超高效液相色谱-四极杆-飞行时间质谱法鉴定羊尿液中安乃近的代谢物

采用超高效液相色谱-四极杆-飞行时间质谱法(UPLC/Q-TOF MS),结合全信息串联质谱(MSE)和质量亏损过滤(MDF)技术,在给药后24 h内的羊尿液中检测和鉴定了安乃近的7种代谢物(M1-M7),并根据代谢物的碎片离子特征,对代谢物进行结构初步鉴定和质谱裂解途径推测。代谢物M1、 M4、 M6、 M7被鉴定为MAA、 AA、 FAA和AAA,代谢物M2、 M3及M5被首次检测并初步鉴定为MAA和AA的羟基化产物。本实验建立了羊尿液中安乃近代谢物的实用、高效检测鉴定方法,检测和初步鉴定出7种代谢产物,为确定安乃近在羊体内的代谢途径提供了基础。     

基质辅助激光解吸电离飞行时间质谱在动物源细菌耐药性监测中的研究应用

细菌鉴定是细菌耐药性监测过程中的重要工作环节之一,基质辅助激光解析电离飞行时间质谱(Matrix-assisted laser desorption ionization time-off flight mass spectrometry, MALDI-TOF MS)能够高效鉴定细菌。为了快速监测五家养殖场来源的大肠杆菌和肠球菌的临床耐药特征,本研究利用MALDI-TOF MS和微量肉汤稀释法,快速鉴定临床分离的大肠杆菌和肠球菌,并对其进行耐药表型检测。结果显示,MALDI-TOF MS实现了对临床分离菌株(31株大肠杆菌和34株肠球菌)的快速鉴定;鸡源大肠杆菌和肠球菌的耐药情况最为严重,其次为羊和牛。其中,鸡源的大肠杆菌均对四环素(100%)和氨苄西林(91.67%)耐药率最高,肠球菌对苯唑西林(62.07%)耐药率较高。研究结果表明,不同动物源细菌临床耐药性表型严重程度有所不同,与此同时,MALDI-TOF MS技术可以同时实现对动物源大肠杆菌和肠球菌的快速鉴定,值得在动物源细菌耐药性检测领域推广应用。     

Isolation and identification of antifungal and antialgal alkaloids from Haplophyllum sieversii

Bioassay-guided fractionation of the hexane/ethyl acetate/water (H/EtOAc/H2O) crude extract of the aerial parts of Haplophyllum sieversii was performed because of preliminary screening data that indicated the presence of growth inhibitory components against Colletotrichum fragariae, Colletotrichum gloeosporioides, and Colletotrichum acutatum. Fractionation was directed using bioautographical methods resulting in the isolation of the bioactive alkaloids flindersine, anhydroevoxine, haplamine, and a lignan eudesmin. These four compounds were evaluated for activity against C. fragariae, C. gloeosporioides, C. acutatum, Botrytis cinerea, Fusarium oxysporum, and Phomopsis obscurans in a dose-response growth-inhibitory bioassay at 50.0, 100.0, and 150.0 microM. Of the four compounds tested, flindersine demonstrated the highest level of antifungal activity. Additionally, flindersine, eudesmin, and haplamine were screened against the freshwater phytoplanktons Oscillatoria perornata, Oscillatoria agardhii, Selenastrum capricornutum, and Pseudanabaena sp. (strain LW397). Haplamine demonstrated selective inhibition against the odor-producing cyanobacterium O. perornata compared to the activity against the green alga S. capricornutum, with lowest observed effect concentration values of 1.0 and 10.0 microM, respectively.     

肌肉注射盐酸头孢噻呋混悬注射液对猪安全性试验

头孢噻呋(ceftiofur)是第一个畜禽专用的第3代头孢菌素类抗生素,具有抗菌谱广、抗菌活性强、对胃酸和β-内酰胺酶较稳定、过敏反应少、毒性小、在动物体内残留低的特点[1].国内外已批准的制剂有注射用头孢噻呋钠和盐酸头孢噻呋注射液,用于肉牛、奶牛、马、羊、猪呼吸道病和1日龄鸡细菌性感染的防治[2].     

Glycoalkaloid development during greening of fresh market potatoes (Solanum tuberosum L.)

Chlorophyll and glycoalkaloid synthesis in potato (Solanum tuberosum L.) tubers occur in direct response to light. The two processes are concurrent, but independent. Color photographic indices to subjectively grade fresh market potatoes for the extent of greening were developed under lighting conditions consistent with those of retail markets. Total glycoalkaloid (TGA) and chlorophyll accumulation for four cultivars were determined over the respective greening scales, thus calibrating the scales for TGA content. On average, TGA concentrations in complete longitudinal sections of tubers (flesh samples) were highest in Dark Red Norland followed by Russet Norkotah, Yukon Gold, and White Rose. TGA concentrations of flesh samples of White Rose and Yukon Gold tubers were somewhat variable and did not increase in direct proportion to greening level and chlorophyll content, particularly at higher levels of greening. TGA concentrations in Dark Red Norland and Russet Norkotah tubers were highly correlated (P < or = 0.001) with greening level and chlorophyll concentrations. When averaged over greening levels, skin samples contained 3.4- to 6.8-fold higher concentrations of TGAs than flesh samples, depending on the cultivar. The TGA concentration in periderm samples ranged from 37 to 160 mg/100 g of dry wt. Regardless of greening level, concentrations of TGAs in the flesh samples (including attached periderm) remained within limits presumed safe for human consumption. Discrimination of greened tubers on the basis of perceived glycoalkaloid toxicity is likely unfounded for the cultivars and greening levels studied.     

