Isolation of Intact Chloroplasts from Spinach Leaf by Centrifugation in Gradients of the Modified Silica “Percoll”

The isolation of the photosynthetically competent chloroplast preparations was undertaken by means of the density gradient centrifugation on the modified silica sol “Percoll.” A clear separation of the intact chloroplast sustaining the high photosynthetic activities (light dependent CO₂ fixation ca. 130μmol/mg Chl·hr) was established. The contamination of mitochondria and peroxisomes was estimated to be less than 3% by measuring the activities of their marker enzymes. The chloroplasts were proved to be free from endoplasmic reticulum and cytosol. The photosynthetic CO₂ fixation of the isolated chloroplast preparations was saturated by illumination of the light intensity of 20,000 Lux (12 mW/cm², 400~750 nm).     

A hierarchical nest survival model integrating incomplete temporally varying covariates

Nest success is a critical determinant of the dynamics of avian populations, and nest survival modeling has played a key role in advancing avian ecology and management. Beginning with the development of daily nest survival models, and proceeding through subsequent extensions, the capacity for modeling the effects of hypothesized factors on nest survival has expanded greatly. We extend nest survival models further by introducing an approach to deal with incompletely observed, temporally varying covariates using a hierarchical model. Hierarchical modeling offers a way to separate process and observational components of demographic models to obtain estimates of the parameters of primary interest, and to evaluate structural effects of ecological and management interest. We built a hierarchical model for daily nest survival to analyze nest data from reintroduced whooping cranes (Grus americana) in the Eastern Migratory Population. This reintroduction effort has been beset by poor reproduction, apparently due primarily to nest abandonment by breeding birds. We used the model to assess support for the hypothesis that nest abandonment is caused by harassment from biting insects. We obtained indices of blood‐feeding insect populations based on the spatially interpolated counts of insects captured in carbon dioxide traps. However, insect trapping was not conducted daily, and so we had incomplete information on a temporally variable covariate of interest. We therefore supplemented our nest survival model with a parallel model for estimating the values of the missing insect covariates. We used Bayesian model selection to identify the best predictors of daily nest survival. Our results suggest that the black fly Simulium annulus may be negatively affecting nest survival of reintroduced whooping cranes, with decreasing nest survival as abundance of S. annulus increases. The modeling framework we have developed will be applied in the future to a larger data set to evaluate the biting‐insect hypothesis and other hypotheses for nesting failure in this reintroduced population; resulting inferences will support ongoing efforts to manage this population via an adaptive management approach. Wider application of our approach offers promise for modeling the effects of other temporally varying, but imperfectly observed covariates on nest survival, including the possibility of modeling temporally varying covariates collected from incubating adults.     

Über bakteriostatische Effekte der Darmschleimhaut von Küken mit und ohne Zulage verschiedener Wachstumsstimulatoren zur Nahrung

The supplementation of conjugated linoleic acids (CLA) to the rations of dairy cows represents an opportunity to reduce the content of milk fat. Therefore, CLA have the potential beneficial effect of reducing energy requirements of the early lactating cow. The present study aimed at the examination of long-term and post-treatment effects of dietary CLA intake on performance, variables of energy metabolism-like plasma levels of non esterified fatty acids (NEFA) and ß-hydroxybutyrate (BHB), and fatty acid profile in milk fat. Forty-six pregnant German Holstein cows were assigned to one of three dietary treatments: (1) 100 g/d of control fat supplement (CON), (2) 50 g/d of control fat supplement and 50 g/d of CLA supplement (CLA-1) and (3) 100 g/d of CLA supplement (CLA-2). The lipid-encapsulated CLA supplement consisted of approximately 10% of trans-10, cis-12 CLA and cis-9, trans-11 CLA each. The experiment started 1 d after calving and continued for about 38 weeks, divided into a supplementation (26 weeks) and a depletion period (12 weeks). Over the first 7 weeks of treatment, 11 and 16% reductions in dry matter intake compared to control were observed for the cows fed CLA-1 and CLA-2 supplements respectively. Consequently, the calculated energy balance for these two CLA groups was lower compared to the control. Plasma levels of NEFA and BHB remained unaffected. Later in lactation the highest CLA supplementation resulted in a reduction of milk fat content of 0.7%. However, no reduction in milk fat yield, and accordingly no milk fat depression (MFD), could be shown. The trans-10, cis-12 CLA in milk fat increased with increasing dietary CLA supplementation in a dose-dependent manner. The proportion of C₁₆ in milk fat was decreased by the highest CLA supplementation. With the exception of an increase in plasma glucose level in the CLA-2 group, no post-treatment effects were observed. Overall, under the conditions of the present study no improvement in the calculated energy balance by CLA supplementation could be shown for the entire evaluation period.     

