Fortification of yogurt with mulberry leaf extract: Effects on physicochemical, antioxidant, microbiological and sensory properties during 21-days of storage.

Mulberry leaves are medicinal and edible, with many physiological functions. To improve the potential function of yogurt, the effects of mulberry leaf extract (MLE) incorporation on the fermentation kinetics, physicochemical, antioxidant properties, and sensory parameters of yogurt were evaluated. The results showed that 0.1-0.3 % MLE improved the acidification rate and shortened the fermentation process. The addition of MLE significantly increased the values of total titratable acids, water holding capacity (WHC), total phenolic content and antioxidant capacities of the yogurt (p＜0.05). Specifically, the WHC values of 0.1 % MLE added yogurt were 1.33-1.41 times that of the control over 21 days of storage. In addition, MLE changed the texture and sensory quality of yogurt, resulting in light green, more stable products. Compared to the control, the yogurt with an appropriate concentration of MLE (0.1 % and 0.2 %) showed stable microbiological properties, and the survival of lactic acid bacteria in the yogurt was able to maintain a stable probiotic count of 108 CFU/g over 21 days of shelf life. The yogurt containing 0.1 % MLE achieved a good balance between the physicochemical and sensory qualities of the yogurt, and the use of MLE as an ingredient in yogurt production was a step towards the development of healthier dairy products.

Development and Application of Colloidal Gold Test Strips for the Rapid Detection of Canine Brucellosis.

Brucellosis is a global problem, with the causative agent being the genus Brucella. B. canis can cause undulant fever in dogs, which is a zoonotic disease that can spread not only among dogs but also to humans. This poses a public health threat to society. In this study, a rapid and straightforward immune colloidal gold test strip was developed for the diagnosis of canine brucellosis through the detection of anti-LPS antibodies in serum samples. Rabbit anti-canine IgG conjugated with colloidal gold was employed as the colloidal gold-labeled antibody. The extracted high-purity R-LPS was employed as the capture antigen in the test line (T-line), while goat anti-rabbit IgG was utilized as the capture antibody in the control line (C-line). The colloidal gold strip exhibited high specificity in the detection of brucellosis, with no cross-reaction observed with the common clinical canine diseases caused by Canine coronavirus (CCV), Canine distemper virus (CDV), and Canine parvovirus (CPV). In comparison to the commercial iELISA kit, the sensitivity and specificity of the colloidal gold test strip were found to be 95.23% and 98.76%, respectively. The diagnostic coincidence rate was 98.47%. The findings of this study indicate that colloidal gold test strips may be employed as a straightforward, expeditious, sensitive, and specific diagnostic instrument for the identification of canine brucellosis, particularly in resource-limited regions.

Establishment and application of quadruple fluorescence quantitative RT-PCR method for the identification of waterfowl astrovirus.

Avian astrovirus can infect a variety of poultry species and cause viral diarrhea, with a wide epidemic range strong pathogenicity and a high incidence. Among them, Duck astrovirus 3(DAstV-3), Duck astrovirus 4(DAstV-4), Goose astrovirus 1(GoAstV-1) and Goose astrovirus 2(GoAstV-2) are four types of astroviruses newly discovered in waterfowl in recent years. In order to realize the rapid detection of these four kinds of waterfowl stellate viruses, specific primers and probes were engineered to target a highly conserved region of ORF1b gene of DAstV-3, GoAstV-1 and GoAstV-2 and the ORF2 gene of DAstV-4, and a quadruple fluorescence quantitative RT-PCR method was developed. The results indicate that the method established in this study has good specificity and no cross reactivity with other pathogens. This method can detect viruses with a minimum concentration of 1 × 101 copies/µL for DAstV-4, GoAstV-1 and GoAstV-2, respectively, while the minimum concentration for DAstV-3 is 1 × 102 copies/µL. Compared with the routinely used RT-PCR method, the limit of detection by the multiplex RT-PCR lower. Both intra- and inter-assay variability tests revealed excellent reproducibility. This method was then used to analyze 269 field samples, and the results were verified by genome sequencing. In conclusion, this study presents a sensitive, accurate, and specific method for detecting DAstV-3, DAstV-4, GoAstV-1, and GoAstV-2 in a single reaction, enabling the monitoring and differential diagnosis of these four types of waterfowl astroviruses.

