Systematic analysis of the Diplostomidae and Strigeidae (Trematoda)

A systematic analysis of the genera in the Diplostomidae and Strigeidae was made using the Proterodiplostomidae as the outgroup. The Proterodiplostomidae was the family with the greatest preponderance of primitive characters and its monophyly was supported by the unique paraprostate gland. However, no character state supported the monophyly of the Diplostomidae sensu Dubois, 1970. That paraphyletic taxon was composed of 3 monophyletic groups: the pseudosuckerless Neodiplostomidae n. fam. had the most primitive character states of the 3 and its monophyly was based on characters in the neascus/neodiplostomulum metacercariae; the Bolbophoridae n. fam., with pseudosuckers, had its monophyly supported by characters present in the newly named prodiplostomulum metacercaria; and the emended Diplostomidae, also with pseudosuckers, had the most derived states and its monophyly was supported by characters present in the diplostomulum. The presence of pseudosuckers united the Bolbophoridae n. fam., the emended Diplostomidae, and the Strigeidae as a monophyletic assemblage. The Strigeidae had the most derived characters of these 3 taxa and its monophyly was supported by characters in the tetracotyle and the cup-shaped forebody and bilobed tribocytic organ of the adult. In general, the adult stages of these strigeoid families showed very conserved morphology and it was the metacercariae that possessed the innovations. The conserved adult morphology was typical of what one might expect if the intramolluscan stages were analyzed. Thus, the data were concordant with the view that the mollusc and vertebrate definitive host were the original hosts to the Digenea and that the second intermediate host and metacercaria were more recently intercalated. More specifically, the phylogeny of these groups suggested that ancestral bisegmented strigeoids originally infected reptiles, they subsequently radiated into birds with which they coevolved extensively, and on 5 separate occasions they radiated into mammals. The radiations into mammals were, by all available evidence, preceded by second intermediate host shifts from fish to amphibians.     

Trematode metacercariae from the cranial cavity of African catfish Clarias gariepinus (Burchell, 1822) from Lake Tana,Ethiopia

Metacercariae of two species of trematodes from the fam. Diplostomidae from the cranial cavity of the African catfish Clarias gariepinus are described. The metacercaria Tylodelphys grandis n. sp. is described for the first time. Data are presented on the infection rate of catfishes in Lake Tana.     

Activity of digestive enzymes of thick-billed guillemot (<Emphasis Type="Italic">Uria lomvia</Emphasis>) and common guillemot (<Emphasis Type="Italic">U. aalge</Emphasis>) invaded by cestodes

Activities of digestive enzymes (proteases, carbohydrases, acid and alkaline phosphatases) are determined in intestinal mucosa of the thick-billed and common guillemots. Comparative analysis of the obtained results is performed for non-infested and for birds infested by cestodes. It has been established that at infestation by cestode Alcataenia armillaris (Cestoda: Tetrabothriidae), activities of carbohydrase and alkaline phosphatase in intestinal mucosa of the thick-billed guillemot decreases. Parasitizing of cestodes Tetrabothrius jaegerskieldi (Cestoda: Tetrabothriidae) in intestine induces a decrease of saccharase activity. There is studied kinetics of desorption of enzymes from digestive-transport surfaces of the bird intestine. Peculiarities of firmness of enzyme fixation are established on the surface of intestinal mucosa of invaded guillemots. According to the obtained data, a decrease of the carbohydrase activities in intestine of infested guillemots is likely to be due to absorption of a part of enzymes hydrolyzing carbohydrates on the surface of cestodes.     

Effect of <Emphasis Type="Italic">Bacillus subtilis</Emphasis> in feed probiotic on the digestion of fish cultured in cages

We studied the activity of the digestive enzymes, qualitative composition, and bacteria genera ratio in the intestinal chyme of carp Cyprinus carpio and sterlet Acipenser ruthenus grown on an artificial diet in cage culture. It was shown that the inclusion of live bacteria Bacillus subtilis in the probiotic ProStor had a positive impact on the size and the structure of the microbial community in intestinal chyme, on the activity of some digestive enzymes, and on the digestibility and the weight gain of carp and sterlet. The total amount of micro-organisms in the carp chyme was by 13% higher and in sterlet it was 3.5 times higher in the experimental groups compared to the control. In the studied species of fish, the increase in the activity of digestive enzymes was accompanied by the decrease in nitrogen, ammonia, and urea. Presumably, it happened because of better absorption of protein and more intensive use of nitrogen metabolites by intestinal microflora for the synthesis of their own biomass. The average daily weight gain of carp and sterlet was 25 and 35% higher, respectively, in the experimental groups than in the control.     

