Fasciola gigantica: Histology of the digestive tract and the expression of cathepsin L

The digestive tract of Fasciola gigantica is composed of the oral sucker, buccal tube, pharynx, esophagus, and caecum. The tegumental-type epithelium lines the first four parts of the digestive tract while the caecal-type epithelium lines the remaining parts from the caecal bifurcation. The caecal-epithelial cells are classified into 3 types according to their staining properties and ultrastructural characteristics, as related to the amount of food contents in the caecal lumen. All caecal-type epithelial cells synthesize and secrete cathepsin L, a major group of enzymes in the digestive tract, as detected by in situ hybridization and immunolocalization. Moreover, the secreted cathepsin L is also adsorbed on the outer surface of the tegument and the glycocalyx coating of the surface of the tegument, whereas the tegumental cells and tegumental syncytium covering the parasite’s body and lining the proximal part of the digestive tract exhibit no in situ hybridization signal and immunostaining for cathepsin L.     

Responses of midgut amylases of Helicoverpa armigera to feeding on various host plants

Midgut digestive amylases and proteinases of Helicoverpa armigera, a polyphagous and devastating insect pest of economic importance have been studied. We also identified the potential of a sorghum amylase inhibitor against H. armigera midgut amylase. Amylase activities were detected in all the larval instars, pupae, moths and eggs; early instars had lower amylase levels which steadily increased up to the sixth larval instar. Qualitative and quantitative differences in midgut amylases of H. armigera upon feeding on natural and artificial diets were evident. Natural diets were categorized as one or more members of legumes, vegetables, flowers and cereals belonging to different plant families. Amylase activity and isoform patterns varied depending on host plant and/or artificial diet. Artificial diet-fed H. armigera larvae had comparatively high amylase activity and several unique amylase isoforms. Correlation of amylase and proteinase activities of H. armigera with the protein and carbohydrate content of various diets suggested that H. armigera regulates the levels of these digestive enzymes in response to macromolecular composition of the diet. These adjustments in the digestive enzymes of H. armigera may be to obtain better nourishment from the diet and avoid toxicity due to nutritional imbalance. H. armigera, a generalist feeder experiences a great degree of nutritional heterogeneity in its diet. An investigation of the differences in enzyme levels in response to macronutrient balance and imbalance highlight their importance in insect nutrition.     

Phenolic oxidase activities in glandular trichomes of Solanum berthaultii

Polyphenol oxidase and peroxidase activities were detected in foliar glandular trichomes of the wild, insect-resistant potato species, Solanum berthaultii. These enzyme activities may provide the basis for conversion of clear, viscous trichome exudate into a hard, brown substance which is formed rapidly after insect attack.     

Extracytoplasmic phosphatases of selected species of Saccharomyces, Kluyveromyces and Rhodotorula

Phosphatase activities of yeasts belonging to the genera Saccharomyces, Kluyveromyces and Rhodotorula were studied. Rhodotorula rubra exhibited activities at acid, neutral and alkaline pH; the other yeasts only had activity at acid pH. Growing yeasts in a constant pH (4.5) medium decreased phosphatase activities in Saccharomyces and Kluyveromyces, while neutral activity was enhanced in Rhodotorula rubra which excreted more enzyme under these conditions. Washing cells with sucrose solutions lowered phosphatase activities in all yeasts, due to enzyme liberation. Acid phosphatase activities in isolated and purified cell walls were very small. Phosphatases thus appear not to be strongly bound to yeast cell walls.     

The peritrophic membrane and cocoon ribbons in larvae of Cionus scrophulariae

Feeding larvae of Cionus scrophulariae secrete a diffuse peritrophic membrane (PM). The principal site of production is the posterior mid-gut. At the pre-pupal stage the mid-gut switches its chitinogenic activity from PM production to synthesising ribbons of striking appearance. These ribbons, which are used for cocoon construction, are formed in deep crypts which occur only in the posterior mid-gut. Microfibrillar material for both PM and cocoon ribbons is formed at the tips of the microvilli. The e.m. observations of type I PM formation in at least Cionus indicate that there is not sufficient justification for the general acceptance of the theories of templating and delamination. Similar unsettling evidence obtained by other authors is discussed.     