Heat-induced aggregation of whey proteins: comparison of cheese WPC with acid WPC and relevance of mineral composition

Heat-induced aggregation of whey proteins in solutions made from two commercial whey protein concentrates (WPCs), one derived from mineral acid whey (acid WPC) and the other from cheese whey (cheese WPC), was studied using polyacrylamide gel electrophoresis (PAGE), size exclusion chromatography (SEC), and transmission electron microscopy (TEM). Heat treatment (75 degrees C) of acid WPC solutions (12.0%, w/w, pH 6.9) resulted in formation of relatively small "soluble" aggregates that were predominantly disulfide-linked. By contrast, heat treatment of the cheese WPC solutions (under the same conditions) caused formation of relatively large aggregates, containing high proportions of aggregates linked by noncovalent associations. The rate of aggregation of both beta-lactoglobulin and alpha-lactalbumin at 75 degrees C, measured as the loss of native proteins by PAGE, was higher in the cheese WPC solution than in the acid WPC solution. Cross dialysis of the two WPC solutions resulted in alteration of the mineral composition of each WPC solution and reversing their heat-induced aggregation behavior. The results demonstrated that the mineral composition is very important in controlling the aggregation behavior of WPC products.     

Oxidative stability of conjugated linoleic acid isomers

Conjugated linoleic acids (CLAs) have been shown to be a strong anticarcinogen in a number of animal models. Our previous study demonstrated that CLA as a whole was extremely unstable in air. The present study was undertaken further to examine the oxidative stability of individual CLA isomers using the combination of gas-liquid chromatography (GLC) and silver ion high-performance liquid chromatography (Ag-HPLC). It was found that CLA as a whole oxidized rapidly and more than 80% was degraded within 110 h in air at 50 degrees C. Four c,c-CLA isomers were most unstable followed by four c,t-CLA isomers. In contrast, four t,t-CLA isomers were relatively stable under the same experimental conditions. Both the oxygen consumption and the GLC analysis revealed that 200 ppm jasmine green tea catechins (GTCs) exhibited protection to CLA and were even stronger than 200 ppm butylated hydroxytoluene (BHT) when added to either CLA or canola oil containing 10% CLA. The present study emphasized that oxidative unstability of CLA should not be overlooked although CLA has many biological effects.     

Pressure-induced unfolding and aggregation of the proteins in whey protein concentrate solutions

Whey protein concentrate solutions (12% w/v, pH 6.65 +/- 0.05) were pressure treated at 800 MPa for 20-120 min and then examined using size exclusion chromatography (SEC), small deformation rheology, transmission electron microscopy, and various types of one-dimensional (1D) and two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE). The pressure-treated samples showed a time-dependent loss of native whey proteins by SEC and 1D PAGE and a corresponding increase in non-native proteins and protein aggregates of different sizes. These aggregates altered the viscosity and opacity of the samples and were shown to be cross-linked by intermolecular disulfide bonds and by noncovalent interactions using 1D PAGE [alkaline (or native), sodium dodecyl sulfate (SDS), and SDS of reduced samples (SDS(R))] and 2D PAGE (native:SDS and SDS:SDS(R)). The sensitivity of the major whey proteins to pressure was in the order beta-lactoglobulin B (beta-LG B) > beta-LG A > bovine serum albumin (BSA) > alpha-lactalbumin (alpha-LA), and the large internal hydrophobic cavity of beta-LG may have been partially responsible for its sensitivity to high-pressure treatments. It seemed likely that, at 800 MPa, the formation of a beta-LG disulfide-bonded network preceded the formation of disulfide bonds between alpha-LA or BSA and beta-LG to form multiprotein aggregates, possibly because the disulfide bonds of alpha-LA and BSA are less exposed than those of beta-LG either during or after pressure treatment. It may be possible that intermolecular disulfide bond formation occurred at high pressure and that hydrophobic association became important after the high-pressure treatment.     

Fumonisin B-glucose reaction products are less toxic when fed to swine

The effects of fumonisin B-glucose reaction products in swine diets was examined. Pigs were fed diets containing 528 micromol of total fumonisin B/kg (FB), 528 micromol of total FB-glucose adducts/kg (FB-G, 122 micromol of unreacted FB/kg), or 0 micromol of total FB/kg for 15 days to test the efficacy of the FB-G reaction products in detoxifying FB. Weight gain in FB pigs was lower than in FB-G or controls, which was correlated with feed intake reduction in FB pigs. Serum aspartate aminotransferase, gamma-glutamyltransferase, and total bilirubin in FB pigs were higher than in FB-G or control pigs. Serum sphinganine/shingosine ratios in FB pigs were higher than in FB-G or control pigs. Microscopic examination of tissues from FB pigs showed generalized liver necrosis and apoptosis with marked cellular pleomorphism and disorganized hepatic cords. The liver and kidneys in the FB-G group appeared to be normal. Tissues of controls were free of lesions. Results suggest that dietary FB-G products are less toxic to swine and may provide an detoxification approach in instances of widespread FB grain contamination (p < 0.05).