Direct contact method of autoradiography was applied in order to know in Eurytrema pancreaticum which was incubated at various intervals such as 60, 120, 240, and 360 minutes in the vitro media added with radioactive succinic acid, C(14)-succinate. The distribution of the radioactive succinate was clarified by this technique and possible explanation was presented. The autoradiographic procedures were essentially the same as those reported previously by Yoon et al. (1964). The most distinct autoradiograms were obtained after 240 minutes incubation. The radioactivity of this labelled succinic acid chiefly concentrated in reproductive organs, such as testes, ovary, egg containing uterine tubules and vitelline follicles.
paraisology-helminth-trematoda-Eurytrema pancreaticum ; autoradiography ; metabolism-succinic acid ; succinic acid

The glucose uptake rate by Eurytrema pancreaticum was a mean value of 16.44 +/- 2.42 micro-mole/hr/g, and total CO(2) production rate by the fluke averaged 5.82 +/- 0.97 micro-mole/hr/g. The relative specific activity of respiratory CO(2) showed a mean value of 5.75 +/- 0.84 per cent. The rate of CO(2) production derived from medium C(14)-glucose was a mean of 0.33 +/- 0.10 micor-mole/hr/g. Therefore, the average value of 0.32 +/- 0.04 per cent of glucose utilized by the flukes from the medium C(14)-glucose was oxidized to respiratory CO(2). The tissue concentration of glycogen in E. pancreaticum was a mean of 45.50 +/- 2.18 mg/g or 4.55 +/- 0.22 %/g. But the turnover rate of glycogen pool was a mean of 0.027 +/- 0.003 %/hr or 0.009 +/- 0.002 mg/hr/g. The average value of 0.64 +/- 0.23 percent of glucose utilized by the flukes from the medium C(14)-glucose was incorporated into the glycogen. These data account for that only 1 per cent of the utilized glucose by the flukes participated in furnishing the oxidation into respiratory CO(2) and the synthetic process into glycogen.
parasitology-helminht-trematoda- Eurytrema pancreaticum ; glucose-biochemistry ; autoradiograhy ; glucose ; glycogen ; CO(2)

The adult trematodes, Fasciola hepatica, Eurytrema pancreaticum and Paramphistomum cervi, employed in this experiment were obtained from the cattle slaughtered at the local abbatoir. The worms selected and washed several times in normal sterilized saline solution. Each about ten of intact F. hepatica, fourty of E. pancreaticum, and twenty of P. cervi were incubated in 50 cc volume of special incubation flasks with incubation medium consisting of 10 cc. of Krebs-Ringer phosphate buffer(pH 7.4) The incubation medium was added C(14)-1-acetate and non-radioactive carrier Na-acetate so as to contain acetate concentration of 50 mg per cent . The worms were allowed to incubate for 5 hours in the Dubnoff metabolic shaking incubator at 38 C. After incubation period, respiratory CO(2) samples from central well of incubation flask were analysed for total CO(2) production rate and their specific activity of respiratory CO(2). The lactate and pyruvate appearance rates were determined by analyzing the lactate and pyruvate concentration in a medium after incubation. The glycogen samples isolated from worms were analyzed for the tissue concentration and their radioactivities in order to determine the turnover rate of glycogen pool. Radioactivities of these series of experiments were counted by an endwindow Geiger-Muller counter as an infinitely thin samples. The quantitative analysis of C(14)-acetate utilized by F. hepatica, E. pancreaticum and P. cervi were compared and discussed in this report. According to these data of the experiment, it is suggested that the fatty acid such as acetate may play a part of their oxidative process into the respiratory CO2 and the synthetic process into glycogen in the above species of trematodes.
parasitology ; helminth ; trematoda ; Fasciola hepatica ; Eurytrema pancreaticum ; Paramphistomum cervi ; acetate ; metabolism ; biochemistry ; CO(2) ; glycogen ; Krebs-Ringer phosphate buffer

