Continuous,high capacity reconstitution of nutrient media from concentrated intermediates

We designed an Integrated Media Preparation System (IMPS) for continuous, on-line preparation of cell culture media and delivery to intermediate storage vessels or directly to a bioreactor. Key components of the IMPS include: a high precision, continuous fluid mixing device; formulation-specific liquid medium concentrates; validated process controls and membrane filtration; and automated dispensing into large volume flexible plastic containers. The IMPS system is designed to produce sterile, single-strength liquid medium from common raw materials at a delivery rate of 1000–3000 liters per hour and will manufacture homogenous batches from several thousand liters to over 60,000 liters. Fortified nutrient media prepared from multi-component 50X concentrates have been demonstrated to accelerate bioreactor seed chains, increase product yield, and reduce the overall manufacturing cost of nutrient medium. A productivity matrix will analyze the fully-loaded costs and contrast alternative methods for media preparation against projected biological yield.Abbreviations IMPS Integrated Media Preparation System - 50X Nutrient fluid components formulated at fifty-fold final use concentration - 1X Nutrient fluid formulated at final, single-strength use concentration - cGMP Current Good Manufacturing Practices - SCADA Supervisory Control and Data Acquisition - PLC Process Logic Controller - LTI Life Technologies, Inc. - WFI Water for Injection - CIP Clean in place - SIP Sterilize in place - HPLC High performance liquid chromatography - DMEM Dulbecco's Modified Eagle's Medium     

Use of some emulsified plant seed oils as a safe alternative for the management of Meloidogyne incognita infecting tomato

Abstract

The aim of the present study was to formulate six different plant seed oils namely canola, cotton, flax, olive, sesame and soybean as emulsifiable concentrates. The composition of the formulation comprises at least one organic solvent, one surfactant and one plant oil. Physico-chemical properties of the formulated oils (emulsion stability test, cold stability and heat stability tests) were measured. The successfully emulsified oils were evaluated for nematicidal activity against Meloidogyne incognita infecting tomato plants under greenhouse conditions. Emulsified canola oil proved to be the most effective oil as a protectant against M. incognita infection to tomatoes followed by soybean, cotton, flax and sesame oil. In addition, employing a high rate of the tested emulsified oils gave higher activity in suppressing nematodes both in the soil and in tomato roots than using a low rate. Moreover, all tested formulated oils at both rates of application had no adverse effect on the growth of tomato plants except sesame oil which significantly decreased the shoot length when compared to the control. The prepared plant oils might be used as potential sources for sustainable eco-friendly botanical nematicides to protect plants from nematode attack.     

Untersuchungen zur Frage der Methionintoxizität

The study aimed to examine effects of supplemented conjugated linoleic acids (CLA) to periparturient cows receiving different concentrate proportions ante partum (a.p.) to investigate CLA effects on lipid mobilisation and metabolism. Compared to adapted feeding, a high-concentrate diet a.p. should induce a ketogenic metabolic situation post partum (p.p.) to better understand how CLA works. Sixty-four pregnant German Holstein cows had ad libitum access to partial mixed rations 3 weeks prior to calving until day 60 p.p. Ante partum, cows received control fat (CON) or a CLA supplement at 100 g/d, either in a low-concentrate (CON-20, CLA-20) or high-concentrate diet (CON-60, CLA-60). Post partum, concentrate proportion was adjusted, while fat supplementation continued. After day 32 p.p., half of the animals of CLA-groups changed to CON supplementation (CLA-20-CON, CLA-60-CON). A ketogenic metabolic situation p.p. was not achieved and therefore impacts of CLA could not be examined. Live weight, milk yield and composition, blood parameters remained unaffected by the treatments. Only a slightly reduced milk fat yield (not significant) was recorded for Group CLA-20. The proportion of trans-10,cis-12 (t10,c12) CLA in milk fat was significantly increased in CLA-groups compared to CON-groups. With the exception of a reversible CLA effect on milk fat in Group CLA-20, no post-treatment effects occurred. Dry matter intake (DMI) of Group CLA-60 was highest before calving, resulting in a significantly improved estimated energy balance after calving. Ante partum, net energy intakes were significantly increased in high-concentrate groups. Overall, supplemented CLA preparation did not relieve metabolism and lipid mobilisation of early lactating cows. But feeding CLA in a high-concentrate diet a.p. seems to increase DMI and thereby improve the energy balance of cows immediately after calving.     