Influence of hay particle size at different concentrate and feeding levels on digestive processes and feed intake in ruminants. 2. passage,digestibility and feed intake

In order to study the main effects of particle size, three ruminally fistulated cows (550 to 580 kg BW) were fed a constant low concentrate level (3.56 kg DM/d, 20% of total DMI) and a fibre‐rich hay (approximately 60% NDF in DM) in long (28.7 mm), chopped (9.2 mm) and fine ground (2.9 mm) form in a 3 x 3 Latin square design. In another three factorial experiment with 8 wethers (4 animals were ruminally fistulated, mean BW = 68 kg) the main effects and interactions of the above mentioned hay particle sizes at two concentrate levels (10.4 to 13.3% and 29.5 to 40.1% of DMI, resp.) and two intake levels (restricted and ad libitum) were investigated. In comparison to long hay (28.7 mm), feeding of chopped hay (9.2 mm) at low concentrate levels, increased not only the hay intake (7% in dairy cows and 13% in sheep) but also the intake of digestible organic matter (12% in dairy cows and 32% in sheep), due to an increase in the apparent digestibility of OM by 3.8% in dairy cows and 8.2% in sheep. Ad libitum feeding of fine ground hay in combination with low concentrate amount in the ration increased the passage rate in the hindgut and consequently the hay intake, but not the intake of DOM, due to a significant depression of digestibility, especially of fibre fractions (4 to 7% in dairy cows and 4.5 to 14% in sheep), in comparison to 28.7 and 9.2 mm hay particle sizes. The digestibility decreased significantly with restricted feeding of fine ground hay in sheep only in comparison to 9.2 mm particle size. A threefold increase of concentrate amount levelled out all effects of the particle size reduction. The effect of particle size was more pronounced in sheep than in dairy cows.     

Effects of rotational and continuous grazing on herbage quality,feed intake and performance of sheep on a semi-arid grassland steppe

Compared to continuous grazing (CG), rotational grazing (RG) increases herbage production and thereby the resilience of grasslands to intensive grazing. Results on feed intake and animal performance, however, are contradictory. Hence, the objective of the study was to determine the effects of RG and CG on herbage mass, digestibility of ingested organic matter (dOM), organic matter intake (OMI) and live weight gain (LWG) of sheep in the Inner Mongolian steppe, China. During June–September 2005–2008, two 2-ha plots were used for each grazing system. In RG, plots were divided into four 0.5-ha paddocks that were grazed for 10 days each at a moderate stocking rate. Instead, CG sheep grazed the whole plots throughout the entire grazing season. At the beginning of every month, dOM was estimated from faecal crude protein concentration. Faeces excretion was determined using titanium dioxide in six sheep per plot. The animals were weighed every month to determine their LWG. Across the years, herbage mass did not differ between systems (p = 0.820). However, dOM, OMI and LWG were lower in RG than in CG (p ≤ 0.005). Thus, our study showed that RG does not improve herbage growth, feed intake and performance of sheep and suggests that stocking rates rather than management system determine the ecological sustainability of pastoral livestock systems in semi-arid environments.     

Influence of weaning regimen on intake,growth characteristics and plasma blood metabolites in male buffalo calves

Experiment was conducted to evaluate the effect of weaning age on growth performance, feed intake, feed efficiency (FE) and blood metabolites in Nili-Ravi male buffalo (Bubalus bubalis) calves. Twenty-four male buffalo calves were assigned to one of the three treatment groups: continuous milk feeding (CMF), limited milk feeding (LMF) and early weaning (EW), and weaned off milk at 12, 10 and 8 weeks of age, respectively. For the first 3 days after birth, calves in all three treatments were fed colostrum, and were then moved to individual milk feeding at 10% of BW for the next 6 weeks. Thereafter, the provision of milk to the CMF group was gradually tapered to zero through week 12, using week 6 intakes as a base. The LMF calves were fed milk at 7.5%, 5.0%, 3.5%, and 1.5% of BW during weeks 7 to 10, respectively. Lastly, calves in the EW group were fed milk at 5.0% and 2.5% of BW at weeks 7 and 8, respectively. Calf starter (CS) feed was also provided ad libitum from weeks 2 to 12 and individual intakes were recorded on a daily basis. Blood samples were taken from weeks 6 to 12, on a weekly basis; whereas, the BW, heart girth, withers height and hip width were measured at the start of experiment and later on a weekly basis. Weight gain, average daily gain, and body measurements were the same across all three groups. Milk intake was lower (P < 0.05), whereas CS intake was greater (P < 0.05) in the EW calves compared with the other treatment groups. Dry matter intake was greater (P < 0.05) in the EW and LMF calves compared with the CMF calves. The FE was greater (P < 0.05) in the CMF calves compared with the LMF and EW treatment groups. Blood glucose concentration was similar among the treatments; however, blood urea nitrogen was greater (P < 0.05) in the EW calves compared with the CMF and LMF groups. Plasma concentration of non-esterified fatty acids was higher (P < 0.05) in the EW calves compared with the CMF calves. In light of these results, it is evident that buffalo calves can be successfully weaned as early as 8 weeks of age without negatively affecting their growth performance.     