The life cycle of Eurytrema pancreaticum has been studied locally at Chejudo (Quelpart Island) in Korea, and found a land snail, Acusta despecta Gray served as the first intermediate host of the pancreatic fluke. The second intermediate host of the fluke also has been surveyed there, the tettigoniid grasshoppers, Conocephalus maculatus Le Guillon and C. gladiatius Redtenbacher. The land snail, Acusta despecta Gray, is acquired infection with the eggs of the fluke in the autum, then the miracidia grow in the liver of the land snails and become mother sporocysts through the winter. After one mother sporocyst has been divided into many spindle shaped daughter sporocysts during spring session, the fully matured daughter sporocysts penetrate into the membrane of mantle cavity in the land snail, and then, pass actively the membrane between June and July. The daughter sporocysts are eaten by the tettigoniid grasshoppers, C. maculatus and C. gladiatus, through the summer and autumn. Cercariae in the grasshoppers grow for about 20 days, and become matured metacercariae in the abdominal cavity. Finally, the matured metacercariae together with the grasshoppers are eaten by their final hosts; goat, rabbit, etc. The artificial infection with the metacercariae of the pancreatic fluke to rabbit as the final host in laboratory room was carried out successfully. The miracidia have been forced out by pressure from the eggs incubated at 20-25 C could live more than three hours swimming around the medium under the microscope.
parasitology-helminth-trematoda ; Eurytrema pancreaticum ; life cycle ; morphology ; egg ; snail-Acusta despecta ; rabbit ; grasshopper-Conocephalus maculatus ; Conocephalus gladiatius

Direct contact method of autoradiography was utilized in studying the distribution of exogenous C(14)-proline in Clonorchis sinensis, Eurytrema pancreaticum, Hymenolepis diminuta and Dipylidium caninum. The most distinct autoradiogram were obtained after 60 min of incubation, corresponding to the maximal absorption of C(14)-proline in these parasite. The radioactivitity of this labeled amino acid was chiefly concentrated in reproductive organs, especially egg-containing uterine tubules.
parasitology-helminth-trematoda-cestoda ; Clonorchis sinensis ; Eurytrema pancreaticum ; Hymenolepis diminuta ; Dipylidium caninum ; autoradiography ; C(14)-proline ; biochemistry ; amino acid

The adult trematode, Fasciola hepatica and Eurytrema pancreaticum, employed in this experiment were obtained from the cattle slaughtered at the local abbatoir. The worms were selected and washed several times in normal sterilized saline solution. Each ten of intact F. hepatica and about thirty to fifty of E. pancreaticum were incubated in 50 cc volume of special incubation flasks with incubation medium consisting of 50 cc of Krebs-Ringer phosohate buffer (pH 7.4). The incubation medium was added C(14)-lactate and non-radioactive carrier Na-lactate so as to contain lactate concentration of 32 mg per cent. The worms were allowed to incubate for 3 hours in the Dubnoff metabolic shaking incubator at 38 C. After incubation period, respiratory CO(2) samples from central wall of incubation flask were analysed for total CO(2) production rate and their specific activity of respiratory CO(2). The lactate uptake rate was determined by analyzing the the difference between lactate concentration in a medium before and after the incubation period, and the pyruvate appearance rate was dertermined by analyzing the pyruvate concentration in a medium after incubation. The glycogen samples isolated from worms were analyzed for the tissue concentration and their radioactivities in order to determine the turnover rate of glycogen pool. Radioactivities of these serise of experiment were counted by an endwindow Geiger-Muller counter as an infinitely thin samples. The quantative analysis of C(14)-lactate utilized by F. hepatica and E. pancreaticum were summerized and compared as following. In F. hepatica the lactate uptake rate was a mean value of 1.04+/-0.15 micromole/hr/g of wet wt. and pyruvate apperance rate was a mean value of 0.132+/-0.005 micro-mole/hr/g of wet wt. The total CO(2) production rate by the flukes averaged 13.82+/-0.75 micro-mole/hr/g of wet wt. The relative specific activities of respiratory CO(2) showed a mean value of 9.93+/-0.62 per cent. The rate of CO(2) production derived from medium C(14)-lactate was a mean of 1.38+/-0.13 micro-mole/hr/g of wet wt. Therefore the averge value of 55.27+/-5.78 per cent (R.L.D. CO(2)) and 15.35+/-1.90 per cent (R.L.D. pyr) of lactate was oxidized into respiratory CO(2) and pyruvate respectively. On the other hand, in E. pancreaticum the lactate uptake rate was a mean value of 0.61+/-0.18 micromole/hr/g of wet wt, and pyruvate appearance rate was a mean of 0.023+/-0.001 micromole/hr/g of wet wt. The total CO(2) production rate by the E. pancreaticum averaged 4.29+/-0.85 micromole/hr/g of wet wt. The relative specific activity of respiratory CO(2) (R.S.A CO2) showed a mean value of 9.20+/-0.34 per cent. Thus, a mean value of 9.20 per cent of total CO(2) production rates was originated from C14-lactate in a medium, therefore the rate of CO(2) production derived from medium C(14)-lactate was a mean value of 0.40+/-0.10 micromole/hr/g of wet wt. The average value of 23.93+/-7.11 per cent(R.L.D. CO(2)) and 3.86+/-0.45 per cent(R.L.D. pyr) of lactate was oxidized into respiratory CO(2) and pyruvate respectively. The tissue concentration of glycogen in F. hepatica was a mean of 2.63 per cent/g of wet wt, while in E. pancreaticum was a mean of 4.06 per cent/g of wet wt. The turnover rate of glycogen pool in F. hepatica yielded a value of 0.073+/-0.008 micromole/hr/g of wet wt whereas in E. pancreaticum yielded only a mean of 0.006+/-0.002 mg/hr/g of wet wt. Therefore, the half time of glycogen turnover, which is the time interval required to replace the half of glycogen pool with medium C(14)-lactate, gave value of a mean of 10.73+_0.76 days in F. hepatica. However, incorporation of C(14)-lactate into glycogen was negligible in the E. pancreaticum. Theses data impressed that the carbohydrate such as lactate may play a role of major part of their oxidative metabolism in F. hepatica, whereas minor part of lactate participates in the oxidative metabolism in E. pancreaticum.
parasitology-helminth-trematoda ; Fasciola hepatica ; Eurytrema pancreaticum ; autoradiography ; biochemistry ; pyruvate ; lactate ; glycogen ; metabolism ; Krebs-Rigner phosphate buffer