Digestive enzymes during ontogeny of the sturgeon Acipenser naccarii: intestine and pancreas development

The present work examines the temporal appearance and degree of activity of digestive enzymes of pancreatic and intestinal origin (amylase as well as neutral and alkaline proteases) together with the evolution of the ontogenic development of the intestine and pancreas during the first month of free‐living (30 days post‐hatch–[dph]) of the sturgeon Acipenser naccarii. In addition, the influence of exogenous live feeding with Artemia salina on the detection of digestive enzymes was studied. Shown are that in the first life stages of the A. naccarii, from the time of fertilisation to the juvenile stage, the following events related to its digestive physiology occurred: digestive enzymes were detected in the embryo stage within the egg, presumably related to the hatching gland; in the free‐living embryo stage, opening of the mouth was at 10 dpf and protease and amylase digestive activities occurred due to an early differentiation of the pancreas and intestine (12–13 dpf). At this stage, digestive enzymatic activities are partly owing to the live feed offered before disappearance of the yolk reserves (14 dpf); the expulsion of the melanin plug and the appearance of a continuity of the digestion tract (16 dpf) together with the disappearance of the yolk reserves (17 dpf) mark the passage from the free‐living embryo phase to the juvenile stage; finally, from the first month after hatching in this sturgeon species, there is a stabilisation in digestive amylase and protease activities together with a fully developed digestive structure. The contribution of enzymes from prey in the detection of enzymatic activities determined in the fish was beyond doubt; therefore these exogenous enzymes must also have a certain physiological importance. Subsequent work will be needed to clarify the importance of digestive processes of the exogenous enzyme pool from live feed as well as to assess the possibility of shortening the weaning period.     

Carbohydrate metabolism parameters in the thick-billed murre (Uria lomvia) infested with Alcataenia armillaris (Cestoda: Dilepididae)

Some aspects of carbohydrate metabolism in noninfested thick-billed murres (Uria lomvia) and those infested with the tapeworm Alcataenia armillaris (Cestoda: Dilepididae) are reported. The findings demonstrate that the intestinal invasion by A. armillaris causes a drop in the activity of digestive enzymes (glycosidases and sacharase) and decreases blood glucose levels and glycogen content in the liver. The main reasons underlying changes in avian carbohydrate metabolism in cestode invasion are suggested.     

Characterization of Proteases in the Digestive System of Spiny Lobster (<Emphasis Type="Italic">Panulirus interruptus</Emphasis>)

Enzymes responsible for digestion of food protein were evaluated and characterized in red lobster (Panulirus interruptus). Several tissues, organs, and body fluids were analyzed. The same composition of proteases was found in gastric juice, midgut gland, and intestinal contents. Using specific substrates and inhibitors, we indentified several isotrypsins and isochymotrypsins by gel electrophoresis. Protease activity was found at pH 3 and reduced by using pepstatin A. Operational variables of enzymes were characterized for management of future studies and potential biotechnologies. Types and activities of lobster digestive enzymes constitute background information to study the digestive abilities of the organism further, and will lead to understanding nutritional needs and feeding ecology, mainly because decapods display unique morphologic, metabolic, and behavioral changes during their life cycle. Also, such enzymes become alternative tools for use in biotechnologies.     

Developmental changes in digestive physiology of nestling house sparrows, Passer domesticus