Evaluation of the mechanistic basis for the antibacterial activity of ursolic acid against Staphylococcus aureus.

The antibiotics are generally regarded as the first choice approach to treat dairy mastitis, targeting the public health problems associated with the food safety and the emergence of antibioticresistant bacteria. The objective of the study was to evaluate the antibacterial efficacy of ursolic acid (UA) when used to treat Staphylococcus aureus and other isolates associated with bovine mastitis and to clarify the mechanistic basis for these effects. The bacteriostatic properties of UA extracted from Rosmarinus officinalis L. at four different purity levels were assessed by calculating minimum inhibitory concentration (MIC) values, while the synergistic effects of combining 98% UA with antibiotics were evaluated by measuring the fractional inhibitory concentration index (FICI). Changes in biofilm formation and the growth curves of the clinical isolates were assessed to clarify the bacteriostatic effect of UA. Furthermore, the cell wall integrity, protein synthesis, and reactive oxygen species (ROS) production were assessed to determine the antibacterial mechanism of UA treatment. Ultimately, UA was revealed to exhibit robust activity against Gram-positive bacteria including S. aureus (ATCC 25923), Streptococcus dysgalactiae (ATCC27957), Streptococcus agalactiae (ATCC13813), Enterococcus faecalis (ATCC29212), and Streptococcus mutans (ATCC25175). However, it did not affect Escherichia coli (ATCC 25922). The MIC values of UA preparations that were 98, 50, 30, and 10% pure against S. aureus were 39, 312, 625, and 625 µg/mL, respectively, whereas the corresponding MIC for E. coli was >5,000 µg/mL. The minimum bactericidal concentrations of 98% UA when used to treat three clinical S. aureus isolates (S4, S5, and S6) were 78, 78, and 156 µg/mL, respectively. Levels of biofilm formation for clinical S. aureus isolates decreased with increasing 98% UA concentrations. Above the MIC dose, UA treatment resulted in the dissolution of bacterial cell walls and membranes, with cells becoming irregularly shaped and exhibiting markedly impaired intracellular protein synthesis. S. aureus treated with 98% UA was able to rapidly promote intracellular ROS biogenesis. Together, these data highlight the promising utility of UA as a compound that can be used together with other antibiotics for the treatment of infections caused by S. aureus.

Production of γ-aminobutyric acid-enriched sourdough bread using an isolated Pediococcus pentosaceus strain JC30.

A γ-aminobutyric acid (GABA)-producing strain JC30 was isolated from traditional kimchi, which was identified as Pediococcus pentosaceus by 16S rDNA sequencing. P. pentosaceus JC30 was highly tolerant to acid, bile salt, and high temperatures. The survival rate of JC30 in MRS medium (pH 2.5) for 3 h was 60.96 %. Furthermore, the survival rate of JC30 in MRS medium with 3 mg/mL bile salt for 24 h was 86.62 %. The survival rate of JC30 in MRS medium at 56 °C and 58 °C for 10 min was 97.17 % and 78.20 %, respectively. When 2 % v/v JC30 (8.0 log10 CFU/mL) was added to prepare sourdough and the sourdough was then used to make bread, the bread had a higher specific volume (5.13 ± 0.12 mL/g) and GABA content (3.32 ± 0.04 mg/g DW) than the control.

Photoselectively modulating main products by changing the wavelength of visible light over D-π-A-D conjugated polymers.

