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     

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.     

Gut architecture,digestive constraints and feeding ecology of deposit-feeding and carnivorous polychaetes

Summary We analyze gut architectures of 42 species of marine polychaetes in terms of their anatomically distinct compartments, and quantify differences among guts in terms of ratios of body volume to gut volume, relative compartmental volumes, total gut aspect ratios and compartmental aspect ratios. We use multivariate techniques to classify these polychaetes into 4 groups: carnivores with tubular guts; deposit feeders with tubular guts; deposit feeders with 3 gut compartments; and deposit feeders with 4 or 5 gut compartments. Tubular guts, morphological expressions of plug flow, are common among deposit feeders and may allow relatively rapid ingestion rates and short throughput times. Median gut volume per unit of body volume in deposit feeders (31%) is twice that of carnivores (15%) and ranges up to 83% in one deep-sea species. Deep-sea deposit feeders tend to have relatively larger and longer guts than closely-related nearshore and shelf species. Guts of a number of deep-sea deposit feeders and nearshore and shelf deposit feeders from muddy environments are relatively longer and narrower as body size increases, suggesting that digestive diffusion limitations may be important. Gut volume scales as (body volume)¹ while ingestion rate scales as (body volume)^0.7. If diet and the chemical kinetics of digestion do not change appreciably, throughput time and thus the extent of digestion of given dietary components therefore must increase as a deposit feeder grows. Digestive processing constrainst may be most important in juveniles of species (especially those species with plug-flow guts) that are deposit feeders as adults.     

九孔鲍消化道及养殖水体中异养细菌胞外产物的分析

从汕尾健生鲍鱼场养殖水体及鲍鱼消化道中分离筛选到26株异养细菌,分析了其胞外产物,并采用API条带对其进行了种类鉴定。结果表明,就整体而言,成鲍消化道菌株产蛋白酶、淀粉酶、明胶酶和溶血能力均高于养殖水体的菌株,而产脂肪酶和卵磷脂酶菌株的比例则低于后者。无论是消化道还是养殖水体,均存在着分泌胞外产物能力极强的菌株,且大部分为鞘氨醇单胞菌。因此,在该养殖环境中,鞘氨醇单胞菌应视为一种主要的条件致病菌,同时消化道和养殖水体均应视为潜在致病菌的源泉。此外,在考虑细菌对水产经济动物的致病作用时,除了优势种类外,还应从菌群结构的角度出发,考虑整个细菌群落胞外产物的作用。     

Rumen Fermentation and Nitrogen Balance of Lambs fed Diets Containing Plant Extracts Rich in Tannins and Saponins,and Associated Emissions of Nitrogen and Methane

Tannins were added to experimental diets at levels of 1 and 2g/kg DM (hydrolysable tannins; Castanea sativa wood extract) and saponins at 2 and 30mg/kg DM (sarsaponin; Yucca schidigera extract). These levels were far below thresholds expected to be adverse in ruminants. Effects were measured in lambs by comparison with unsupplemented control diets calculated to be either deficient (10%) or adequate in protein. The diets consisted of hay, concentrate (1:1) and extra wheat starch with increasing body weight. Ruminal pH, VFA concentration, protozoa count and apparent digestibilities of organic matter and fibre did not differ among treatments. The low tannin dose significantly decreased bacteria count compared to the high saponin dose. Saponin supplementation and the high tannin dose showed some potential to reduce ruminal ammonia concentration. This was associated with weak trends towards lower urine N excretion (only tannins) and ammonia emission from manure. Methane release was increased by the low tannin dose compared to the unsupplemented control. Diet effects on heat production were not systematic. In conclusion, the extracts rich in tannins or saponins gave only slight indications for either increased body nitrogen retention or reduced nitrogen emission. However, effects might have been larger with more pronounced dietary protein deficit.     

