Nutrient-split feeding strategy for dialysis cultivation of Escherichia coli

Reduction in nutrient loss during dialysis cultivation of Escherichia coli on a glycerol medium was investigated. A dialysis reactor with an inner fermentation and an outer dialysis chamber was used. Aerobic condition was maintained by limiting the glycerol feed rate to an optimum value which was estimated from the oxygen requirements for glycerol oxidation and oxygen transfer capacity of the reactor. High reduction in nutrient loss was achieved by using water as the dialyzing fluid. However, osmotic movement of water from the dialysis to the fermentation chamber was observed, and the final cell concentration was low. With a nutrient-split feeding strategy (feeding glycerol directly to the fermentation chamber and dialyzing with salt solution), glycerol loss was small, there was no osmotic flux of water to the fermentation chamber, and the cell concentration was high. Both glycerol and salt loss could be avoided, and a cell concentration of 170 g/L was obtained when the dialysis process was substituted by addition of XAD adsorbents to the dialysis chamber. Application of this nutrient-split feeding strategy to cell cultivation in a stirred tank reactor, coupled with dialysis in external dialyzer modules, resulted in low cell concentrations. (c) 1993 Wiley & Sons, Inc.     

枯草杆菌二联活菌对儿童巨细胞病毒性肝炎患儿肠道菌群及预后的影响

目的 探究枯草杆菌二联活菌对儿童巨细胞病毒(CMV)性肝炎患儿肠道菌群的调节作用及其对预后的影响。 方法 选择2016年5月到2019年5月我院收治的81例重症巨细胞病毒性肝炎患儿为研究对象,按照随机数字表法分为联合组(n=41)和对照组(n=40)。对照组患儿采用常规治疗方法,联合组患儿在对照组基础上联合枯草杆菌二联活菌颗粒治疗。比较两组患儿肝功能[谷丙转氨酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)]、肠道菌群(双歧杆菌属、乳杆菌属、芽胞杆菌属、肠球菌属、肠杆菌属、拟杆菌属)、CMV DNA、CMV IgM转阴率及预后。 结果 治疗后联合组患儿肠道芽胞杆菌属、肠球菌属数量高于对照组,肠杆菌属数量低于对照组,差异均有统计学意义(均P2=1.012,P=0.314)。 结论 枯草杆菌二联活菌联合常规治疗可有效促进巨细胞病毒性肝炎患儿肠道菌群平衡,提高患儿肝功能和病毒清除率,从而改善预后。     

连续负荷量新活素治疗难治性心力衰竭患者的临床疗效观察

目的:探讨连续负荷量新活素治疗难治性心力衰竭患者的临床疗效。方法:选取我院收治的65例难治性心力衰竭患者,随机分为两组。对照组在常规治疗的基础上加用硝酸甘油注射液,观察组在常规治疗的基础上加用新活素,比较两组的疗效,治疗前和治疗后24、36、48 h的肺动脉楔压(PAWP)、肺动脉压(PAP)、血清高敏C反应蛋白(hs-CRP),脑利钠肽(BNP)水平的变化。结果:治疗后,观察组有效率为87.9%,显著高于对照组(65.6%,P0.05)。两组患者治疗后24、36、48 h的PAWP和PAP降低值逐渐增加,且观察组PAWP和PAP降低值均显著高于对照组同时点(P0.05)。两组治疗后24、36、48 h血清hs-CRP、BNP水平随着时间延长而逐渐下降(P0.05),观察组治疗后24、36、48 h的血清hs-CRP、BNP水平均低于对照组同时点(P0.05)。结论:在基础治疗之上,给予连续负荷量新活素对于治疗难治性心力衰竭可提高治疗效果,减轻心脏前负荷,降低hs-CRP,BNP水平,改善心肌重构。     

Modulation of intestinal mucin composition and mucosal morphology by dietary phytogenic inclusion level in broilers

