单宁酸对棕色田鼠和小鼠食物选择、日食量和蛋白质消化率的影响

目的用不同浓度单宁酸（tannic acid,TA）处理后的花生喂养棕色田鼠（Lasiopodomys mandarinus）和小鼠（Mus musculus）,研究单宁酸对两种鼠食物选择倾向、食物摄入量、蛋白质消化率的影响。方法本研究的设计如下：1）选择一批棕色田鼠和小鼠,单笼饲养,自由饮水。用正常花生饲喂二周实验用棕色田鼠和小鼠,使其适应这种食物;2）随机选出棕色田鼠和小鼠各10只（雌雄各半）,单笼饲养。用经0%、5%、10%TA处理的花生饲喂供试鼠一周,观察和记录两种鼠对TA处理后花生的选择和取食特征;3）随机选出18只棕色田鼠和18只小鼠,单笼饲养,并各分为3组（每组均为6只）,按两种鼠分别命名为对照组、低单宁酸组、高单宁酸组,并分别用经0%、5%和10%TA处理的花生饲喂一周,测定两种鼠的食物摄入量;同时收集粪便,用凯氏定氮仪测定粗蛋白含量,计算蛋白质消化率。结果1）棕色田鼠和小鼠均优先选择无TA食物（P〈0.001）,二者间差异达显著性水平（P〈0.001）;2）食物中的TA降低棕色田鼠和小鼠的相对日食量（P〈0.001）和食物中蛋白质的消化率,随TA含量的升高,棕色田鼠和小鼠的相对日食量和蛋白质消化率均显著下降。结论TA降低棕色田鼠和小鼠的食物摄入量和食物中蛋白质的消化率。TA对鼠类的食物摄入量和蛋白质消化率的影响有种间差异性,小鼠对TA的适应性更强。     

构建丝状真菌大片段基因组文库载体DNA的制备

载体DNA的制备是构建大片段基因组文库的关键步骤之一,高质量载体DNA受到酶切、脱磷等因素的影响,以载体pBHYG为材料,优化了限制性内切酶胁HindⅢ酶切和小牛肠碱性磷酸酶(CLAP)脱磷的作用条件,并在T4连接酶作用下自连,通过胶回收纯化制备了可用于进一步构建大片段基因组文库的线性载体DNA。     

Performance of a generalist grasshopper on a C<Subscript>3</Subscript> and a C<Subscript>4</Subscript> grass: compensation for the effects of elevated CO<Subscript>2</Subscript> on plant nutritional quality

The increasing CO₂ concentration in Earths atmosphere is expected to cause a greater decline in the nutritional quality of C₃ than C₄ plants. As a compensatory response, herbivorous insects may increase their feeding disproportionately on C₃ plants. These hypotheses were tested by growing the grasses Lolium multiflorum C₃) and Bouteloua curtipendula C₄) at ambient (370 ppm) and elevated (740 ppm) CO₂ levels in open top chambers in the field, and comparing the growth and digestive efficiencies of the generalist grasshopper Melanoplus sanguinipes on each of the four plant × CO₂ treatment combinations. As expected, the nutritional quality of the C₃ grass declined to a greater extent than did that of the C₄ grass at elevated CO₂; protein levels declined in the C₃ grass, while levels of carbohydrates (sugar, fructan and starch) increased. However, M. sanguinipes did not significantly increase its consumption rate to compensate for the lower nutritional quality of the C₃ grass grown under elevated CO₂. Instead, these grasshoppers appear to use post-ingestive mechanisms to maintain their growth rates on the C₃ grass under elevated CO₂. Consumption rates of the C₃ and C₄ grasses were also similar, demonstrating a lack of compensatory feeding on the C₄ grass. We also examined the relative efficiencies of nutrient utilization from a C₃ and C₄ grass by M. sanguinipes to test the basis for the C₄ plant avoidance hypothesis. Contrary to this hypothesis, neither protein nor sugar was digested with a lower efficiency from the C₄ grass than from the C₃ grass. A novel finding of this study is that fructan, a potentially large carbohydrate source in C₃ grasses, is utilized by grasshoppers. Based on the higher nutrient levels in the C₃ grass and the better growth performance of M. sanguinipes on this grass at both CO₂ levels, we conclude that C₃ grasses are likely to remain better host plants than C₄ grasses in future CO₂ conditions.     