The effects of and interactions between the maturity of grass silage and concentrate starch source when offered as total mixed rations on the performance of dairy cows

A 2 × 2 factorial feeding experiment was conducted to examine the effects of varying the maturity level of the grass used to prepare silage and the nature of concentrate starch source and their interactions on dry matter intake (DMI), diet digestibility, energy corrected milk (ECM) production and milk composition in dairy cows. Twenty-eight multiparous Swedish Red dairy cows, 133 ± 45 days in milk (DIM), with an average milk yield of 30 ± 4 kg/day and a live weight of 624 ± 69 kg were blocked by DIM and randomly assigned to seven replicated balanced 4 × 4 Latin squares with four 21-day experimental periods. The experimental diets consisted of four total mixed rations (TMR) consisting of early-cut grass silage (EGS) supplemented with either barley- or maize-based concentrate and late-cut grass silage (LGS) supplemented with either barley- or maize-based concentrate. All TMR contained identical proportions of forage (51%) and concentrate (49%). Total tract digestibility was estimated by determining indigestible NDF (iNDF) concentrations in feeds and faeces and using iNDF as an internal marker. The feeds’ ruminal degradation parameters were determined using both in situ (nylon bag) and in vitro (gas production (GP)) techniques. Cows offered diets containing EGS had greater (P < 0.001) daily dry matter (DM) intakes, ECM yields and total tract digestibilities for DM and organic matter (OM), but these were not affected by the nature of the concentrate starch source. No interaction between the maturity of the silage and the nature of the concentrate starch source was observed for DMI, diet digestibility or ECM yield. Both grass silages and concentrates had similar rates of ruminal degradation of NDF when measured in situ. The in situ DM (P < 0.001) and starch (P = 0.001) degradation rates of barley-based concentrate were greater than those for maize-based concentrate. In vitro OM GP rates and extents were similar for both concentrate feeds. The results showed that diets containing EGS offered better animal performance and diet digestibility than diets containing LGS. The concentrate starch source did not affect animal performance, but total NDF digestibility was better with diet containing barley- than maize-based concentrate.     

铜绿假单胞菌cntRLMN操纵子介导锌离子摄取的功能鉴定

【目的】本研究以铜绿假单胞菌PAO1 (Pseudomonas aeruginosa PAO1,菌种编号ATCC15692)为对象,研究cntRLMN在锌离子摄取中的功能。【方法】在ΔznuBC的基础上,以同源重组的方法构建了cntRLMN的各种突变菌株,通过质粒接合转移的方法构建其互补菌株及lacZ转录融合报告菌株,运用β-半乳糖苷酶酶活检测研究了Zur蛋白对cntRLMN的转录调控,凝胶阻滞实验(EMSA)检验Zur蛋白与cnt启动子及cnt启动子的突变片段的体外结合,并进一步通过生长曲线分析对cntRLMN中cntR、cntL、cntN等基因的锌离子摄取功能进行了分析和鉴定。最终,通过构建大蜡螟幼虫的侵染模型来研究cntRLMN对铜绿假单胞菌毒力发挥的影响。【结果】lacZ转录融合的酶活分析显示cntRLMN受Zur蛋白的负调控,其表达以Zur蛋白依赖的方式受锌离子饥饿的诱导;EMSA实验的结果显示cntRLMN的启动子可以与His-Zur结合形成DNA-蛋白质复合体,结合位点为GCGTTATAGTATATCAT;生长曲线和大蜡螟幼虫侵染实验的分析结果显示ZnuBC和CntRLMN的功能存在互补性,仅znuBC和cntRLMN双缺失突变时菌株在限锌培养条件下的生长和对大蜡螟幼虫的毒性才受到显著抑制,说明CntRLMN代表另一种独立的锌离子摄取系统。【结论】cntRLMN是受Zur直接负调控的另一种独立的铜绿假单胞菌锌离子摄取系统,对铜绿假单胞菌毒力的发挥起重要作用。     