The diversity of catalytic products determines the difficulty of selective product modulation, which usually relies on adjusting the catalyst and reaction conditions to obtain different main products selectively. Herein, we synthesized D-π-A-D conjugated organic polymers (TH-COP) using cyclotriphosphonitrile, alkyne, 2H-benzimidazole, and sulfur units as electron donors, π bridges, electron acceptors, and electron donors, respectively. TH-COP exhibited excellent photoinduced carrier separation and redox ability under different visible light wavelengths, and the main products of its CO2 reduction are CH4 (1000.0 µmol g-1) and CO (837.0 µmol g-1) under 400-420 nm and 420-560 nm, respectively. In addition, TH-COP could completely convert phenylmethyl sulfide to methyl phenyl sulfone at 400-420 nm and diphenyl disulfide at 480-485 nm in yields up to 95 %. This study presents a novel strategy for the targeted fabrication of various main products using conjugated polymers by simply changing the wavelength range of visible light.

Rapid detection of H5 subtype avian influenza virus using CRISPR Cas13a based-lateral flow dipstick.

Due to its high mortality rate, highly pathogenic avian influenza (HPAI), a notifiable animal illness designated by the World Organisation for Animal Health (WOAH), has caused enormous financial losses to the poultry sector. The H5 subtype of avian influenza virus (H5-AIV) is regarded as the most common highly pathogenic avian influenza virus (HPAIV) that threatens public health and safety. Virus isolation and reverse transcription quantitative PCR (RT-qPCR) are usually used to detect H5-AIV and are important for the timely diagnosis and control of H5-AIV. However, these methods are time-consuming and require a significant amount of effort. In this study, we established a recombinase-aided amplification (RAA) combined with CRISPR-Cas13a and lateral flow dipstick (LFD) assay for the detection of H5-AIV. The results showed that the process can be completed within 40 min at 37°C. The method had a detection limit of 0.1 copy/µL, which was comparable to the RT-qPCR. There was no cross-reactivity with H3-AIV, H7-AIV, H9-AIV, H10-AIV, IBV, NDV, RVA and DAstV. The kappa value of RT-RAA-Cas13a-LFD and RT-qPCR in 380 clinical samples was 0.89 (κ>0.75). In conclusion, we established a convenient, efficient and accurate method to detect H5-AIV, and the results can be visualized and interpreted using LFD, which can be adapted to the needs of grassroots laboratories and field-deployable assays. This approach provides a new perspective for clinical H5-AIV diagnosis and has great potential for application in clinical quarantine of the poultry farming.

Ursolic acid: biological functions and application in animal husbandry.

Ursolic acid (UA) is a plant-derived pentacyclic triterpenoid with 30 carbon atoms. UA has anti-inflammatory, antioxidative, antimicrobial, hepato-protective, anticancer, and other biological activities. Most studies on the biological functions of UA have been performed in mammalian cell (in vitro) and rodent (in vivo) models. UA is used in animal husbandry as an anti-inflammatory and antiviral agent, as well as for enhancing the integrity of the intestinal barrier. Although UA has been shown to have significant in vitro bacteriostatic effects, it is rarely used in animal nutrition. The use of UA as a substitute for oral antibiotics or as a novel feed additive in animal husbandry should be considered. This review summarizes the available data on the biological functions of UA and its applications in animal husbandry.

Characteristics of Mulberry Leaf Powder Enriched With γ-Aminobutyric Acid and Its Antioxidant Capacity as a Potential Functional Food Ingredient.

To improve the functional properties of mulberry leaves, γ-aminobutyric acid (GABA) enrichment treatments were applied. The results showed that the combined treatment of sodium glutamate immersion, cold shock, and anoxic significantly increased the GABA content. HPLC analysis displayed that the quantity of some active phenolics was significantly increased after the treatment. The GABA-enriched mulberry leaf powders were subsequently prepared, and it was found that as the particle size decreased, their water and oil holding capacity and their swelling power decreased, while the angle of repose increased. The dissolution rate of GABA and total phenolics increased as the particle size decreased. Optical observations and SEM results revealed that the fiber structures of the particles were gradually destroyed as the particle size decreased. Further, FTIR analysis showed that the active compounds in the powders were not destroyed. M400 and M140 powder showed the maximum DPPH radical scavenging ability and AGEs inhibition capacity, respectively. Additionally, adding the powders effectively alleviated the staling of bread without any significant effect on taste.

