laser-needle acupuncture to some extent

Maternal consumption of fish oil protected breast-fed piglets against Escherichia coli lipopolysaccharide-induced damage through reshaping of intestinal fatty acids profile

It has been well documented that n-3 polyunsaturated fatty acids (n-3 PUFA) can alleviate inflammation caused by Escherichia coli (E. coli) lipopolysaccharides (LPS), the etiologic agents that causing yellow or white dysentery in young pigs. However, it remains unclear whether the increase in n-3 PUFA availability could enhance the ability of nursery pigs to resist invasion by E. coli. LPS. Twenty-four 21-day-old female piglets, each two of them from the same sow fed the beef tallow (BT) or fish oil (FO) diets, were allocated into four treatment groups: BT-CON, piglets from the BT-fed sows and intraperitoneally injected with saline (9 g/L); BT-LPS, piglets from the BT-fed sows and injected with LPS (100 μg/kg body weight); FO-CON, piglets from the FO-fed sows and injected with saline; FO-LPS, piglets from the FO-fed sows and injected with LPS. Following 2 h of LPS challenge, the magnitudes of increase in body temperature approached to a marked (p < 0.01) difference between the BT-CON and BT-LPS piglets, whereas the dramatic (p < 0.01) difference between the FO-CON and FO-LPS piglets was only observed at 4 h post LPS challenge. The body temperature averaged across the time points evaluated was about 0.2°C lower (p < 0.05) in the FO group than in the BT group. The FO group had lower (p < 0.05) mean corpuscular hemoglobin concentration, lower increase in serum interleukin (IL)-1β (p < 0.10) and IL-8 (p < 0.05) levels, higher (p < 0.01) serum albumin concentration, and higher (p = 0.10) ratios of jejunum villus height to crypt depth than the BT group. The FO group had much higher (p < 0.0001) ileal content of C20:5n3, C24:0, and C22:6n3, which were 2–4 times the content of the BT group. LPS challenge resulted in decreased (p < 0.05) intestinal C20:1 and C20:5n3 content, and the decrease (p < 0.05) in intestinal C20:3n6 and C24:1 content was observed in the BT-LPS piglets rather than in the FO-LPS piglets. Taken together, this study indicated that maternal consumption of fish oil protected breast-fed piglets against E. coli LPS-induced damage through reshaping of intestinal fatty acids profile, which sheds new light on the development of nutritional strategies to enhance the ability of young pigs to resist E. coli invasion

Antimicrobial susceptibility and minimum inhibitory concentration distribution of common clinically relevant non-tuberculous mycobacterial isolates from the respiratory tract

OBJECTIVE: To determine the minimum inhibitory concentration (MIC) distribution of antibacterial drugs and the susceptibility of non-tuberculous mycobacterial (NTM) isolates to provide a reference basis for the clinical selection of an effective starting regimen. METHODS: The common clinical isolates of NTM in the respiratory tract, which met the standards of the American Thoracic Society for NTM lung disease, were collected. The MICs of 81 isolates were determined using the microbroth dilution method (Thermo Fisher Scientific, USA), as recommended by the Clinical and Laboratory Standards Institute, USA. RESULTS: Included were 43 Mycobacterium avium complex (MAC) strains, 24 M. abscessus complex (MAB) strains, and 14 M. kansasii strains. The sensitivity rates of MAC to clarithromycin and amikacin were 81.4% and 79.1%, respectively, while the sensitivity rates to linezolid and moxifloxacin were only 20.9% and 9.3%; the MIC of rifabutin was the lowest (MIC50% was just 2 mu g/mL). After incubation for 3-5 days, the sensitivity rate of MAB to clarithromycin was 87.5%; this decreased to 50% after 14 days' incubation. Most of them were susceptible to amikacin (91.6%), and most were resistant to moxifloxacin (95.8%), ciprofloxacin (95.8%), imipenem (95.8%), amoxicillin/clavulanate (95.8%), tobramycin (79.1%), doxycycline (95.8%) and trimethoprim/sulfamethoxazole (95.8%). intermediate (83.3%) and resistant (16.7%) to cefoxitin. The susceptibility to linezolid was only 33.3%. The sensitivity and resistance breakpoints of tigecycline were set to <= 0.5 and >= 8 mu g/mL, respectively, and the sensitivity and resistance rates were 50% and 0%, respectively. M. kansasii was susceptible to clarithromycin, amikacin, linezolid, moxifloxacin, rifampicin and rifabutin (100%). DISCUSSION: In Wenzhou, clarithromycin, amikacin and rifabutin have good antibacterial activity against MAC, while linezolid and moxifloxacin have high resistance. Amikacin and tigecycline have strong antibacterial activity against MAB, while most other antibacterial drugs are resistant to varying degrees. Most antibacterial drugs are susceptible to M. kansasii and have good antibacterial activity. Conclusion: The identification of NTM species and the detection of their MICs have certain guiding values for the treatment of NTM lung disease. KEY MESSAGE: The three most common respiratory non-tuberculous mycobacterial (NTM) isolates with clinical significance in the Wenzhou area were tested for drug susceptibility. The broth microdilution method was used to determine the minimum inhibitory concentration distribution of antibacterial drugs and the susceptibility of NTM isolates to provide a reference basis for the clinical selection of an effective starting regimen