Antioxidant status of Lobiger serradifalci and Oxynoe olivacea (Opisthobranchia, Mollusca)

Invasion of the Mediterranean Sea by the two world-wide famous exotic algae species, Caulerpa taxifolia and Caulerpa racemosa, is still a problem and has adverse effects on the Mediterranean sublittoral ecosystem. Biological control studies revealed that the two native Sacoglossans, Oxynoe olivacea and Lobiger serradifalci, may have an effect on the expansion of invasive Caulerpa spp. in the Mediterranean. In the framework of this study, antioxidant enzyme activities, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), lipid peroxidation (LPO) and oxidized glutathione (GSSG) levels, as oxidative stress markers in L. serradifalci and O. olivacea were determined at two different temperature conditions (20 and 27 °C). In both species, SOD, CAT and GSH-Px activities were found to be positively correlated with temperature. The SOD activities in L. serradifalci were higher than those in O. olivacea at both temperatures, whereas the CAT and GSH-Px activities were significantly (p<0.05) higher in O. olivacea, compared to L. serradifalci. As expected, both species showed decreased LPO levels at 27 °C compared to 20 °C. GSSG level at 27 °C in O.olivacea was significantly (p<0.05) higher than that of 20 °C. On the other hand, no statistical (p>0.05) difference in L.serradifalci existed between GSSG levels at two temperatures. But, despite the variations in the antioxidant enzyme activities, there was no significant difference in LPO levels between the species, suggesting that the oxidative consequences of a given environmental condition may vary among different species. Inasmuch as the GSSG levels were in accordance with antioxidant enzyme activities, GSH might have acted as a cofactor of GSH-Px and an individual antioxidant in these sea slugs.     

栉孔扇贝消化系统γ-氨基丁酸的免疫组织化学定位

采用抗生物素-生物素-过氧化物酶法(ABC法)的免疫组织化学技术,对γ-氨基丁酸(GABA)在栉孔扇贝(Chlamys farreri)消化系统各组织器官中的分布进行了定位研究。结果表明,栉孔扇贝的唇瓣、口唇、肠道的上皮细胞均呈GABA阳性反应,阳性反应物质呈颗粒状分布且多集中于上皮细胞游离端;胃上皮中未见GABA阳性反应;肝胰腺的上皮细胞及部分结缔组织中呈现GABA阳性反应,其中部分上皮细胞中的阳性反应物质呈团块状分布。GABA在栉孔扇贝消化系统除胃以外的各器官均有分布,推测其可能参与消化功能的调节作用。     

Enzyme activity of extracellular protein induced in Trichoderma asperellum and T. longibrachiatum by substrates based on Agaricus bisporus and Phymatotrichopsis omnivora

Antagonistic Trichoderma spp. are used throughout the world for the biological control of soil-borne plant diseases. This approach has stimulated an on-going search for more efficient mycoparasitic strains with a high potential for producing extracellular lytic enzymes. This study compares the production of lytic enzymes by native strains of Trichoderma asperellum and Trichoderma longibrachiatum on substrates of differing complexity. The quantity of protein induced by Agaricus bisporus-based medium was higher than that induced by Phymatotrichopsis omnivora-based medium. In P. omnivora medium, T. asperellum exhibited higher chitinolytic and β-1,3-glucanolytic activities than T. longibrachiatum. The enzyme profile was related to the previously reported ability of these strains to inhibit the growth of several soil-borne plant pathogens. NAGase production was similar among the tested indigenous strains of T. longibrachiatum; T479 and T359 produced more endochitinase, T479 produced more glucanase, and T341 and T359 produced more β-1,3-glucanase. The detected variations in glucanase and β-1,3-glucanase activities suggest that the production of these enzymes is strongly influenced by the substrate. Strains T397 and T359 exhibited xylanase activity, which triggers defence mechanisms in plants. Thus, these strains may utilise an additional mechanism of biocontrol.     