Exogenous GABA improves the antioxidant and anti-aging ability of silkworm (Bombyx mori).

Silkworm (Bombyx mori) was used to explore the anti-aging ability of Gamma-aminobutyric acid (GABA) and its potential mechanism. 0.086∼8.6 mM GABA solutions were sprayed on mulberry leaves to raise silkworms. The results showed that GABA increased the fecundity of adult silkworms. And in the larva silkworms on the 3th day of the 5th instar, GABA significantly reduced the trehalose content in the hemolymph, the triglycerides and glycogen levels in the fat body, while sharply increased the NAD+/NADH level in the fat body. GABA significantly increased the GSH content and activities of SOD and CAT, while reduced the level of MDA. Furthermore, GABA reduced the mRNA expression of BmRpd3, Bmchico and BmAkh2, while increased the mRNA expression of BmAMPK. In summary, GABA has anti-aging potential by playing roles in energy homeostasis, reducing carbohydrate and lipid level, increasing anti-oxidative capacity, and regulating mRNA expression of longevity-related genes.

Virulence factors and molecular characteristics of Shigella flexneri isolated from calves with diarrhea.

BACKGROUND: The natural hosts of Shigella are typically humans and other primates, but it has been shown that the host range of Shigella has expanded to many animals. Although Shigella is becoming a major threat to animals, there is limited information on the genetic background of local strains. The purpose of this study was to assess the presence of virulence factors and the molecular characteristics of S. flexneri isolated from calves with diarrhea. RESULTS: Fifty-four S. flexneri isolates from Gansun, Shanxi, Qinghai, Xinjiang and Tibet obtained during 2014 to 2016 possessed four typical biochemical characteristics of Shigella. The prevalences of ipaH, virA, ipaBCD, ial, sen, set1A, set1B and stx were 100 %, 100 %, 77.78 %, 79.63 %, 48.15 %, 48.15 and 0 %, respectively. Multilocus variable number tandem repeat analysis (MLVA) based on 8 variable number of tandem repeat (VNTR) loci discriminated the isolates into 39 different MLVA types (MTs), pulsed field gel electrophoresis (PFGE) based on NotI digestion divided the 54 isolates into 31 PFGE types (PTs), and multilocus sequence typing (MLST) based on 15 housekeeping genes differentiated the isolates into 7 MLST sequence types (STs). CONCLUSIONS: The findings from this study enrich our knowledge of the molecular characteristics of S. flexneri collected from calves with diarrhea, which will be important for addressing clinical and epidemiological issues regarding shigellosis.

Exploring the Prevalence and Distribution Patterns of Antibiotic Resistance Genes in Bovine Gut Microbiota Using a Metagenomic Approach.

Antibiotic resistance genes (ARGs) have become recognized contaminants and pose a high public health risk. The animal gut microbiota is a reservoir of ARGs, but the knowledge of the origin and dissemination of ARGs remains unclear. In this study, we provide a comprehensive profile of ARGs and mobile genetic elements in the gut microbiota from 30 bovines to study the impact of modern antibiotics on resistance. A total of 42 ARG types were detected by annotating the metagenomic sequencing data from Comprehensive Antibiotic Resistance Database (CARD). We found that the diversity and abundance of ARGs in individual yaks were significantly lower than those in dairy and beef cattle (p < 0.0001). The results of heat map and single nucleotide polymorphism clustering suggest that ARGs from dairy and beef cattle are more similar, whereas those from yaks cluster separately. The long-term use of antibiotics may contribute to this difference, suggesting that antibiotic consumption is the main cause of ARG prevalence. Furthermore, abundant insertions were also found in this study, signifying a strong potential for horizontal transfer of ARGs among microbes, especially pathogens.

Epidemic and molecular characterization of fluoroquinolone-resistant Shigella dysenteriae 1 isolates from calves with diarrhea.