Effects of folic acid supplementation to rations differing in the concentrate to roughage ratio on ruminal fermentation,nutrient flow at the duodenum,and on serum and milk variables of dairy cows

The present study was undertaken to determine the effects of dietary folic acid (FOL) supplementation on ruminal fermentation, duodenal nutrient flow, serum and milk variables, and on B-vitamin concentration in serum. The study was divided into two experiments: in Exp. 1 the forage to concentrate (F:C) ratio of the diet (DM basis) was 34:66 (high concentrate, HC), while in Exp. 2 the F:C ratio was 66:34 (high forage, HF). In addition, the cows received 0 or 1 g FOL/d. In Exp. 1, two German Holstein cows equipped with cannulas in the dorsal sac of the rumen and in the proximal duodenum were dry and five were lactating (186 ± 144 days in milk); in Exp. 2 four cows were dry and four were lactating (165 ± 57 days in milk). In cows fed the HC diet, FOL supplementation decreased the ruminally-fermented organic matter. Thus, less energy was available for ruminal microorganisms, which resulted in a reduced microbial crude protein flow at the duodenum. Feeding the HF diet, FOL supplementation only increased the apparent ruminal digestibility of acid detergent fibre (ADF). With the HF diet, FOL had no influence on the serum levels of glucose, non-esterified fatty acids, beta-hydroxybutyrate, urea, thiamine, riboflavin, pyridoxal-5′-phosphate, pyridoxic acid, pyridoxal, pyridoxine, pyridoxamine, pantothenic acid, nicotinamide or nicotinic acid, whereas supplementing FOL to the HC diet lowered the serum glucose and riboflavin levels. In both experiments, the supplementation of FOL had no effects on milk composition. Folic acid supplementation to both diets increased the concentrations of serum 5-methyl-tetrahydrofolate. However, no beneficial effects to dairy cows were obvious. Therefore, to achieve certain results, studies with a higher number of non-fistulated cows would be necessary.     

Effects of rumen-escape starch and coarseness of ingredients in pelleted concentrates on performance and rumen wall characteristics of rosé veal calves

The objective was to study the effect of rumen-escape starch and coarseness of ingredients in pelleted concentrates on performance, carcass quality and rumen wall characteristics in rosé veal calf production. Two alternative concentrates (Coarse and Slow) were compared with a traditional (Control) concentrate. Control was based on finely ground ingredients, whereas in Coarse, the same ingredients were coarsely ground resulting in a mean particle size before pelleting of 1.5 in Coarse and 0.6 mm in Control. Slow compared with Control and Coarse contained finely ground sorghum and corn instead of barley and wheat which increased the amount of rumen-escape starch to 59 compared with 22 g/kg in Control and Coarse. All concentrates had the same total starch (362 g/kg), NDF (168 g/kg), CP (154 g/kg) and DE (15.5 MJ/kg DM) content and a pellet diameter of 3.5 to 4 mm. Use of an ‘indicator of starch digestibility’ method gave a value of 98.6% for Control and Coarse and 91.1% for Slow (P < 0.001). A total of 57 Holstein bull calves (n = 19 per treatment) were offered one of the three concentrates ad libitum from weaning 2 1/2 months of age) to slaughter (<10 months of age). Concentrate intake was recorded individually. Barley straw was available ad libitum but intake was not recorded. Average daily gain (1.43 kg/day), concentrate conversion efficiency (3.7 kg DM concentrate/kg gain), LW at slaughter (386 kg), carcass weight (194 kg) and EUROP conformation (3.9) were not affected by type of concentrate (P > 0.05). Papillae length and shape evaluated in atrium ruminis and the cranial part of the ventral rumen sac at slaughter were not affected by type of concentrate (P > 0.05). Rumen wall characteristics showed degrees of plaque formation (i.e., papillary aggregation), hyperaemia and necrotic areas in all treatment groups, but with no general difference between type of concentrate (P > 0.05). Incidence of liver abscesses (LAs, 16%) was not affected by type of concentrate (P > 0.05). There were no differences in performance or rumen wall characteristics between liver-abscessed and non-abscessed calves. The results show a high level of production performance with the three types of pelleted concentrates and indicates that neither the more coarse ingredients nor the additional rumen-escape starch tested, when fed ad libitum, could improve rumen wall characteristics or reduce LAs of rosé veal calves.     