Six decades of studies have speculated that digestive capacity might limit avian growth rate or that developmental changes in the gut might determine developmental changes in digestive efficiency. However, there are no studies on digestive enzymes during avian development, except for studies on mainly domestic birds that exhibit the precocial mode of development. We studied alimentary organ masses, intestinal enzyme activities (sucrase, maltase, isomaltase, aminopeptidase-N), and pancreatic enzyme activities (amylase, trypsin, chymotrypsin) during development of a wild passerine bird exhibiting the altricial mode of development. Wild nestling house sparrows were studied immediately after removal from the nest (days 0, 3, 6 of age; day 0=hatch), whereas captives were raised in the laboratory beginning day 3 on a formulated casein/starch-based diet until fledging age (after day 12). Digestive biochemistry was dynamic. Tissue-specific activities of some digestive enzymes continued to increase through fledging, by >10 times in some cases (e.g., sucrase and maltase in midintestine). Total pancreatic amylase activity increased 100 times between hatch and day 12 through a combination of increases in tissue-specific activity and pancreas mass. House sparrows differ from poultry, in whom after about 2 wk of age the specific activity of intestinal and pancreatic digestive enzymes is generally constant or declines during development. The data on intestinal and pancreatic enzymes help explain why digestive efficiency of nestling house sparrows improves with age, and the data seem consistent with the idea that digestive capacity might limit feeding rate and hence growth rate.     

Proteolytic Activity of Microflora Associated with the Digestive-Transport Surfaces of Intestine of the Burbot Lota lota and of Eubothrium rugosum Parasitized in It

The presence of symbiotic microflora associated with the digestive-transport surfaces of the burbot intestine and of Eubothrium rugosum (Cestoda, Pseudophyllidea) parasitizing in it has been shown. The bacteria absorb amino acids from the cultivation medium or release proteolytic enzymes into the medium, depending on its composition. The studied microorganisms act either as competitors of the parasite and host for nutritive substrates or as suppliers of proteolytic enzymes that can be subsequently used by the parasite and host in the digestive processes. The amino acid absorption and release of proteolytic enzymes by the bacteria do not depend on the medium pH. The microorganisms that are more closely associated with the studied surfaces seem to make a greater contribution to digestive processes both of the parasite and of the host, as bacteria present in the intestinal contents are removed faster from the digestive tract.     

The influence of diet on the digestive enzymes of the Amazon fish Matrinchã, Brycon cf. melanopterus*

The adaptation of the activity of the digestive enzymes of Brycon cf. melanopterus (Characidae) to diets were studied. The activity of amylase, trypsin and lipase can be increased respectively with a diet rich in carbohydrate, protein, or fat. Pepsin did not show change in activity.     

Effect of dexamethasone on activity of glycosidases and proreinases of intestine of sterlet <Emphasis Type="Italic">Acipenser ruthenus</Emphasis>

Effect of dexamethasone (2.0 mg/kg) on activity of enzymes hydrolyzing proteinaceous and carbohydrate food components in the intestine of the starlet Acipenser ruthenus is studied. Dexamethasone modifies the activity of proteinases more than of glycosidases. As a rule, the hormone significantly decreases the level of proteolytic and general amylolytic activity of the intestinal mucosa and chyme in comparison with intact fish on the first day into the experiment and increases it on the 7th or 14th day. The dynamics of activity was different in enzymes of different chains (glycosidase and proteinase) and preparations (the mucosa and chyme).     

A novel chitinase from the earthworm,Eisenia andrei

A novel chitinase gene, EaChi, and its expression pattern from the earthworm, E. andrei are demonstrated. Based on a deduced amino acid sequence, in EaChi, two specific domains for GH family 18 are well conserved with two essential amino acid residues for enzyme activity. The phylogenetic analysis shows that earthworm chitinase, EaChi, is evolutionarily close to other lophotrochozoan chitinases. The expression pattern analysis of EaChi indicates that the major expression is localized at intestinal epithelium and epidermis, possibly suggesting that the prime functions of the chitinase activity could be related to not only digestive process but also self-defending immunity as a biochemical barrier to protect the invasion of chitin-containing pathogens, including fungi, nematodes and protozoa.     

Potential probiotics of the Far Eastern trepang <Emphasis Type="Italic">Apostychopus japonicus</Emphasis> producing digestive enzymes

Data were obtained on species diversity of the digestive tract microflora of the Far Eastern trepang Apostychopus japonicus. The probiotic characteristics of microbial isolates were investigated related to their capacity for synthesis of digestive enzymes (amylase, chondroitin sulfatase, chitinase, and alginate lyase). The Pseudomonas stutzeri strain exhibiting high activity of all the investigated enzymes was found to be preferable among the potential probiotics used for cultivation of the Far Eastern trepang. Preparations based on mixed cultures of a Bacillus pumilus strain with high chondroitin sulfatase and chitinase activities with strains of B. coagulans and B. megaterium K13 with high activity of amylases and alginate lyases are also promising.     