Isolation and characterization of five serine proteases with trypsin-, chymotrypsin- and elastase-like characteristics from the gut of the lugworm Arenicola marina (L.) (Polychaeta)

Summary Five proteases were isolated from the digestive fluid of the lugworm, Arenicola marina L. The enzymes (molecular weight 24.0–24.6 kDa) were classified as serine proteases. Three enzymes showed a cleavage specificity corresponding to mammalian trypsin (E.C. 3.4.21.4). One protease possessed a chymotrypsin-like cleavage pattern (E.C. 3.4.21.1), and the fifth preferred cleavage behind short-chain amino acids like an elastase (E.C. 3.4.21.36). Detailed investigations revealed differences in molecular characteristics and cleavage patterns compared to mammalian proteases, especially in the chymotrypsin- and the elastase-like enzymes.Abbreviations APNE N-acetyl-d/l-Phe -naphthyl ester - BANA N-benzoyl-d/l-Arg -naphthylamide - BAPNA N-benzoyl-d/l-Arg-4-nitroanilide - BIGGANA N-benzoyl-l-Ile-l-Glu-Gly-l-Arg-4-nitroanilide - BLPNA N-benzoyl-d/l-Lys-4-nitroanilide - BTEE N-benzoyl-l-Tyr ethyl ester - enzyme T1/T2/T3 trypsin-like enzyme - enzyme ChT chymotrypsin-like enzyme - enzyme E elastase-like enzyme - GPANA N-glutaryl-l-Phe-4-nitroanilide - MUF 4-methylumbelliferryl - MW molecular weight - PMSF phenylmethylsulphonyl fluoride - SAAPPNA N-succinyl-l-Ala-l-Ala-l-Pro-l-Phe-4-nitroanilide - SBTI soybean trypsin inhibitor - SPPNA N-succinyl-l-Phe-4-nitroanilide - TAME N-tosyl-l-Arg methyl ester - TFA trifluoracetic acid - TLCK N-tosyl-l-Lys chloromethyl ketone - TPCK N-tosyl-l-Phe chloromethyl ketone - TRIS tris(hydroxymethyl)aminomethane     

The metabolism of structured triacylglycerols

The triacylglycerol (TAG) structure in addition to the overall fatty acid profile is of importance when considering the nutritional effect of a dietary fat. This review aims at summarizing our current knowledge of the digestion, absorption, uptake, and transport of structured TAGs, with particular emphasis on the following aspects: gastric emptying, specificity of pancreatic lipase, lymphatic transport and clearance of chylomicrons, effects of lipid structure on tissue lipid compositions and the fecal loss of fats. So an overview will be provided for how the structure and fatty acid composition of TAGs affect their absorption and the distribution of the fatty acids in the body following digestion and absorption.     

In vitro analysis of the digestive enzymes amylase and alpha-glucosidase in the midguts of Locusta migratoria L. in response to the myosuppressin, SchistoFLRFamide

We have investigated the effect of the locust myosuppressin, SchistoFLRFamide, on the activity of amylase and alpha-glucosidase in the midgut of 2-week old male locusts. Total enzyme activity in the lumen contents and tissue extracts of midguts responds to SchistoFLRFamide in a dose-dependent manner that appears to vary with the feeding state of the locust and duration of exposure to the peptide. Starvation for 24h prior to assessment alters the distribution of enzyme activity between the midgut lumen contents and tissue extracts in response to SchistoFLRFamide when compared with fed locusts. Duration of exposure to SchistoFLRFamide also alters the distribution of total amylase and alpha-glucosidase activity; as duration of exposure increases, lower concentrations of SchistoFLRFamide increase total enzyme activity in the lumen contents while decreasing total enzyme activity in the tissue extracts. We suggest that the minimum amino acid sequence in SchistoFLRFamide necessary to increase both amylase and alpha-glucosidase activity is DHVFLRFamide. We have determined that two other peptides endogenous to the locust, AFIRFamide and GQERNFLRFamide, increase amylase and alpha-glucosidase activity in midgut lumen contents.     