Associations between feed efficiency,sexual maturity and fertility-related measures in young beef bulls

The beef industry has emphasized the improvement of feed utilization, as measured by modeling feed intake through performance traits to calculate residual feed intake (RFI). Evidence supports an inverse relationship between feed efficiency and reproductive function. The objective of this study was to determine the relationship of reproductive assessments and RFI unadjusted (RFI_Koch) or adjusted for body composition (RFI_us) and the relationship among fertility-related parameters. In total, 34 crossbred bulls were housed together for 112 days of performance evaluation, followed by assessment of scrotum IR imaging, scrotal circumference, testes ultrasonography and semen quality parameters at 377±33.4 days of age. Bulls were slaughtered at 389±34.0 days of age, and analyses of carcass composition, biometrics and histomorphometry of the testis and epididymis were conducted. Bulls were grouped into two subpopulations based on divergence of RFI, and within each RFI model either by including 50% of the population (Halves, high and low RFI, n=17) or 20.6% extremes of the population (Tails, high and low RFI, n=7). The means of productive performance and fertility-related measures were compared through these categories. Pearson’s correlation was calculated among fertility-related measures. In the Halves subpopulation of the RFI_us, sperm of low-RFI bulls had decreased progressive motility (47.30% v. 59.90%) and higher abundance of tail abnormalities (4.30% v. 1.80%) than that of high-RFI bulls. In the Tails subpopulation of the RFI_Koch, low RFI displayed less variation in the scrotum surface temperature (0.62°C v. 1.16°C), decreased testis echogenicity (175.50 v 198.00 pixels) and larger (60.90 v. 56.80 mm²) but less-developed seminiferous tubules than high-RFI bulls. The evaluation of fertility-related parameters indicated that a higher percentage of immature seminiferous tubules was correlated with occurrence of sperm with distal droplets (r=0.59), a larger temperature variation at the top of the scrotum was correlated with improved sperm progressive motility (r=0.38), a lower occurrence of sperm loose head abnormalities was correlated with larger temperature variation at the lower part of the scrotum (r=−0.43), and a lower minimum testis echogenicity (r=−0.59) and smaller scrotal circumference (r=0.72) were correlated with age. The adjustment for body composition (RFI determination) enabled distinct biological inferences about reproduction and feed efficiency when compared with the non-adjusted model. However, both RFI models and the correlation analysis supported the hypothesis that feed-efficient bulls have features of delayed sexual maturity. Overall, the assessment of fertility-related measurements is important to avoid the improvement of feed efficiency at the expense of reproductive function in young bulls.     

Effects of ewes grazing sulla or ryegrass pasture for different daily durations on forage intake,milk production and fatty acid composition of cheese

Sulla (Sulla coronarium L.) forage is valued for its positive impact on ruminant production, in part due to its moderate content of condensed tannin (CT). The duration of daily grazing is a factor affecting the feed intake and milk production of ewes. In this study, the effects of grazing sulla pasture compared with annual ryegrass, and the extension of grazing from 8 to 22 h/day, were evaluated with regard to ewe forage intake and milk production, as well as the physicochemical properties and fatty acid (FA) composition of cheese. During 42 days in the spring, 28 ewes of the Comisana breed were divided into four groups (S8, S22, R8 and R22) that grazed sulla (S) or ryegrass (R) for 8 (0800 to 1600 h) or 22 h/day, and received no feeding supplement. In six cheese-making sessions, cheeses were manufactured from the 48 h bulk milk of each group. Compared with ewes grazing ryegrass, those grazing sulla had higher dry matter (DM) intake, intake rate and milk yield, and produced milk that was lower in fat and higher in casein. Ewes grazing for 22 h spent more time eating, which reduced the intake rate, increased DM and nutrient intake and milk yield, and reduced milk fat. Due to the ability of CT to inhibit the complete ruminal biohydrogenation of polyunsaturated fatty acids (PUFA), the FA composition of sulla cheese was more beneficial for consumer health compared with ryegrass cheese, having lower levels of saturated fatty acids and higher levels of PUFA and n-3 FA. The FA profile of S8 cheese was better than that of S22 cheese, as it was higher in branched-chain FA, monounsaturated FA, PUFA, rumenic acid (c9,t11-C18:2), and had a greater health-promoting index. The effect of short grazing time on sulla was attributed to major inhibition of PUFA biohydrogenating ruminal bacteria, presumably stimulated by the higher accumulation of sulla CT in the rumen, which is related to a higher intake rate over a shorter eating time. Thus, grazing sulla improved the performance of ewes, thereby increasing, especially with short grazing time, the nutritional properties of cheese fat.