Development of an internally controlled PCR assay for broad range detection of bacteria in platelet concentrates

A real-time PCR assay based on the 16S rRNA gene was optimized for the detection of a broad range of bacteria in plasma and platelet concentrates (PC). A lambda phage internal control was constructed and implemented in the assay, which made it suitable for diagnostic use. Spiking studies in plasma and PCs were performed to determine the analytical sensitivity of the assay. Thirty three colony forming units (CFU)/ml of E. coli and 72 CFU/ml of Staphylococcus epidermidis could be detected in plasma, and 97 CFU/ml of S. epidermidis in PCs. The assay detected all bacteria relevant for bacterial contamination of PCs. The short turn around time of the assay made it suitable for testing PCs for bacterial contamination prior to transfusion.     

Changes in the Volatile Flavor Constituents of Citrus sinensis (L.) Osbeck During Storage

The methods of GC with fused silica capillary column, its Kovats retention index of GC and GC-MS-DS etc. We were used to study the aromatic compounds of the peel essential oil of Citrus sinensis (L.) Osbeck and aromatic concentrates of fruit flesh. 29 chemical constituents from the peels and 40 chemical constituents from the fruit flesh were identified. The trace constituents α-sinensal 0.01% and β-sinensal 0.01% are the special aroma ones for Citrus sinensis. At the same time, the changes flavor components in Citrus sinensis peels and fruit flesh during storage were also explored.     

中国福建铁观音茶、色种茶、武夷水仙茶和武夷奇种茶的香气成分研究

本文首次报道著名的中国福建乌龙茶:铁观音茶、色种茶、武夷水仙茶和武夷奇种茶的香气成分。上述茶样香气浓缩物,用 GC-MS 和 SCOT 玻璃毛细管柱气相色谱标样叠加法及红外光谱,鉴定出70种香气成分。从中找出了12种典型特征香气成分:苯乙醇、反-芳樟醇氧化物(吡喃型)、顺-芳樟醇氧化物(呋喃型)、3,7-二甲基-1,5,7-辛三烯-3-醇、苯甲醇、反-芳樟醇氧化物(呋喃型)、香叶醇、吲哚、顺-茉莉酮、茉莉酮内酯、橙花叔醇和茉莉酮酸甲酯。从而初步剖析了由不同茶树品种、不同地区以及加工风格差异所形成的香气变化规律。     

Young Salers suckled bull production: effect of diet on performance, carcass and muscle characteristics and meat quality