The thermoregulatory function of diel vertical migration for a juvenile fish,Cottus extensus

Juvenile sculpin (Cottus extensus) less than 30 mm long exhibit a diel vertical migration in the limnetic zone of Bear Lake (Utah-Idaho). Using mid-water and bottom trawls we found that these fish inhabit the bottom of the lake (5° C) during the day but migrate 30–40 m into the water column at night where they reside in the metalimnion or epilimnion at temperatures near 13–16°C. Larger fish do not migrate into the water column. Stomach analyses demonstrated that the young-of-the-year fish do not migrate into the water column to feed: from July to October their diet is 70–93% benthic ostracods and copepods, and pelagic prey are rarely consumed. Furthermore, gut fullness of the sculpin increases through the daylight period and decreases through the night, reaching minimum levels just before the dawn descent. Laboratory experiments demonstrated that the diel migration would increase digestion rate from 3%/h at profundal temperatures, to 22%/h in the warmer surface water, thus allowing the fish to empty their guts overnight and permit feeding the following day. Additionally, sculpin held in a temperature and feeding regime that mimicked that experienced by migrating fish grew 300% faster than those reared at 5° C. Given the overwhelming importance of fast growth for juvenile fishes, a post-feeding thermotaxs that increases digestion may be a common phenomenon increasing growth, and affecting the distribution and bioenergetic relationships of fish.     

Wolbachia infection in Aedes aegypti mosquitoes alters blood meal excretion and delays oviposition without affecting trypsin activity

Blood feeding in Aedes aegypti is essential for reproduction, but also permits the mosquito to act as a vector for key human pathogens such as the Zika and dengue viruses. Wolbachia pipientis is an endosymbiotic bacterium that can manipulate the biology of Aedes aegypti mosquitoes, making them less competent hosts for many pathogens. Yet while Wolbachia affects other aspects of host physiology, it is unclear whether it influences physiological processes associated with blood meal digestion. To that end, we examined the effects of wMel Wolbachia infection in Ae. aegypti, on survival post-blood feeding, blood meal excretion, rate of oviposition, expression levels of key genes involved in oogenesis, and activity levels of trypsin blood digestion enzymes. We observed that wMel infection altered the rate and duration of blood meal excretion, delayed the onset of oviposition and was associated with a greater number of eggs being laid later. wMel-infected Ae. aegypti also had lower levels of key yolk protein precursor genes necessary for oogenesis. However, all of these effects occurred without a change in trypsin activity. These results suggest that Wolbachia infection may disrupt normal metabolic processes associated with blood feeding and reproduction in Ae. aegypti.     

Cytochemical and biochemical observations on the digestive tracts of digenetic trematodes. 8. Acid phosphatase activity in Schistosoma mansoni and Schistosomatium douthitti

At the light microscope level, nonspecific acid phosphatase (AcPase) (EC 3.1.3.2) and N-acetyl glucosaminidase (NAGase) (EC 3.2.1.29) activities are in the esophageal gland cells of Schistosoma mansoni and Schistosomatium douthitti and in the gastrodermis of S. mansoni. The gastrodermis of S. douthitti is negative for these two enzymes. At the electron microscope level, AcPase activity in the esophageal gland cells of both species is observed in cytoplasmic vesicles. In S. mansoni, AcPase activity is also observed associated with the infoldings of the basal plasma membranes of the esophagus and the gastrodermis. It is hypothesized that this enzyme(s) is involved with membrane transport. AcPase activity is also associated with “droplets” and vesicles in the gastrodermis of S. mansoni. It is believed that the digestion of foodstuffs in both species occurs extracellularly.     

Establishing a feed value for moulded hay

Fungal invasion in forage materials can occur during plant growth, harvest or storage. End products generated from this activity include fungal biomass, mycotoxin, and fruiting body or spore formation. Spores are predominantly produced by the thermotolerant organisms during storage. Mycotoxin production is observed for forage stored at moisture levels greater than 35%; however, there is little evidence that mycotoxins will occur in hay stored at moisture levels lower than 30%. Hay exposed to precipitation for more than 1 year has not received adequate attention regarding the potential for mycotoxin content.

Fungal biomass (a combination of viable and non-viable mycelia and spores) accumulation was evaluated relative to palatability, intake, digestibility and rumen function. Given a choice, animals will select hay with low levels of fungal biomass contamination. However, results from four studies with growing animals in which hay was offered as the sole feedstuff indicate that intake is determined by the nutrient characteristics of the feedstuff and is not depressed due to presence of mould. Fed as long hay, dry matter digestibility of moulded hay does not differ from clean hay of a similar nutrient profile. Digestibility differences for these hays may become more apparent when chopped. A possible rational may be that the actions of fungal invasion reduce integrity of the forage, causing particle size reduction to be greater when processed. Rumen function does not appear to be impaired when moulded hay is fed.