Antagonistic Activity of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> ATCC 4356 on the Growth and Adhesion/Invasion Characteristics of Human <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>

The aim of this research was to determine the potential probiotic activity of Lactobacillus acidophilus ATCC 4356 against several human Campylobacter jejuni isolates. The ability to inhibit the pathogen’s growth was evaluated by co-culture experiments as well as by antimicrobial assays with cell-free culture supernatant (CFCS), while interference with adhesion/invasion to intestinal Caco-2 cells was studied by exclusion, competition, and displacement tests. In the co-culture experiments L. acidophilus ATCC 4356 strain reduced the growth of C. jejuni with variable percentages of inhibition related to the contact time. The CFCS showed inhibitory activity against C. jejuni strains, stability to low pH, and thermal treatment and sensitivity to proteinase K and trypsin. L. acidophilus ATCC 4356 was able to reduce the adhesion and invasion to Caco-2 cells by most of the human C. jejuni strains. Displacement and exclusion mechanisms seem to be the preferred modalities, which caused a significant reduction of adhesion/invasion of pathogens to intestinal cells. The observed inhibitory properties of L. acidophilus ATCC 4356 on growth ability and on cells adhesion/invasion of C. jejuni may offer potential use of this strain for the management of Campylobacter infections.     

Activity of digestive enzymes of thick-billed murre (Uria lomvia) and common murre (U. aalga) invaded by cestodes

Activities of digestive enzymes (proteases, carbohydrases, acid and alkaline phosphatases) are determined in intestinal mucosa of the thick-billed and common murres Comparative analysis of the obtained results is performed for non-infected and for birds infested by cestodes. It has been established that at invasion by cestode Alcataenia armillaris (Cestoda: Tetrabothriidae), activities of carbohydrase and alkaline phosphatase in intestinal mucosa of the thick-billed murre decreases. Parasitizing of cestodes Tetrabothrius jaegerskieldi (Cestoda: Tetrabothriidae) in intestine of the common murre induces a decrease of saccharase activity. There is studied kinetics of desorption of enzymes from digestive-transport surfaces of the bird intestine. Peculiarities of firmness of enzyme fixation are established on the surface of intestinal mucosa of invaded murres. According to the obtained data, a decrease of the carbohydrase activities in intestine of infested murres is likely to be due to absorption of a part of enzymes hydrolyzing carbohydrates on the surface of cestodes.     

Distribution of the enzyme activity in the intestine of beluga <Emphasis Type="Italic">Huso huso</Emphasis> and Russian sturgeon <Emphasis Type="Italic">Acipenser gueldenstaedtii</Emphasis> (Acipenseridae)

Data on distribution of digestive enzymes along the intestinal tract of mature specimens of the beluga Huso huso and Russian sturgeon Acipenser gueldenstaedtii are presented. The absence of a distinct proximo-distal gradient of the enzyme activity can be explained by short length of the intestine of sturgeons. Considerable differences are found in the distribution pattern of enzymes along the intestinal tract of beluga and Russian sturgeon. The effect of the method of calculation of the enzyme activity on the data on the proximo- distal gradient is considered.     

Temporal relationship between the daily profiles of gut melatonin,oxidative status and major digestive enzymes in carp Catla catla

The present study demonstrated a temporal relationship between the concentrations of melatonin, oxidative status and digestive physiology in the gut of a tropical carp Catla catla. We measured the levels of gut melatonin, malondialdehyde (MDA) – a faithful marker of intracellular stress, different antioxidants and major digestive enzymes in the carp gut at four different clock hours in a daily cycle under natural photo-thermal conditions. A correlation between the gut variables was sought to point their possible functional relationship. Gut melatonin titers displayed significant diurnal variations with a peak at midday. An identical temporal pattern with the highest value at midday and nadir at midnight was noted in the activities of superoxide dismutase, catalase and glutathione peroxidase. In contrast, levels of MDA and reduced glutathione (GSH) were highest at midnight and lowest at midday. The activity of all the studied digestive enzymes (α-amylase, cellulase, protease and lipase) showed significant daily variations with a peak at midday. Gut melatonin concentrations by showing a positive correlation with the activity of both enzymatic antioxidants and digestive enzymes, and a negative correlation with the levels of GSH and MDA indicated their possible physiological interplay in a daily cycle. Collectively, our study presented the first information on the daily profiles of oxidative stress, different antioxidants and digestive enzymes in the gut tissues of any fish species, and suggested their functional relationship with the concentrations of gut melatonin in carp Catla catla.     