The aim of this work was to improve the knowledge on young suckled Salers bull production and to study the effect of forage type and concentrate level on performance, carcass and muscle characteristics as well as on meat quality. Twenty-four Salers male calves of 150 days of age were assigned to six groups: C0 (fed exclusively with hay and dam's milk and slaughtered at approximately 6 months of age), and HH (hay - high concentrate), HL (hay - low concentrate), GH (cut grass - high concentrate), GL (cut grass - low concentrate) and CP (control pasture: pasture - high concentrate) groups differing in feeds received until slaughter and slaughtered unweaned at approximately 10 months of age. Carcass weights averaged 210 kg at 10 months of age at slaughter. Average daily weight gain (ADWG) in HH and GH groups tended to be higher (P = 0.09) than in HL and GL groups (1354 v. 1248 g/day). ADWG in CP group (1542 g/day) was higher (P < 0.05) than in the other groups. Carcass weight in CP group (230 kg) tended to be higher (P < 0.1) than in HL (198 kg) and GL (200 kg) groups. Carcass muscle weight was higher (P < 0.05) in GH (155 kg) and CP groups (165 kg) than in HL (141 kg) and GL (142 kg) groups. Carcass and offal fatty tissue weights and carcass fatness did not differ between groups. Neither forage type nor concentrate level had significant effect on the area of muscle fibres or on muscle metabolic enzyme activities (namely, lactate dehydrogenase - LDH, phosphofructokinase - PFK, isocitrate dehydrogenase - ICDH, citrate synthase - CS and cytochrome-c oxidase - COX). semitendinosus muscle of CP group presented higher CS enzyme activities (8.10 μmol/min per g) than HH (5.30 μmol/min per g) and GL (4.52 μmol/min per g) groups. Neither total nor insoluble collagen content significantly differed between groups. Lipid content in rectus abdominis muscle was relatively low (average 67.5 mg/g dry matter) and was not affected by diet (P > 0.05). The ratio between n-6 and n-3 polyunsaturated fatty acid content was lower (P = 0.01) in the low-concentrate-fed than in the high-concentrate-fed group (3.95 v. 5.37, respectively). Sensory analysis noted that longissimus thoracis muscle from CP animals was more tender and juicy than that from HH and GH animals (P < 0.05).     

A new approach to estimate the in situ fractional degradation rate of organic matter and nitrogen in wheat yeast concentrates

In the classic in situ method, small particles are removed during rinsing and hence their fractional degradation rate cannot be determined. A new approach was developed to estimate the fractional degradation rate of nutrients in small particles. This approach was based on an alternative rinsing method to reduce the particulate matter loss during rinsing and on quantifying the particulate matter loss that occurs during incubation in the rumen itself. To quantify particulate matter loss during incubation, loss of small particles during the in situ incubation was studied using undegradable silica with different particle sizes. Particulate matter loss during incubation was limited to particles smaller than ~40 μm with a mean fractional particulate matter loss rate of 0.035 h⁻¹ (first experiment) and 0.073 h⁻¹ (second experiment) and an undegradable fraction of 0.001 and 0.050, respectively. In the second experiment, the fractional particulate matter loss rate after rinsing in a water bath at 50 strokes per minute (s.p.m.) (0.215 h⁻¹) and the undegradable fraction at 20 s.p.m. (0.461) were significantly larger than that upon incubation in the rumen, whereas the fractional particulate matter loss rate (0.140 and 0.087 h⁻¹, respectively) and the undegradable fraction (0.330 and 0.075, respectively) after rinsing at 30 and 40 s.p.m. did not differ with that upon rumen incubation. This new approach was applied to estimate the in situ fractional degradation rate of insoluble organic matter (OM) and insoluble nitrogen (N) in three different wheat yeast concentrates (WYC). These WYC were characterised by a high fraction of small particles and estimating their fractional degradation rate was not possible using the traditional washing machine rinsing method. The new rinsing method increased the mean non-washout fraction of OM and N in these products from 0.113 and 0.084 (washing machine method) to 0.670 and 0.782, respectively. The mean effective degradation (ED) without correction for particulate matter loss of OM and of N was 0.714 and 0.601, respectively, and significant differences were observed between the WYC products. Applying the correction for particulate matter loss reduced the mean ED of OM to 0.676 (30 s.p.m.) and 0.477 (40 s.p.m.), and reduced the mean ED of N to 0.475 (30 s.p.m.) and 0.328 (40 s.p.m.). These marked reductions in fractional degradation rate upon correction for small particulate matter loss emphasised the pronounced effect of correction for undegraded particulate matter loss on the fractional disappearance rates of OM and N in WYC products.