The Human W42R γD-Crystallin Mutant Structure Provides a Link between Congenital and Age-related Cataracts

Some mutants of human γD-crystallin are closely linked to congenital cataracts, although the detailed molecular mechanisms of mutant-associated cataract formation are generally not known. Here we report on a recently discovered γD-crystallin mutant (W42R) that has been linked to autosomal dominant, congenital cataracts in a Chinese family. The mutant protein is much less soluble and stable than wild-type γD-crystallin. We solved the crystal structure of W42R at 1.7 Å resolution, which revealed only minor differences from the wild-type structure. Interestingly, the W42R variant is highly susceptible to protease digestion, suggesting the presence of a small population of partially unfolded protein. This partially unfolded species was confirmed and quantified by NMR spectroscopy. Hydrogen/deuterium exchange experiments revealed chemical exchange between the folded and unfolded species. Exposure of wild-type γD-crystallin to UV caused damage to the N-terminal domain of the protein, resulting in very similar proteolytic susceptibility as observed for the W42R mutant. Altogether, our combined data allowed us to propose a model for W42R pathogenesis, with the W42R mutant serving as a mimic for photodamaged γD-crystallin involved in age-related cataract.     

Multiscale structures of lipids in foods as parameters affecting fatty acid bioavailability and lipid metabolism

On a nutritional standpoint, lipids are now being studied beyond their energy content and fatty acid (FA) profiles. Dietary FA are building blocks of a huge diversity of more complex molecules such as triacylglycerols (TAG) and phospholipids (PL), themselves organised in supramolecular structures presenting different thermal behaviours. They are generally embedded in complex food matrixes. Recent reports have revealed that molecular and supramolecular structures of lipids and their liquid or solid state at the body temperature influence both the digestibility and metabolism of dietary FA. The aim of the present review is to highlight recent knowledge on the impact on FA digestion, absorption and metabolism of: (i) the intramolecular structure of TAG; (ii) the nature of the lipid molecules carrying FA; (iii) the supramolecular organization and physical state of lipids in native and formulated food products and (iv) the food matrix. Further work should be accomplished now to obtain a more reliable body of evidence and integrate these data in future dietary recommendations. Additionally, innovative lipid formulations in which the health beneficial effects of either native or recomposed structures of lipids will be taken into account can be foreseen.     

Effect of Evening Exposure to Dim or Bright Light on the Digestion of Carbohydrate in the Supper Meal

In a previous study we found that daytime exposure to bright as compared to dim light exerted a beneficial effect on the digestion of the evening meal. This finding prompted us to examine whether the digestion of the evening meal is also affected by evening light intensity. Subjects lived in light of 200 lux during the daytime (08:00–17:00 h) and took their evening meal at 17:00 h under 20 lux (evening dim‐light condition: 17:00–02:00 h) or 2000 lux (evening bright‐light condition: 17:00–02:00 h) until retiring at 02:00 h. Assessment of carbohydrate digestion of the evening meal was accomplished by a breath hydrogen test that is indicative of the malabsorption of dietary carbohydrate. Hydrogen excretion in the breath in the evening under the dim‐light condition was significantly less than under the bright‐light condition (p < 0.05). This finding is the opposite to that obtained in previous experiments in which subjects were exposed to the different intensities of light during the daytime, and indicates that the exposure to dim light in the evening exerts a better effect on carbohydrate digestion in the evening meal than does the exposure to bright light.     

Preliminary investigation of the digestive enzymes in Pagellus erythrinus (Linneo 1758) larvae

To gain a better understanding of the role of the olfactory system in sexual selection, we conducted a series of behavioral assays to correlate the number of aesthetascs with pre-copulatory behavior and male mating success in a protandrous simultaneous hermaphroditic shrimp, Lysmata wurdemanni. We demonstrated that male pre-copulatory behavior and mating success were closely associated with the number of aesthetascs. Individuals with a higher numbers of aesthetascs started approaching/searching females earlier and had higher mating success than ones with lower numbers. Sexual selection theory predicts that small males in a pure searching mating system have advantages to mate with females because they are cryptic and agile. However, our results show that aesthetasc number, rather than shrimp size, may be the major factor in determining male mating success.