Drug resistance in drug-resistant tuberculosis patients with and without diabetes mellitus: a comparative analysis

Abstract Background This dual burden of tuberculosis (TB) and diabetes mellitus (DM) has become a global public health concern. This study aims to compare drug resistance in drug-resistant tuberculosis (DR-TB) patients with and without DM and analyse the risk factors of multidrug-resistant tuberculosis (MDR-TB). Methods A total of 893 DR-TB patients were admitted to Wenzhou Central Hospital between January 2018 and December 2022. After excluding 178 cases with incomplete clinical and laboratory data, 715 patients were included in the study. These patients were then categorized into two groups based on the presence of type 2 DM: the DM group (160 cases) and the non-DM group (555 cases). Demographic information, baseline clinical characteristics, laboratory and imaging test results, clinical diagnoses, and other relevant data were collected for analysis. Statistical analysis was conducted on demographic information, clinical parameters, drug resistance spectrum, and risk factors for multidrug resistance. Results In both the DM and non-DM groups, the order of resistance to first-line anti-tuberculosis drugs is isoniazid, streptomycin, rifampicin, and ethambutol. There is no significant difference in the proportion of mono-resistant tuberculosis, polydrug-resistant tuberculosis, and multidrug-resistant tuberculosis between the two groups (P > 0.05). The prevalence of MDR-TB in both groups shows a downward trend between 2018 and 2022, but the trend is not statistically significant (P > 0.05). Among patients without DM, residence in rural areas, retreatment of tuberculosis, pulmonary cavity, and uric acid ≥ 346 µmol/L are identified as independent risk factors for MDR-TB. Among patients with DM, residence in rural areas, retreatment of tuberculosis, pulmonary cavity, and HbA1c ≥ 9.8% were identified as independent risk factors for MDR-TB. Conclusion Isoniazid is the most resistant drug among DR-TB patients with and without DM. There is no statistically significant difference in drug resistance patterns between the two groups. Some progress has been made in the prevention and control of DR-TB in this area, but the effect is not very significant. There are differences in the risk factors of MDR-TB between patients with and without DM

Evaluation of WHO catalog of mutations and five WGS analysis tools for drug resistance prediction of Mycobacterium tuberculosis isolates from China

ABSTRACT The continuous advancement of molecular diagnostic techniques, particularly whole-genome sequencing (WGS), has greatly facilitated the early diagnosis of drug-resistant tuberculosis patients. Nonetheless, the interpretation of results from various types of mutations in drug-resistant-associated genes has become the primary challenge in the field of molecular drug-resistance diagnostics. In this study, our primary objective is to evaluate the diagnosis accuracy of the World Health Organization (WHO) catalog of mutations and five WGS analysis tools (PhyResSE, Mykrobe, TB Profiler, Gen-TB, and SAM-TB) in drug resistance to 10 anti-Mycobacterium tuberculosis (MTB) drugs. We utilized the data of WGS collected between 2014 and 2017 in Zhejiang Province, consisting of 110 MTB isolates as detailed in our previous study. Based on phenotypic drug susceptibility testing (DST) results using the proportion method on Löwenstein-Jensen medium with antibiotics, we evaluated the predictive accuracy of genotypic DST obtained by these tools. The results revealed that the WHO catalog of mutations and five WGS analysis tools exhibit robust predictive capabilities concerning resistance to isoniazid, rifampicin, ethambutol, streptomycin, amikacin, kanamycin, and capreomycin. Notably, Mykrobe, SAM-TB, and TB Profiler demonstrate the most accurate predictions for resistance to pyrazinamide, prothionamide, and para-aminosalicylic acid, respectively. These findings are poised to significantly guide and influence future clinical treatment strategies and resistance monitoring protocols.IMPORTANCEWhole-genome sequencing (WGS) has the potential for the early diagnosis of drug-resistant tuberculosis. However, the interpretation of mutations of drug-resistant-associated genes represents a significant challenge as the amount and complexity of WGS data. We evaluated the accuracy of the World Health Organization catalog of mutations and five WGS analysis tools in predicting drug resistance to first-line and second-line anti-TB drugs. Our results offer clinicians guidance on selecting appropriate WGS analysis tools for predicting resistance to specific anti-TB drugs

Microstructure and Property Evolutions of Q345B Steel during Ultrasonic Shot Peening

In this paper, the surface of Q345B steel was strengthened using ultrasonic shot peening (USP) technology. Through the adjustment of USP time, power and distance, the surface morphology, roughness and microhardness of the USPed samples were measured using scanning electron microscopy (SEM), energy dispersive analysis (EDS), a roughmeter and a microhardness tester. At the same time, the corrosion behavior of USPed samples was observed and analyzed using software simulation calculation and an immersion experiment on the dynamic polarization curve. Through tests and characterization, the influence of different USP process parameters on Q345B steel and the relationship between them were explored. The original intention of this research was to obtain better parameters to improve both the strength and corrosion resistance of the material. The results indicated that, with the increase in the USP time and power and the decrease in the USP distance, the surface roughness, the thickness of the deformed layer and the microhardness of the samples increased at first and then stabilized, and an obvious delamination phenomenon and chemical composition difference appeared between the deformed layer and the substrate. It was shown that a longer USP time and a shorter USP distance caused spalling and cracks on a substrate surface, resulting in the corrosion becoming more serious. However, with the increase in the USP power, the corrosion resistance of the sample improved