BACKGROUND: The widespread distribution of antimicrobial-resistant Shigella has become a recurrent challenge in many parts of the developing world. Previous studies indicate that the host of Shigella has expanded from humans to animals. This study aimed to investigate the prevalence of fluoroquinolone resistance and associated molecular characterization of S. dysenteriae 1 isolated from calves. RESULTS: All 38 unduplicated S. dysenteriae 1 isolates were collected from calves in Gansu Province from October 2014 to December 2016. According to MLST and PFGE analysis, these isolates were separated into 4 and 28 genotypes, respectively. The most common STs identified were ST228 (34.21%, 13/38) and ST229 (39.47%, 15/38), which were first found in the present study. All isolates harbored virulence genes, and the incidence of the seven virulence genes were ipaH (100%), ipaBCD (92.11%), stx (73.68%), ial (57.89%), sen (28.95%), set1A and set1B (0%). According to the results of antimicrobial susceptibilities, 76.32% (29/38) were resistant to fluoroquinolone and showed multidrug resistance. In a study on the polymorphism of quinolone resistance-determining region (QRDR) of gyrA/B and parC/E genes, we identified two mutations in gyrA (Ser83 → Leu and Asp87 → Asn) and parC (Ser80 → Ile and Ser83 → Leu), respectively. Among them, 55.17% (16/29) of resistant strains had the gyrA point mutations (Ser83 → Leu) and parC point mutation (Ser83 → Leu). Moreover, 41.38% (12/29) of isolates had all five point mutations of gyrA and parC. In addition, the prevalence of the plasmid-mediated quinolone resistance (PMQR) determinant genes was also investigated. All 29 fluoroquinolone-resistant isolates were positive for the aac (6')-Ib-cr gene but negative for qepA, except for SD001. In addition, only 6 (20.69%, 6/29) isolates harbored the qnr gene, including two with qnrB (6.90%, 2/29) and four with qnrS (13.79%, 4/29). CONCLUSION: Given the increased common emergence of multidrug resistant isolates, uninterrupted surveillance will be necessary to understand the actual epidemic burden and control this infection.

Restraint stress induces uterine microenvironment disorder in mice during early pregnancy through the ß2-AR/cAMP/PKA pathway.

During pregnancy, uterus undergoes the environment adaptation as part of a program of development. In the world, one in four people worldwide suffer from mental illness, especially pregnant women. ß-Adrenergic receptor (ß-AR) is an important regulator that converts environmental stimuli into intracellular signals in mice uterus. CD-1 (ICR) mice undergone restraint stress, which was a case in model to simulate the psychological stress. The plasma and implantation sites in uterus were obtained and examined. PCR analysis demonstrated that ß2-AR expression levels in embryo day (E) 3, 5 and 7 were kept at a significantly higher level (p < 0.05) under restraint stress and higher than ß1-AR and ß3-AR in different gestation ages. The ß2-AR protein levels were obviously increased (p < 0.05) due to the markedly elevated norepinephrine (NE) concentration (p < 0.05). In our previous study, restraint stress can induce the apoptosis and inflammation. Also, the matrix metalloprotein-9 (MMP-9) was decreased significantly (p < 0.05) under restraint stress. Meanwhile, Caspase3, p-NF-κB p65 and p-ERK1/2 were obviously increased (p < 0.05) in the work. In vitro studies showed that the p-ERK1/2 and Caspase-3 levels were raised (p < 0.05) after ß2-AR was activated. However, they were decreased when PKA was blocked. The protein levels of Caspase-3 were reduced when ERK and NF-κB were blocked (p < 0.05). In conclusion, the ß2-AR/cAMP/PKA pathway promoted apoptosis and affected the development of the uterus through the ERK and NF-κB signaling pathway. The findings of this study may provide evidence for female reproduction under psychological stress.

Nitrogen-doped carbon dots from rhizobium as fluorescence probes for chlortetracycline hydrochloride.