DEVELOPMENTAL STUDIES OF THE DIPTERAN SALIVARY GLAND : II. DNase Activity in Chironomus thummi

The deoxyribonuclease (DNase) activity of the dipteran (Chironomus thummi) salivary gland, measured both enzymatically and immunochemically, increases about 7-fold with the onset of metamorphosis. The increase in DNase activity occurs at a time when the activities of other enzymes and the total protein content are decreasing. The increased DNase activity is followed by glandular destruction. It is suggested that the alterations of this activity may be regulated by the activities of specific chromosomal sites, and that the enzyme may, at least in part, account for the glandular destruction observed at the time of increased enzyme activity.     

Observations on the feeding mechanism,structure of the gut,and digestive physiology of the european lobster Homarus gammarus (L.) (Decapoda: Nephropidae)

Investigations have been made on the feeding mechanism, structure of the gut, and digestive physiology of the European lobster Homarus gammarus (L.).Ciné-photography has shown that the mandibles do not possess a masticatory function, merely serving to grip food morsels during the tearing process effected by the pulling action of the third maxillipeds. The remaining maxillipeds, together with the maxillae, then direct food fragments to the mouth for ingestion.Ingestion is facilitated by mucoid secretions discharged from the oesophageal tegumental glands; the glands do not appear to produce any enzymes which directly contribute to the digestive processes.The hepatopancreas is the principal organ concerned with digestion. It possesses a complex tubular organization in which sequential cellular differentiation culminates in the discharge of enzymes from the B-cells for extracellular digestion in the cardiac stomach. The enzymes are synthesized within vacuoles contained in the B-cell precursors (F-cells) and are secreted in three bursts of activity at 0–15 min, 1–2 h, and 3.5–5 h after a meal. The initial secretory phase is holocrine. Extracellular digestion involves esterases, arylamidases, and lipases; endopeptidases have not been positively identified by histochemical means despite the fact that Homarus is a carnivore. There is an intracellular digestive phase, not previously described in decapod crustaceans, at the 7–9 h post-ingestive stage in the hepatopancreatic R-cells which is effected by arylamidases and lipases.Various phosphatase enzymes have been identified in the hepatopancreatic cells. Acid and alkaline phosphatases are apparently concerned with several stages in the digestive cycle, including enzyme synthesis and secretion, and the absorption of digestive products. Adenosine triphosphatase activity is primarily associated with granules located in the distal R-cell cytoplasm; the possible significance of these granules in the elimination of metabolic wastes is discussed. Acid phosphatases and esterases are present in the midgut epithelium. The possibility of a passive uptake of material from the midgut lumen is considered.Faecal material in the hindgut is bound by mucoid secretions derived from the tegumental glands of this alimentary region; the mucus may also assist in defaecation.A complete digestive cycle in Homarus occupies ≈ 12 h.Food reserves in the gut consist principally of fat deposits in the R-cells, but minute amounts of glycogen can also be detected.No evidence of calcium, copper or ferric iron deposition in any part of the alimentary tract was found.     

On the structure of the Z pseudo-organ in Xiphinema diversicaudatum

The structure of the Z pseudo-organ in Xiphinema diversicaudatum consists of a muscular mass of about 80 cells arranged in rings of four cells each. These cells have peripheral nuclei and are connected with a glandular monolayered epithelium, which lines the lumen of the Z pseudo-organ. Globular bodies originate from the membranes of the epithelial tissue. They consist of a series of closed vesicles containing secretions, and of tubules through which the secretions flow into the lumen. The globular bodies have a helicoidal distribution within the lumen of the Z pseudo-organ which presumably slows the passage of the egg. Enzymatic digestion, in vivo, indicates that the secretion consists of acid and basic proteins involved in egg shell formation.     