The effect of a dietary phytogenic feed additive (PFA) inclusion level in mucin monosaccharide composition, mucosal morphometry and mucus histochemistry along the broiler intestinal tract was studied. Cobb male broilers (n = 525) were allocated into five experimental treatments that, depending on the type of addition in the basal diet (BD), were labeled as follows: C (BD based on maize–soybean meal with no other additions), E1 (80 mg PFA/kg BD), E2 (125 mg PFA/kg BD), E3 (250 mg PFA/kg of BD) and A (2.5 mg avilamycin/kg BD). Samples from duodenum, ileum and cecum of 14- and 42-day-old broilers were collected and analyzed. In 14-day-old broilers, treatments E2 and E3 had higher (P < 0.01) duodenal mannose than treatments C, E1 and A. Ileal mannose was lower (P < 0.05) in treatment C compared with PFA treatments, and ileal galactose (Gal) was higher (P < 0.01) in treatments E2 and E3 compared with C and A. Polynomial contrast analysis with respect to PFA inclusion level showed that in 14-day-old broilers there was a linear increase (P = 0.001) in duodenal mannose and a quadratic effect (P = 0.038) in duodenal N-acetyl-galactosamine with increasing PFA level. Ileal Gal and mannose increased linearly (P = 0.002 and P = 0.012, respectively) with PFA inclusion level. There were no significant differences between treatments in mucin monosaccharide molar ratios of 42-day-old broilers. However, increasing PFA inclusion level resulted in a linear decrease of ileal fucose (P = 0.021) and cecal N-acetylgalactosamine (P = 0.036). Experimental treatments did not differ (P > 0.05) regarding duodenal villus height (Vh), crypt depth (Cd) and Vh/Cd ratio, irrespective of broiler age and the intestinal segment examined. However, increasing dietary PFA inclusion level showed a pattern of linear increase of duodenal Vh/Cd ratio in 14-day-old broilers and ileal Vh in 42-day-old broilers (P = 0.039 and P = 0.039, respectively). Alcian Blue–Periodic Acid-Schiff (pH 2.5) staining of neutral and acidic mucins showed that the staining intensity of mucus layer in villi was fragment (i.e. tip, midsection and base) dependent, whereas in crypts it was dependent both on intestinal segment (i.e. duodenum, ileum and cecum) and fragment. Finally, mucus layer thickness did not differ (P > 0.05) between treatments, yet a pattern of linear increase (P < 0.05) with PFA inclusion level was observed in the duodenum of 42-day-old broilers. In conclusion, the dietary inclusion level of PFA modulated broiler intestinal mucin composition and morphology. Further studies are required to elucidate the physiological implications of such changes in host–microflora interactions.     

Probiotic effect of lactic acid bacteria in the feed on growth and survival of fry of Atlantic cod (Gadus morhua)

A growing concern for the high consumption of antibiotics in aquaculture has initiated a search for alternative methods of disease control. Improved resistance against infectious diseases can be achieved by the use of probiotics. Probiotics are live microorganisms supplemented in food or feed which give beneficial effects on the intestinal microbial balance. In the present study a dry feed containing lactic acid bacteria (Carnobacterium divergens) isolated from Atlantic cod (Gadus morhua)intestines was given to cod fry. After three weeks of feeding the fry was exposed to a virulent strain of Vibrio anguillarum. The death rate was recorded during further three weeks of feeding with lactic acid bacteria supplemented feed. A certain improvement of disease resistance was obtained, and at the end of the experiment lactic acid bacteria dominated the intestinal flora in surviving fish given feed supplemented with lactic acid bacteria. No obvious growth inhibition of V. anguillarum was observed in an in vitro mixed culture of this bacterium and the C. divergens isolated from cod intestines. This revised version was published online in August 2006 with corrections to the Cover Date.     

Nutritional composition of black soldier fly larvae feeding on agro-industrial by-products

Black soldier fly (BSF) larvae, Hermetia illucens L. (Diptera: Stratiomyidae), bio-convert organic side streams into high-quality biomass, the composition of which largely depends on the side stream used. In the present study, BSF larvae were reared on feed substrates composed of dried brewers’ spent grains, each supplemented with either water, waste brewer’s yeast, or a mixture of waste brewer’s yeast and cane molasses to obtain 12 different substrates: barley/water, barley/yeast, barley/yeast/molasses, malted barley/water, malted barley/yeast, malted barley/yeast/molasses, malted corn/water, malted corn/yeast, malted corn/yeast/molasses, sorghum-barley/water, sorghum-barley/yeast, and sorghum-barley/yeast/molasses. The crude protein, fat, ash, and mineral contents of the BSF larvae fed each feed substrate were quantified by chemical analyses. The effect of substrate, supplementation, and their interaction on crude protein, fat, and ash contents of BSF larval body composition was significant. Calcium, phosphorus, and potassium were the most abundant macrominerals in the larvae and their concentrations differed significantly among substrates. These findings provide important information to support the use of BSF larval meal as potential new source of nutrient-rich and sustainable animal feed ingredients to substitute expensive and scarce protein sources such as fishmeal and soya bean meal.     

Post-weaning feed efficiency decreased in progeny of higher milk yielding beef cows

Current trends in the beef industry focus on selecting production traits with the purpose of maximizing calf weaning weight; however, such traits may ultimately decrease overall post-weaning productivity. Therefore, the objective of this study was to evaluate the effects of actual milk yield in mature beef cows on their offspring’s dry matter intake (DMI), BW, average daily gain, feed conversion ratio (FCR) and residual feed intake (RFI) during a ~75-day backgrounding feeding trial. A period of 24-h milk production was measured with a modified weigh-suckle-weigh technique using a milking machine. After milking, cows were retrospectively classified as one of three milk yield groups: Lower (6.57±1.21 kg), Moderate (9.02±0.60 kg) or Higher (11.97±1.46 kg). Calves from Moderate and Higher milk yielding dams had greater (P<0.01) BW from day 0 until day 75 at the end of the backgrounding feeding phase; however, day 75 BW were not different (P=0.36) between Lower and Moderate calves. Body weight gain was greater (P=0.05) for Lower and Moderate calves from the day 0 BW to day 35 BW compared with Higher calves. Overall DMI was lower (P=0.03) in offspring from Lower and Moderate cows compared with their Higher milking counterparts. With the decreased DMI, FCR was lower (P=0.03) from day 0 to day 35 in calves from Lower and Moderate milk yielding dams. In addition, overall FCR was lower (P=0.02) in calves from Lower and Moderate milk yielding dams compared with calves from Higher milk yielding dams. However, calving of Lower milk yielding dams had an increased (P=0.04) efficiency from a negative RFI value compared with calves from Moderate and Higher milking dams. Results from this study suggest that increased milk production in beef cows decreases feed efficiency during a 75-day post-weaning, backgrounding period of progeny.     

