Structure and possible function of <Emphasis Type="Italic">N</Emphasis>-glycans of an invertebrate C-type lectin from the acorn barnacle <Emphasis Type="Italic">megabalanus rosa</Emphasis>

ABSTRACT:   A C-type lectin (BRA-2) isolated from the acorn barnacle Megabalanus rosa , which was a glycoprotein having an N -linked sugar chain, was deglycosylated by N -glycopeptidase F. The structure of the released sugar chains was determined by a 2-D mapping method after derivatization with a fluorescent reagent, 2-aminopyridine, to be Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc and Manα1-6(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc. The structures were confirmed by matrix-assisted laser desorption ionization mass spectrometry and a comparison with authentic sugar chains by high-pressure liquid chromatography. Various properties of BRA-2 were examined before and after deglycosylation. The susceptibility of BRA-2 to protease digestion was increased by deglycosylation. However, the inhibitory activity toward calcium carbonate crystallization as well as the hemagglutinating activity of deglycosylated BRA-2 was significantly decreased. These results suggest that the sugar chains of BRA-2 are important to both its structural stability and its function.     

Application of unsteady flow analysis in designing a multiple outlets sprinkler irrigation system

A simplified method has been developed for designing and calculating sprinkling systems by cutting lines across laterals at points next to the first sprinkler with a view to divide the system into two separate parts; field and basic. Equations of lateral pressure heads were used to express the relationship between inlet pressure head and velocity of the flow pumped to laterals on the field part. The above equations were then used to set up calculating conditions for the Basic part by unsteady flow method in order to define head and discharge at downstream end of the pipe in the Basic part. The forward step method was used for determining distributions of the pressure head and the emitter discharge along each lateral. A calculating schema on computer has been carried out in order to apply the method to any sprinkling system. The procedure of analysis and calculation showed that this method is generally accurate, simple and rapid.     

Time course of changes in antioxidant activity of milk from dairy cows fed a trehalose‐supplemented diet

This study was to investigate the time course of changes to the antioxidant activity of milk from cows fed a trehalose‐supplemented diet, and to determine possible underlying mechanisms for observed changes. Six Holstein cows were used, and subjected to two experimental feeding periods consisting of a 1% trehalose‐supplemented diet for 10 days, followed by a basal diet only (no trehalose) for 10 days. 1,1‐Diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging activities in milk were gradually increased during the trehalose supplementation period and were highest at the end of the second period. However, trehalose was not detected in the milk and plasma of dairy cows fed a diet supplemented with trehalose for 10 days, indicating that the increased antioxidant activity in the milk of trehalose‐fed cows is not due to the direct transfer of trehalose to the milk. Plasma DPPH activities exhibited a similar time course to that seen for milk. Relative superoxide dismutase (SOD) activities in the rumen were higher 3 days after the end of trehalose supplementation than at any other time during the experimental periods. These results suggested that the improved antioxidant activity in milk and plasma of cows fed a trehalose‐supplemented diet was due to improved ruminal relative SOD activity.     

The mcrA gene and 16S rRNA gene in the phylogenetic analysis of methanogens in the rumen of faunated and unfaunated cattle

The influence of rumen protozoa on the composition of rumen methanogens was studied by using seven growing Holstein cattle divided into two groups: four faunated and three unfaunated. 16S ribosomal RNA gene (rDNA) and methyl coenzyme‐M reductase (MCR) α subunit (mcrA) gene clonal libraries were constructed. The results of each analysis showed that Methanobacteriales was dominant in the rumen of both groups. By mcrA gene analysis, 22.1% of unfaunated clones were classified into unfaunated group 1, which was not detected from faunated cattle. The 16S rRNA gene analysis showed that the number of operational taxonomic units was higher in unfaunated than faunated cattle, suggesting the diversity of methanogens tended to be higher by the removal of protozoa. The results of the LIBSHUFF program indicated that the 16S rRNA gene and mcrA gene clone libraries for the faunated group differed from those for the unfaunated group (P = 0.001). It was suggested that the presence of protozoa strongly affected the composition of rumen methanogens.     

瘤胃非降解蛋白质对高产奶牛瘤胃发酵、血液代谢产物、产乳量和乳成分的影响

选用10头经产荷斯坦种乳牛分为对照和试验两组,以二乘反转法,研究了增加日粮中非降解蛋白质(RUP)的含量对高产奶牛瘤胃发醇、产乳量和乳成分的影响。两组日粮基本相同,试验组日粮是用加热大豆粕替代了对照组日粮中的生大豆粕,使RUP比例提高3.9%。结果表明,RUP比例的提高对瘤胃液氨氮浓度、纤毛虫种类及其数量和挥发性脂肪酸浓度及其组成无显著影响(P>0.1)。试验组牛血浆葡萄糖浓度有下降趋势(P<0.1),尿素氮、总游离氨基酸和必需氨基酸浓度两组间均无明显差异(P>0.1)。虽然两组乳牛干物质采食量无明显差异,但是试验组产乳量、4%脂肪校正乳量、乳糖和无脂固形物生产量显著增加(P<0.05),分别比对照组提高7.1%(2.3kg/d)、6.1%(1.9kg/d)、5.8%(80g/d)和6.8%(180g/d);乳蛋白质和乳脂肪生产量稍有提高,但无明显差异(P>0.1)。试验组比对照组乳蛋白质率低0.09%单位(P<0.1),乳脂率、乳糖率和无脂固形物率低0.06%～0.08%单位(P>0.1),乳脂中脂肪酸比例无明显变化。以上结果显示:提高高产奶牛日粮中RUP比例可增加产奶量,但是同时要想改善乳蛋白质含量为主的乳成分,需要平衡地改善能量、蛋白质乃至葡萄糖营养的供给状况。     