Strain-differences and inducibility of microsomal oxidative enzymes in Drosophila melanogaster flies

Some basic characteristics of the enzyme system involved in the oxidative metabolism of xenobiotic compounds were investigated in Drosophila melanogaster flies. Attention was focussed on (1) the normal levels of these enzymes and their activities in whole flies, in different parts of the fly's body and in different sexes, (2) the changes in levels and activities of the enzymes elicited by pretreatment of the flies with known enzyme inducers and (3) differences between strains.Four commonly used wild-type (WT) strains, three insecticide resistant strains (IR) and one white-eyed mutant strain were employed. Except in those experiments on sex differences and in spatial distribution in the fly's body of the enzymatic activities, microsomes were isolated from whole-body homogenates of mixtures of female and male flies. Microsomal cytochrome P-450, benzo[a]pyrene (BP) hydroxylation, p-nitroanisole (pNA) demethylation and aminopyrine (AP) demethylation were measured in control flies and in flies pretreated with Aroclor 1254 (AC), phenobarbital (PB) or butylated hydroxytoluene (BHT).In flies of the WT strain Berlin-K, there were no significant differences in BP hydroxylation activity and its inducibility between the two sexes. In males, inducibility of BP hydroxylation activity was similar in the head, thorax and abdomen, but significantly lower in testis. Considerable differences in some enzyme activities were found between the strains. pNA demethylation and AP demethylation were substantially higher in all IR strains, while no correlation could be found between their increased insecticide resistance and BP hydroxylating capacity or cytochrome P-450 content of the microsomes.Response to enzyme inducing compounds was found to be strain-dependent. PB proved to be a more efficient inducer of BP hydroxylation than AC, which does induce pNA demethylation. BHT has inducing properties that are intermediate between PB and AC. IR strain Hikone-R turned out to be an exception, possessing very low BP hydroxylating capacity and a low degree of inducibility of mixed-function oxidase activities. Differential temperature dependence was found for BP hydroxylation as compared with pNA demethylation. While BP hydroxylation was doubled when raising the temperature from 25°C to 35°C, pNA demethylation was reduced by 50%.     

The effect of lectins on the attachment and invasion of Enteromyxum scophthalmi (Myxozoa) in turbot (Psetta maxima L.) intestinal epithelium in vitro

The involvement of the lectin/carbohydrate interaction in the invasion of the turbot intestinal epithelium by Enteromyxum scophthalmi was studied in vitro using explants of turbot intestine and pre-treatment of parasite stages with the plant lectins of Canavalia ensiformis (Con A) and Glycine max (SBA). Both lectins inhibited the attachment and invasion of E. scophthalmi stages to the intestinal epithelium, though the inhibitory effect was higher for SBA than for Con A. Such results point to the involvement of N-acetyl-galactosamine (GalNAc) and galactose (Gal) residues and also of mannose/glucose residues in the E. scophthalmi-intestinal epithelium interaction. The inhibitory effect of both lectins on the parasite adhesion and penetration points to the interest of further studies to confirm the presence of putative lectins recognising GalNAc-Gal and mannose/glucose residues in turbot intestine. The obtained results demonstrated also the adequacy of turbot intestinal explants as an in vitro model to study the interaction with E. scophthalmi.     

Changes in the digestive gland of the loliginid squid Sepioteuthis lessoniana (Lesson 1830) associated with feeding

Changes associated with feeding in the histological and cytological structure of the digestive gland of the loliginid squid Sepioteuthis lessoniana were examined, along with the nature of both the intracellular and extracellular enzymes produced by the gland. The timing of the release of the extracellular enzymes during the digestive cycle was also determined using a quantitative experimental program. Like that of all coleoid cephalopods, the digestive gland was characterised by one type of cell with several functional stages. As is the case for other loliginid squids, however, the digestive cells did not contain the large enzyme-carrying boules that characterise the digestive glands of most cephalopods. Instead, smaller secretory granules were found in the digestive cells and these may be the enzyme carriers. The prominent rough endoplasmic reticulum, large mitochondria and active Golgi complexes present in the digestive cells are characteristic of cephalopods and indicate a high metabolic activity. Like that of other cephalopods, endocytotic absorption of nutrients and intracellular digestion occurs in the digestive gland of this squid. From quantitative and qualitative examinations of structural changes in the digestive gland of S. lessoniana after feeding, a schedule of its function during the course of digestion was proposed. This indicated that digestion was very rapid, being completed in as little as 4 h in S. lessoniana. Extracellular digestive enzymes were only released after the first hour following feeding, which implies that they are stored in the stomach between meals to increase digestive efficiency.     