Fluorescent nitrogen-doped carbon dots (CDs) were prepared via hydrothermal method at 190 °C for 10 h using rhizobium from soy as the carbon and nitrogen source. Their optical properties, structure, morphology, and functional groups were characterized in detail and the results showed that they possess unique excitation-dependent fluorescence behavior, with average diameter 4.5 ± 2.0 nm and good water dispersibility. Due to the overlap of the UV-vis absorbance of chlortetracycline hydrochloride (CCH) and the fluorescence excitation band of CDs, the fluorescence of the prepared CDs can be quenched by CCH selectively and sensitively. The changes of the fluorescence intensity of CDs have a good linear relationship with the concentration of CCH in a wide concentration range of 5-100 µM, with a detection limit of 0.254 µM. This present method has been successfully applied to determine the CCH in water with recovery ranging from 96.0% to 100.7%.

Effect of chronic cyclic heat stress on the intestinal morphology, oxidative status and cecal bacterial communities in broilers.

The objective of this study was to examine the effects of chronic cyclic heat stress (HS) on the intestinal morphology, oxidative stress and cecal bacterial communities of broilers. One-day-old Arbor Acres (AA) male broilers (n = 100) were acclimated for 3 weeks and then randomly allocated into two groups, normal control (NC) group (22 ± 1 °C, 24 h/day) and HS group (32 ± 1 °C, 10 h/day lasted for 2 weeks). At 35 d of age, intestinal segments (duodenum, jejunum and ileum) and cecal digesta were collected for detection. HS affected intestinal morphology, inducing epithelial cell abscission, inflammatory cell infiltration, and lamina propria edema. Compared with the NC group, HS significantly decreased (P < 0.01) villus height (VH) and the VH-to-crypt depth (CD) ratio (VCR), increased (P < 0.05) CD in the duodenum and ileum, but had no effect on the VH in the jejunum. Moreover, HS induced oxidative stress with antioxidant enzymes activity decreasing (P < 0.05) while malondialdehyde (MDA) content increasing (P < 0.05) in small intestine. Pearson's correlation analysis indicated that MDA content was negatively correlated with VH (P < 0.05). The result of 16S rRNA sequencing showed that HS exposure impacted cecal microbiota alpha diversity (phylogenetic diversity whole-tree index) and beta diversity. Based on principal coordinate analysis (PCoA) plots for weighted UniFrac metrics and unweighted pair group method with arithmetic mean (UPGMA), there were 8 discriminative features at the genus level (linear discriminant analysis score > 2). Parabacteroides, Saccharimonas, Romboutsia and Weissella were reduced, while Anaerofustis, Pseudonocardia, Rikenella and Tyzzerella were enriched in heat-stressed broilers. Collectively, these results indicated that chronic cyclic HS induced oxidative stress that caused damage to intestinal villus-crypt structures, and then altered the cecal microflora profile.

Solid state fermentation by Fomitopsis pinicola improves physicochemical and functional properties of wheat bran and the bran-containing products.

Wheat bran was solid state fermented by Fomitopsis pinicola. The results showed that the processing properties were increased by fermentation and the content of total phenol and alkylresorcinols was 5.91 and 1.55 times of the unfermented bran respectively by the 6th day. The total antioxidant capacity was 5.73 times of the unfermented sample by the 4th day. Electronic nose analysis showed that the fermented wheat bran had a special flavor. GC-MS analysis found that 4-ethyl-2-methoxy-phenol was the main flavor substance, which was sharply increased during the fermentation. Furthermore, the textural properties of the dough and bread containing fermented bran were significantly improved. The content of phytic acid in the bread was significantly decreased, while the protein, total phenol and alkylresorcinols contents were significantly increased. Results suggest that solid state fermentation by Fomitopsis pinicola is a promising way to improve wheat bran to a nutritious and flavorful cereal food ingredient.

A ratiometric fluorometric ciprofloxacin assay based on the use of riboflavin and carbon dots.