Generation of glucagon‐like peptide‐2‐expressing Saccharomyces cerevisiae and its improvement of the intestinal health of weaned rats

We aimed to assess the feasibility of enhancing the intestinal development of weaned rats using glucagon‐like peptide‐2 (GLP‐2)‐expressing Saccharomyces cerevisiae (S. cerevisiae). GLP‐2‐expressing S. cerevisiae (GLP2‐SC) was generated using a recombinant approach. The diet of weaned rats was supplemented with the GLP2‐SC strain. The average daily gain (ADG), the intestinal morphology and the activities of the digestive enzymes in the jejunum were tested to assess the influence of the GLP2‐SC strain on intestinal development. The proliferation of rat enterocytes was also assessed in vitro. The study revealed that the ADG of the weaned rats that received GLP2‐SC was significantly greater than that of the controls fed a basal diet (Control) and S. cerevisiae harbouring an empty vector (EV‐SC) (P < 0.05) but was equivalent to that of positive control rats fed recombinant human GLP‐2 (rh‐GLP2) (P > 0.05). Furthermore, GLP2‐SC significantly increased villous height (P < 0.01) and digestive enzyme activity (P < 0.05) in the jejunum. Immunohistochemistry analysis further affirmed that enterocyte proliferation was stimulated in rats fed the GLP2‐SC strain, as indicated by the greater number of enterocytes stained with proliferative cell nuclear antigen (P < 0.05). In vitro, the proliferation of rat enterocytes was also stimulated by GLP‐2 expressed by the GLP2‐SC strain (P < 0.01). Herein, the combination of the GLP‐2 approach and probiotic delivery constitute a possible dietary supplement for animals after weaning.     

Immunohistochemical Identification of Met-Enkephalin in Digestive System and Its Effect on Digestive Enzyme Activities of the Scallop <Emphasis Type="Italic">Chlamys farreri</Emphasis>

The study was designed to determine whether methionine-enkephalin (M-ENK) was present in the digestive system of the scallop Chlamys farreri and investigate the effects of M-ENK on the activity of amylase, protease and lipase in the digestive system of C. farreri. The results indicated that M-ENK-like material was present in the epithelium and connective tissue of labial palps, mouth labia, stomach, intestine, rectum, and hepatopancreas of the scallop C. farreri. Moreover, it was also found that many isolated small cells showing M-ENK-like immunoreactivity were scattered in the epithelial layer of intestine, and many isolated big epithelial cells showing M-ENK-like immunoreactivity were scattered in tubules of hepatopancreas of the scallop. The activity levels of amylase and lipase in crystalline style, hepatopancreas and intestine were enhanced at 1 h after injection of exogenous M-ENK into adductor muscle of the scallops, whereas protease activity levels were significantly suppressed. Our report constitutes the first characterization of M-ENK in the digestive system of scallop C. farreri and investigates the effects of M-ENK on the activities of digestive enzymes of mollusk for the first time. The results suggest an involvement of M-ENK in the functional regulation of the digestive system of C. farreri.     

The relationship between the digestive enzymes in arctic char,Salvelinus alpinus (Salmonidae,Osteichthyes) and its ability to survive in extreme environments

Digestive enzymes of chars from seven Austrian lakes have been studied. Trypsin activity ml⁻¹ fluid is proportional to body size but inversely proportional to the amount of food in the digestive tract. The pancreas appears to secrete trypsin ± continuously and the activity measureable in the intestinal fluid seems to depend largely on the gut passage rate. There is no seasonal rhythm of enzyme activity. Amylase activity is very weak. Pepsin activity seems to depend mostly on the dilution of the gastric juice. The results indicate that the steering mechanisms of digestive enzymes in salmonidae are well distinct from other families. This work was part of a PH. D. thesis (Reimer, 1984). This work was part of a PH. D. thesis (Reimer, 1984).