Effects of the infestation by Actinote thalia pyrrha (Fabricius) on the physiological indexes of Mikania micrantha leaves

Mikania micrantha is a dangerous exotic weed now found throughout Guangdong Province, China. Introduced by the Indonesian Oil Palm Institute, lepidopteran defoliator Actinote thalia pyrrha (Fabricius) is a potential biological control factor for M. micrantha. Changes in the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), polyphenoldoxidase (PPO), the total phenolics content, and the capacity of scavenging 1,1-diphenyl-2-picrylhydazyl (DPPH) free radicals in M. micrantha leaves were analyzed after infestation by A. thalia pyrrha larvae. The results showed that a slight change of all the indexes in the damaged leaves appeared within 3 h. After four days of infestation by A. thalia pyrrha , SOD and POD activities in the damaged leaves were higher than those of the control. But CAT activity was lower than that of the control, indicating that the SOD and POD functions were impeded more than those of CAT. The SOD and POD activities reached their highest values after 48 h of infestation by A. thalia pyrrha and then decreased, whereas the highest activity of CAT occurred after 24 h and then decreased. PPO activity fluctuated greatly with its three peak values being 1.83, 1.92, and 2.17 times that of the control. The total phenolics content increased initially and then decreased after 72 h of infestation by A. thalia pyrrha . The capacity of scavenging DPPH was significantly lower than that of the control (p<0.05). SOD activity positively correlated with CAT and POD activities for both damaged and normal leaves. However, the value of the correlation coefficient in the damaged leaves was higher than that in the normal leaves. The total phenolics content also showed a weak positive correlation with PPO activity. It is suggested that the protective response of M. micrantha to infestation by A. thalia pyrrha was short and limited, but resulted in a reduced function of the leaves, disturbed the metabolism in the protective enzyme system, and decreased the antioxidative capacity. M. micrantha was unable to resist the feeding stress by altering the content of the total phenolics.     

On the habits and nature of association of the copepod Pseudomyicola spinosus with the rock oyster Crassostrea glomerata in New Zealand

Both adults and early stages of Pseudomyicola spinosus (Cyclopoidea, Myicolidae) occur on the labial palps as well as in the gut of the rock oyster Crassostrea glomerata. There is evidence to suggest movements of the copepod to and from the gut. Clear evidence of damage to the epithelial walls of the gut is seen in several regions, particularly in confined portions of the stomach near the openings of the ducts of the digestive diverticula. In addition to mechanical injuries, changes from a columnar to a low squamous epithelium have been observed in one instance; an increase of leucocytes has also been observed in another specimen. The evidence suggests that the myicolid copepod is a parasite in the gut of the oyster.     

Separate monophenolase and o-diphenolase enzymes in Triticum aestivum

Monophenolase and o-diphenolase activities of polyphenol oxidase are usually thought to be a part of the same enzyme complex. It has now been demonstrated that the two catalytic activities of the polyphenol oxidase of wheat grains are separable and reside in different enzymes. The electrophoretically separated monophenolase enzyme showed specificity only for monophenol (l-tyrosine) after its elution from acrylamide gels. Further, this enzyme is confined to the endosperm tissue and is undetectable in the embryonic region of the seedling.     

