Rapid, accurate and visual point-of-care testing (POCT) methods for pathogenic bacteria detection are essential for avoiding foodborne diseases caused by pathogens or their toxins. In this study, we proposed a rapid and visual detection method that we named “Cas12aVIP”. By combining recombinase polymerase amplification (RPA), a CRISPR/Cas12a system and a cationic-conjugated polythiophene derivative (poly[3-(3′-N,N,N-triethylamino-1′-propyloxy)-4-methyl-2,5-thiophene hydrochloride] (PMNT) mixed with single-stranded DNA (ssDNA)), the solution turned red in the absence of the target DNA based on conformational modifications of the conjugated backbone of PMNT, whereas it displayed yellow, thus realizing the colorimetric detection of DNA. The Cas12aVIP method yielded high specificity and no interference from other nontargeted bacteria. The detection was accomplished in 40 min and the signal could be observed by the naked eye under natural light, presenting great potential for a variety of rapid nucleic acid detection applications without requiring technical expertise or ancillary equipment.
In order to seek high profit, businesses mix beef and mutton with cheap meat, such as duck, pork, and chicken. Five pairs of primers were designed for quintuple droplet digital PCR (qddPCR) of specific genomic regions from five selected species and specificity and amplification efficiency were determined. The mixed DNA template with an equal copy number was used for detecting the accuracy and limit of multiplex PCR. The results showed that the primers and probes of the five selected species had good specificity with the minimum number of detection copies: 0.15 copies/µL beef (Bos taurus), 0.28 copies/μL duck (Anas platyrhynchos), 0.37 copies/μL pork (Sus scrofa), 0.39 copies/μL chicken (Gallus gallus), and 0.41 copies/μL mutton (Ovis aries), respectively. The five sets of primers and probes could quickly judge whether the specified meat components existed in the food commodities.
The meat of local livestock breeds often has unique qualities and flavors. In this study, three Shanghai native pig breeds (MSZ, SWT, and SHB) exhibited better meat quality traits than globalized commercial pig breeds (DLY). Subsequently, metabolomic and lipidomic differences in the longissimus dorsi (L) and gluteus (T) muscles of the Shanghai native pig breeds and DLY pig breed were compared using liquid chromatography–mass spectrometry (LC–MS). The results demonstrated that the metabolites mainly consisted of (28.16%) lipids and lipid-like molecules, and (25.87%) organic acids and their derivatives were the two most dominant groups. Hundreds of differential expression metabolites were identified in every compared group, respectively. One-way ANOVA was applied to test the significance between multiple groups. Among the 20 most abundant differential metabolites, L-carnitine was significantly different in the muscles of the four pig breeds (p-value = 7.322 × 10−11). It was significantly higher in the L and T muscles of the two indigenous black pig breeds (MSZ and SWT) than in the DLY pigs (p-value < 0.001). Similarly, lipidomic analysis revealed the PA (18:0/18:2) was significantly more abundant in the muscle of these two black breeds than that in the DLY breed (p-value < 0.001). These specific metabolites and lipids might influence the meat quality and taste properties and lead to customer preferences. Therefore, this study provided insights into the characterization of meat metabolites and lipids in Shanghai native pig breeds.
Currently, Chinese herbal feed additives (CHFA) are commonly utilized in domestic pig farms. However, their impact on the sperm quality and reproductive capacity of imported breeding boars has yet to be thoroughly explored. In this study, the effect of CHFA on the sperm quality and reproductive capacity of the imported Duroc boars was investigated. Sixteen boars were randomly divided into control group and experimental (CHFA treated) group and fed normal or CHFA-levels containing diets, respectively. The sperm quality and reproductive hormone levels were periodically tested, and the reproductive capacity with breeding sows were evaluated. The results showed that the CHFA treated group boars significantly improved sperm volume, sperm concentration, and motility and reduced the sperm abnormalities. Furthermore, the serum levels of reproductive hormone such as follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) in the CHFA treated group were significantly higher than those in the control group. Although there was no significant difference in the initial birth weight of piglets between the two groups, the CHFA treated group had a significantly higher average number of piglets born, the average number of piglets born alive, the number of piglets weaned at 28 days, and the weaning weight compared to the control group. These findings suggest that CHFA can significantly improve the sperm quality of breeding boars and enhance their reproductive hormone levels as well as the reproductive capacity, providing direct evidence for the further application of CHFA in the management of breeding boars in China.
Background: Metabolic dysfunction-associated fatty liver disease (MAFLD) is associated with an accumulation of fat in the liver, disruptions in lipid metabolism, and imbalances in the gut microbiome. Extracellular vesicles (EVs) derived from probiotics have emerged as potential mediators of host lipid metabolism effect. The precise mechanisms by which EVs derived from probiotics influence MAFLD are still not fully understood.Methods: We examined the therapeutic potential of EVs sourced from Lactobacillus salivarius SNK-6 (LsEVs) using a mouse model of MAFLD and fatty acids induced cells.Results: Oral LsEVs administration reduced weight gain, lower liver enzyme levels, and less liver fat in mice. Meanwhile, LsEVs increases the secretion of anti-inflammatory factor IL-4 in mice subjected to a high-fat diet, but inhibited the pro-inflammatory cytokine secretion in lipopolysaccharide induced gut cells. Mechanistically, LsEVs enhance liver cell mitophagy via Beclin-1 and PPAR related pathways. LsEVs also increased tight junction proteins in epithelial cells. Furthermore, LsEVs boost gut bacterial diversity in MAFLD -afflicted mice by promoting beneficial Bacteroidota and suppressing harmful Desulfovibrio.Conclusions: Our research established a foundation for the future use of LsEVs in treating MAFLD and provided novel insights into the mechanisms of lipid metabolism influenced by EVs derived from probiotics in the context of MAFLD.