In order to obtain some informations on the nature and relative activity of the phosphatases present in various helminths, biochemical studies have been made in thirteen kinds of worm parasites including the adults and larvae (Fasciola hepatica, Eurytrema pancreaticum, Paramphistomum sp., Taenia solium, Taenia pisiformis, Dipylidium caninum, Diphyllobothrium mansoni, Cysticercus cellulosae, Cysticercus fasciolaris and Sparganum). A comparison based on the analysis of pH-activity curves was made among these helminths. The worm materials were mostly obtained alive from an abattoir and removed from the organs or tissues of the animal hosts naturally infected. Sparganum and Cysticercus cellulosae, however, are collected from the subcutaneous tissue of the patients by surgical removal. The worms thoroughly washed were weighed and transferred with 0.1 M Tris buffer to a chilled glass grinder (Capacity; 15 ml) and homogenized in the cold. The homogenate was centrifuged at 5000 RPM for 30 minutes. The supernatant was pipetted off for determination of the phosphatase activity. Incubation mixtures consisted of 1 ml substrate, 1 ml buffer and 0.5ml extract. The buffers used were Tris (Hydroxymethyl) aminomethane and citric acid monohydrate and the substrate was paranitrophenyl phosphate (1 gm/25 ml). These mixtures were incubated at the temperature of 37 C for 30 minutes in water bath. The absorbance or transferance of mixture was determined colorimetrically by "Spectronic 20 "spectrophotometer at 410 nm against a distilled water blank. The amount of phenol liberated was then calculated from a standard curve using phenol solutions. Controls consisted of unincubated mixtures. The results were deducted from this experiment. The phosphatase activity occurred over all parasitic helminths used in this experiment. In trematodes, pH-activity curves have demonstrated two peaks of phosphatase activity in Fasciola hepatica and Paramphistomum species. However the acid phosphatase activity was predominantly found and the alkaline phosphatase activity was found distinctly to be low in all three species. In Eurytrema pancreaticum, the pH-activity curves displayed two peaks in acid phosphatase activity, one at pH 5.0 and the other pH 9.0. In cestodes, both alkaline and acid phosphatase activity displayed the pH optima 5.0 and 9.0 to 10.0 in the adult tapeworms. However, major activity in the adults is due to the alkaline phosphtases. In contrast to the adults, Cysticercus and sparganum showed the higher activity in acid phosphatases which predominates in the larvae. In all cases of nematodes, the pH optimum for acid phosphatase was 4.0 to 6.0. A preponderance of acid phosphatase activity was shown in the extract of intestine of Ascaris lumbricoides. The aspect that phosphatases are correlated with phosphorylated passage of substances through the cuticle of helminths and may also be involved in carbohydrate metabolism is discussed.
parasitology-helminth-trematoda-cestoda ; Fasciola hepatica ; Eurytrema pancreaticum ; Paramphistomum sp. ; Taenia solium ; Taenia pisiformis ; Dipylidium caninum ; Diphyllobothrium mansoni ; Cysticercus cellulosae ; Cysticercus fasciolaris ; sparganum ; alkaline phosphatase ; acid phosphatase ; biochemistry