Comparison of digestibility, passage rate and rumen fermentation between sika deer (Cervus nippon) and cattle fed alfalfa hay cubes

Five female sika deer and three male Holstein cattle were offered alfalfa hay cubes at 2% (deer) and 2.5% (cattle) of bodyweight, respectively. The passage rate through the digestive tract, digestibility and rumen fermentation of the animals were determined. The rate of ruminal passage was higher and the total mean retention time in the digestive tract was shorter in deer than in cattle. In addition, the rate of post‐ruminal passage in deer was lower. The digestibilities of dry matter, organic matter, crude protein and fiber in deer were significantly lower than in cattle (P < 0.05). The concentration of total volatile fatty acids in the rumen was significantly higher in deer than in cattle. The molar percentage of acetic acid was lower and that of valeric acid was higher in deer (P < 0.05). The number of protozoa was somewhat higher in deer. These results suggested that the lower digestibility in deer might be a result of the shorter retention time in the digestive tract.     

Increase of ruminal fiber digestion by cellobiose and a twin strain of Saccharomyces cerevisiae live cells in vitro

This experiment was designed to investigate the effects of different concentrations of cellobiose (CB) or a twin strain of Saccharomyces cerevisiae live cells (YST) (20, 40 and 60 mg/60 mL), and CB + YST (60 + 20, 60 + 40, 60 + 60 mg/60 mL) on mixed ruminal microorganism fermentation in vitro. Ruminal fluid was collected from a cow, mixed with phosphate buffer (1:2) and incubated (60 mL) anaerobically at 38°C for 24 h with or without supplement plus 400 mg (dry matter [DM] basis) substrate (hay plus concentrate, 1.5:1). The medium pH numerically decreased with CB and CB + YST, but was unchanged with YST. The total volatile fatty acid and proportion of propionate increased (P < 0.05) in all cases. The proportion of acetate decreased (P < 0.05) with CB and CB + YST, but increased (P < 0.05) with YST and that of butyrate increased (P < 0.05) with CB and CB + YST, but decreased (P < 0.05) with YST. Ammonia‐N decreased (P < 0.05) with CB and CB + YST, but was unchanged with YST. The number of protozoa was unchanged, and that of cellulolytic bacteria increased (P < 0.05) in all cases. Total gas production increased (P < 0.05) in all cases. Methane decreased, hydrogen was unchanged by YST and both gases were unchanged by CB and CB + YST. The in vitro disappearance of DM and neutral detergent fiber increased (P < 0.05) by 11.2% and 8.9%, 9% and 8.5%, and 12.1% and 10.2% in the case of CB, YST and CB + YST, respectively. Therefore, the dietary supplementation of CB and/or YST may improve ruminal fermentation and digestibility.     

Comparative observations of the growth changes of the histochemical properties and collagen architecture of the iliotibialis lateralis muscle from Silky, layer and meat type cockerels

Growth‐related changes in the histochemical property and collagen architecture of the iliotibialis lateralis muscle were compared among Silky, layer and meat cockerels. Histochemical and immunohistochemical methods were employed to observe the collagen architecture. The total intramuscular collagen was also determined. The muscle consisted of type IIA, IIB and IIC myofibers, of which type IIB occurred at the highest frequency. The diameter of type IIB myofibers in each week was largest in the layer, followed by the meat, and was smallest in the Silky. The total amount of collagen reached 3.38 mg/g in the meat bird, 3.03 mg/g in the layer and 2.71 mg/g in the Silky by 30 weeks of age, respectively. In the perimysium, the collagen bundles increased in size and density of fibrils with growth. At 30 weeks of age the layer had compact collagen platelets while the Silky had loose collagen bundles. In the meat bird, the collagen bundles were moderately compact. The endomysial collagen network had a large mesh size at 1 week and thereafter accumulated many collagen fibrils to form a felt‐like fabric of fibrils at 30 weeks of age. From these results it appears that growth‐related changes in the iliotibialis lateralis muscle are not necessarily causally affected by the different growth rates of chicken breeds.     

Different patterns of vasoactive intestinal polypeptide (VIP)-immunoreactive and acetylcholinesterase (AChE)-positive innervation in the internal carotid artery and cerebral arterial tree of the quail

The innervation pattern of vasoactive intestinal polypeptide-immunoreactive (VIP-IR) nerves in the quail internal carotid artery (ICA) and cerebral arterial tree was investigated and compared with that of acetylcholinesterase-positive (AChE-P) nerves. The supply of VIP-IR nerves to the two arterial systems was distinctly richer than that of AChE-P nerves. It was focused mainly on the walls from the distal ICA to the caudal half of the anterior ramus (AR) through the cerebral carotid artery (CCA). Indeed, double staining clearly showed that numerous VIP+/AChE-axons were distributed over these arterial regions where VIP+/AChE+ or AChE+/VIP- axons were sporadic or often lacking. The finding that nerve bundles accompanying the ICA within the carotid canal contained abundant VIP+/AChE- nerve cells suggests that cerebrovascular VIP-IR nerves in the quail have their major source at these neurons and enter the cranial cavity through the CCA. Another significant finding was that a small number of nerve cells, which were mostly stained for AChE alone and occasionally for VIP alone or both, occurred in the major arteries located more rostral than the middle AR. Thus, the quail cerebral arterial tree, at least the rostral segment of the anterior circulation, is multiply innervated by these three distinct categories of the extracranial and intracranial VIP-IR and AChE-P neurons.