Antioxidant response of the bivalve Pinna nobilis colonised by invasive red macroalgae Lophocladia lallemandii

Invasive species represent a risk to natural ecosystems and a biodiversity hazard. The present work aims to determine the antioxidant enzyme response – superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX), the phase II detoxifying enzyme – glutathione S-transferase (GST) – and markers of oxidative damage – thioredoxin reductase (TR) and malondialdehyde (MDA) – in gills and digestive gland of Pinna nobilis and to study the antioxidant response effects in the bivalve colonised by the invasive macroalgae Lophocladia lallemandii. Colonised specimens were collected in a control area without L. lallemandii and another area completely colonised by L. lallemandii. All enzyme activities were found to be present in gills and digestive gland, with some tissue differences. CAT and SOD activities were higher in gills than digestive gland, whereas GST activity and MDA levels were higher in digestive gland. The presence of L. lallemandii induced a significant increase in the activities of antioxidant enzymes in both gills and digestive gland, except for CAT activity in gills. GST and TR activities were also increased in both tissues, as well as the MDA concentration. We can conclude that the presence of L. lallemandii colonising P. nobilis induces a biological stress and oxidative damage to the fan mussel.     

刀鲚幼鱼消化系统的组织形态学结构

采用光镜和扫描电镜观察长江刀鲚(Coilia nasus)幼鱼消化系统组织形态学结构。结果显示,刀鲚体长,口裂大,含有犬齿状的颌齿和尖锥状的腭齿,具有5对鳃弓,鳃耙长度明显大于鳃丝且表面附着不规则绒毛状细齿;胃呈"Y"型,胃与肠连接处具有16～21个指状环形幽门盲囊;肠为直肠,较短,比肠长为0.241±0.080;肝分为两叶,胰为独立的器官。刀鲚口咽腔为复层鳞状上皮,含有腺体、大量椭圆形黏液细胞、少量杯状细胞及味蕾;胃黏膜都为典型的单层柱状上皮,含有较多由上皮凹陷形成的胃小凹和胃腺;幽门盲囊具有20～25个丰富的褶皱,占满大部分幽门盲囊腔,黏膜层具有微绒毛;中肠黏膜上皮最发达,形成的褶皱细长且连接成网状,单层柱状上皮与复层扁平上皮交替分布。观察结果表明,刀鲚消化系统具有典型肉食性鱼类特征。     

Oxidative stress tolerance in the filamentous green algae Cladophora glomerata and Enteromorpha ahlneriana

Cladophora glomerata (L.) Kütz. and Enteromorpha ahlneriana Bliding are morphologically similar filamentous green algae that are dominants in the upper littoral zone of the brackish Baltic Sea. As these two species co-exist in a continuously fluctuating environment, we hypothesised that they may have different strategies to cope with oxidative stress. This was tested in laboratory experiments through stressing the algae by high irradiance (600 μmol photons PAR m⁻² s⁻¹) at two different temperatures (15 and 26 °C) in a closed system. Thus, oxidative stress was created by high irradiance (photo-oxidative stress) and/or carbon depletion. The extent of lipid oxidative damage, antioxidant enzyme activities and the amount of hydrogen peroxide excreted by the algae to the surrounding seawater medium were measured. The results suggest that the two species have different strategies: the annual C. glomerata could be classified as a more stress-tolerant species and the ephemeral E. ahlneriana as a more stress-susceptible species. Low temperature in combination with high irradiance created less lipid oxidative damage in C. glomerata than in E. ahlneriana, which was probably related to the higher regular activities of the hydrogen peroxide scavenging enzymes catalase and ascorbate peroxidase in C. glomerata, whereas in E. ahlneriana high activities of these enzymes were only obtained after the induction of oxidative stress. Superoxide dismutase activities were similar in both species, but the mechanisms to remove the hydrogen peroxide produced by the action of this enzyme were different: more through scavenging enzymes in C. glomerata and more through excretion to the seawater medium in E. ahlneriana. The high excretion of hydrogen peroxide, possibly in combination with brominated volatile halocarbons, by E. ahlneriana may have a negative effect on epiphytes and may partly explain why this alga is usually remarkably devoid of epiphytes and grazers compared to C. glomerata.