一株减毒百日咳鲍特菌的构建及生物学特性分析

百日咳是传染性强、感染率高的急性呼吸道传染病,主要感染婴幼儿,是婴儿死亡的主要原因之一。百日咳鲍特菌（Bordetella pertussis）是引起百日咳的最主要病原菌。近年来世界各地多次出现百日咳暴发,迫切需研制更加有效的新型百日咳疫苗。本研究构建了一株减毒百日咳活疫苗BPTM1,利用同源重组方法敲除编码百日咳鲍特菌主要毒力因子百日咳毒素（pertussis toxin,PTX）和皮肤坏死毒素（dermonecrotic toxin,DNT）的基因,并用大肠埃希菌的同源基因置换了负责气管细胞毒素（tracheal cytotoxin,TCT）转运的基因ampG。通过聚合酶链反应验证了毒素及相关基因的敲除和置换,蛋白免疫印迹法检测表明PTX的S1亚基未表达。体外生长曲线和体内定植曲线均表明,相比于野生型百日咳鲍特菌BPMM,减毒BPTM1的生长和定植能力未受影响,其所致肺部病理效应减轻,而所诱导的百日咳鲍特菌特异性IgG、IgG1、IgG2a抗体保持高水平。本研究表明,减毒百日咳鲍特菌BPTM1有可能成为百日咳疫苗的候选疫苗。     

激光扫描共聚焦显微镜活细胞成像方法优化

激光扫描共聚焦显微镜可用于固定样品和活细胞样品的成像,近年来得到了广泛的应用。本文介绍了激光扫描共聚焦显微镜的基本原理及其在活细胞成像中的应用,并以FV10-ASW Viewer4.2软件为例,从扫描速度、分辨率、降噪、光电倍增调节、多参数协同优化、成像质量评估、图像后期处理等多个角度总结了激光扫描共聚焦活细胞成像系统的方法优化和推荐参数设置。本文的工作可以为活细胞实验提供一定参考。     

Bacterial communities in the rumen and feces of lactating Holstein dairy cows are not affected when fed reduced-fat dried distillers’ grains with solubles

Reduced-fat dried distillers’ grains with solubles (RF-DDGSs) are co-products of ethanol production and contain less fat than traditional distillers’ grains. The fat in corn is ~91% unsaturated, and it is toxic to rumen microorganisms so it could influence the composition of the rumen microbiome. It has been demonstrated that RF-DDGS is a suitable ration ingredient to support the high-producing dairy cow, and this feedstuff is a promising alternative protein source for lactating dairy cows. The current study aims to better understand the effect of RF-DDGS on the rumen and fecal bacterial composition in lactating dairy cows. Thirty-six multiparous (two or three), mid-lactation Holstein cows (BW = 680 ± 11 kg; 106 ± 27 DIM) were randomly assigned to two groups which were fed a control diet made up of corn, corn silage, and alfalfa hay supplemented with expeller soybean meal or with added RF-DDGS (20% of the DM) containing approximately 6.0% fat. Whole rumen contents (rumen fluid and digesta; esophageal tubing method) and feces (free-catch method) were collected on day 35 of the experimental period, after the 14-d acclimation period. Rumen contents and feces from each cow were used for DNA extraction. The bacterial community composition in rumen and fecal samples was assessed via the 16S rRNA gene by using the Illumina MiSeq sequencing platform. Bacteroidetes, Actinobacteria, and Firmicutes were the most abundant phyla in rumen contents. The fecal microbiota was dominated by the phyla Firmicutes and Bacteroidetes, as well as Actinobacteria and Chloroflexi. RF-DGGS increased bacterial richness, evenness, and Shannon diversity in both rumen and fecal samples and was associated with several taxa that had different abundance in treatment versus control comparisons. The RF-DGGS, however, did not significantly alter the bacterial community in the rumen or feces. In general, these findings demonstrated that dietary inclusion of RF-DDGS did not impose any serious short-term (within 30 days) health or production consequences, as would be expected. With this study, we present further evidence that inclusion of 20% (DM basis) RF-DDGS in the diet of lactating dairy cows can be done without consequence on the microbiome of the rumen.