Preparation of Nanocellulose-Based Aerogel and Its Research Progress in Wastewater Treatment

Nowadays, the fast expansion of the economy and industry results in a considerable volume of wastewater being released, severely affecting water quality and the environment. It has a significant influence on the biological environment, both terrestrial and aquatic plant and animal life, and human health. Therefore, wastewater treatment is a global issue of great concern. Nanocellulose’s hydrophilicity, easy surface modification, rich functional groups, and biocompatibility make it a candidate material for the preparation of aerogels. The third generation of aerogel is a nanocellulose-based aerogel. It has unique advantages such as a high specific surface area, a three-dimensional structure, is biodegradable, has a low density, has high porosity, and is renewable. It has the opportunity to replace traditional adsorbents (activated carbon, activated zeolite, etc.). This paper reviews the fabrication of nanocellulose-based aerogels. The preparation process is divided into four main steps: the preparation of nanocellulose, gelation of nanocellulose, solvent replacement of nanocellulose wet gel, and drying of nanocellulose wet aerogel. Furthermore, the research progress of the application of nanocellulose-based aerogels in the adsorption of dyes, heavy metal ions, antibiotics, organic solvents, and oil-water separation is reviewed. Finally, the development prospects and future challenges of nanocellulose-based aerogels are discussed

Nanocellulose and Cellulose Making with Bio-Enzymes from Different Particle Sizes of Neosinocalamus Affinis

Cellulose is one of the most abundant, widely distributed and abundant polysaccharides on earth, and is the most valuable natural renewable resource for human beings. In this study, three different particle sizes (250, 178, and 150 μm) of Neosinocalamus affinis cellulose were extracted from Neosinocalamus affinis powder using bio-enzyme digestion and prepared into nanocellulose (CNMs). The cellulose contents of 250, 178, and 150 μm particle sizes were 53.44%, 63.38%, and 74.08%, respectively; the crystallinity was 54.21%, 56.03% and 63.58%, respectively. The thermal stability of cellulose increased gradually with smaller particle sizes. The yields of CNMs for 250, 178, and 150 μm particle sizes were 14.27%, 15.44%, and 16.38%, respectively. The results showed that the Neosinocalamus affinis powder was successfully removed from lignin, hemicellulose, and impurities (pectin, resin, etc.) by the treatment of bio-enzyme A (ligninase:hemicellulose:pectinase = 1:1:1) combined with NH3·H2O and H2O2/CH3COOH. Extraction of cellulose from Neosinocalamus affinis using bio-enzyme A, the smaller the particle size of Neosinocalamus affinis powder, the more cellulose content extracted, the higher the crystallinity, the better the thermal stability, and the higher the purity. Subsequently, nanocellulose (CNMs) were prepared by using bio-enzyme B (cellulase:pectinase = 1:1). The CNMs prepared by bio-enzyme B showed a network structure and fibrous bundle shape. Therefore, the ones prepared in this study belong to cellulose nanofibrils (CNFs). This study provides a reference in the extraction of cellulose from bamboo using bio-enzymes and the preparation of nanocellulose. To a certain extent, the utilization of bamboo as a biomass material was improved

Extraction of Corn Bract Cellulose by the Ammonia-Coordinated Bio-Enzymatic Method

This study explored a green and efficient method for cellulose extraction from corn bract. The cellulose extraction by the CHB (CH3COOH/H2O2/Bio-enzyme) method and the N-CHB (NH3·H2O-CH3COOH/H2O2/Bio-enzyme) method were compared and analyzed. The effect of ammonia pretreatment on cellulose extraction by bio-enzymatic methods was discussed. The results showed that ammonia promoted the subsequent bio-enzymatic reaction and had a positive effect on the extraction of cellulose. Sample microstructure images (SEM) showed that the cellulose extracted by this method was in the form of fibrous bundles with smooth surfaces. The effect of different pretreatment times of ammonia on cellulose was further explored, and cellulose was characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and thermogravimetric (TG) analysis. The results showed that the N3h-CHB (NH3·H2O 50 °C 3 h, CH3COOH/H2O2 70 °C 11 h, Bio-enzyme 50 °C 4 h) method was the best way to extract cellulose in this study. FTIR showed that most of the lignin and hemicellulose were removed. XRD showed that all the cellulose extracted in this study was type I cellulose. TG analysis showed that the cellulose was significantly more thermally stable, with a maximum degradation temperature of 338.9 °C, close to that of microcrystalline cellulose (MCC). This study provides a reference for the utilization of corn bract and offers a new technical route for cellulose extraction