Carbon dots (CDs) were hydrothermally synthesized from selenious yeast. They were further coupled with riboflavin to form a dually emitting probe for ciprofloxacin (CIP). Under 370 nm excitation, the probe displays dual (blue and green) emissions with peaks at 443 and 510 nm. When CIP is added, the blue fluorescence of the CDs is enhanced while the green fluorescence remains unaffected. The ratio of the relative fluorescence intensities at 443 and 510 nm increases linearly in the 0.5-200 µM CIP concentration range. The fluorescent probe is selective and has a 0.13 µM detection limit. Satisfactory recoveries (97.9-101.1%) were received when the probe was used to quantify CIP in spiked water and human serum samples. Graphical abstractBlue-emissive carbon dots were prepared from selenious yeast via a hydrothermal method, and then coupled with riboflavin as a ratiometric fluorometric probe for ciprofloxacin determination.

Hypoglycemic effects of wheat bran alkyresorcinols in high-fat/high-sucrose diet and low-dose streptozotocin-induced type 2 diabetic male mice and protection of pancreatic ß cells.

In the present study, the hypoglycemic effects of wheat bran alkyresorcinols (ARs) were investigated in type 2 diabetes mellitus (T2DM) mice induced by a high-fat/high-sucrose diet (HFSD) combined with low dose streptozotocin (STZ). After the consumption of 5 mg kg-1 d-1 acarbose (positive control) and different doses of wheat bran ARs (50, 200 and 500 mg kg-1 d-1) for 4 weeks, the fasting blood glucose (FBG) levels in T2DM mice were found to be reduced significantly (p < 0.05), and the effects of 200 and 500 mg kg-1 d-1 administration were better than that of 50 mg kg-1 d-1. The results of the oral glucose tolerance test (OGTT) also showed that both acarbose and AR administration significantly increased the glucose tolerance of the T2DM mice. Then, the fasting serum insulin levels (FINS) were significantly reduced by AR treatment, and the effect of 500 mg kg-1 d-1 AR administration was better than that of 5 mg kg-1 d-1 acarbose. The profile of plasma lipids was analyzed simultaneously, and the results showed that the contents of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) were significantly decreased, while the content of high-density lipoprotein cholesterol (HDL-C) was increased significantly after 200 and 500 mg kg-1 d-1 AR treatment (p < 0.05). Furthermore, 200 and 500 mg kg-1 d-1 ARs significantly increased the content of hepatic glycogen and the activity of glucokinase (p < 0.01) in T2DM mice. The relative mRNA levels of glucose transporter 2 (GLUT2) in the liver tissue were increased markedly in 200 and 500 mg kg-1 d-1 AR treatment groups (p < 0.01), and the relative mRNA levels of glucose transporter 4 (GLUT4) in the epididymal adipose tissue were increased significantly in all AR treatment groups, especially significantly higher than acarbose (p < 0.01). Histological analyses revealed that treatment with ARs exerted a protective role on pancreatic ß-cells. The results indicated that ARs could be an effective hypoglycemic active ingredient in whole grain diets.

Real-time recombinase polymerase amplification assay for the rapid and sensitive detection of Campylobacter jejuni in food samples.

Campylobacter jejuni (C. jejuni), a foodborne pathogen, is a major contributor to human bacterial gastroenteritis worldwide and detrimental to public health. It is crucial for initiating appropriate outbreak control strategies to rapidly detect C. jejuni. As a novel isothermal gene amplification technique, recombinase polymerase amplification (RPA) has been developed for the molecular detection of diverse pathogens. In this study, we developed a real-time RPA assay so as to achieve the rapid and efficient detection of C. jejuni by targeting the hipO gene. The specificity and senstivity of real-time RPA was validated and the practical applicability of the method for the detection of C. jejuni in artificially contaminated milk and chicken breast samples was proved by comparing their reaction time, sensitivity, and efficacy with those of real-time PCR and culture-based methods. Based on the real-time RPA assay, analysis time was reduced to approximately 13 mins from 60 mins and the results were as reliable as those of the real-time PCR assay. Taken together, in terms of the detection of C. jejuni, the real-time RPA method was simple, rapid, sensitive, and reliable.