A comparison of the absorption and incorporation of C(14)-proline into protein by the 7 kinds of helminth parasites is presented. The radioactivity of free amino acid fraction is greater than that of protein fraction in all the worms, and only a small amount of exogenous labeled proline is incorporated into tissue protein. In general, the pattern of C(14)-proline uptake and its incorporation into protein shows rapid linear increase during the period of 15 to 30 min and reaches the maximum at 60 min after incubation, and then the equilibrium state was maintained throughout further incubation.
parasitology-trematoda-cestoda-helminth ; Fasciola hepatica ; Eurytrema pancreaticum ; Metastrongylus longatus ; Hymenolepis diminuta ; Diphyllobothrium caninum ; sparganum ; sparganosis ; metabolism ; biochemistry ; radioactivity ; amino acid ; C(14)-proline ; protein

The mineral contents of the adult Clonorchis sinensis from rabbits and human were measured, and its qualitative and semi-quantitative analyses were studied with 3.4 meter Ebert-Grating spectrograph, and its quantitative analyses were studied with Beckman DU spectrophotometer. The adult Clonorchis sinensis used in this study were divided into two groups, the first group was collected from the bile passage of the man (C. sinensis from man), the second group was collected from the bile passage of the rabbit(C. sinensis from rabbit). Simultaneously, spectrographic and photometric analyses were also performed on the adult worms of Paragonimus westermani. Furthermore, comparative spectrographic analyses of the trace elements were carried out on the C. sinensis from man, C. sinensis from rabbit, Fasciola hepatica, Eurytrema pancreaticum, and Paragonimus westermani, and the approximate contents of the trace elements of the above trematodes were compared with that of their host tissues and biles. The results obtained were as follows: In the spectrographic analyses of C. sinensis from man, sodium, phosphorus, potassium, copper, calcium, aluminum, iron, and magnesium were detected predominantly, and zinc, titanium, silicone, lead, manganese, barium, cromium, molybdenum, and silver were detected as trace elements. In the differences of level of the minerals in these two C. sinensis, copper level of C. sinensis from man was higher than that of C. sinensis from rabbit, while calcium level of the former worm was higher than that of the latter worm. The trace level of lead, molybdenum, and silver were acertained in the former, but latter were not detected . The contents of the minerals showed the characteristic features in each trematodes: the mineral of flukes in each host were much more than that of the others; such as copper in C. sinensis from man, calcium in the C. sinensis from rabbit, and silicone in the P. westermani. The vanadium was detected in the F. hepatica, E. pancreaticum, and P. westermani, while the other flukes were not detected. In the comparative spectrographic analyses of the trace elements among the trematodes and their host tissues and biles, the minerals which detected from flukes were also found in their tissues and biles of their host. But the mineral levels of C. sinensis from man, F. hepatica, E. pancreaticum, and P. westermani were higher than that of their host tissues and biles, except the C. sinensis from rabbit.
parasitology ; helminth ; trematoda ; C. sinensis ; rabbit ; Fasciola hepatica ; Eurytrema pancreaticum ; Paragonimus westermani ; biochemistry ; sodium ; phosphorus ; potassium ; copper ; calcium ; aluminum ; iron ; magnesium ; zinc ; titanium ; silicone ; lead ; manganese ; barium ; cromium ; molybdenum ; silver ; vanadium

By an application of Sigma-Frankel methods, two transaminase systems, glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase, were found to operate at a mesurable rate in 2 species of nematodes(Ascaris lumbricoides and Ascaridia galli), 5 species of trematodes (Clonorchis sinensis, Fasciola hepatica, Eurytrema pancreaticum, Paramphistomum cervi and Paragonimus westermani) and 5 kinds of cestodes (Diphyllobothrium mansoni, Dipylidium caninum, Taenia pisiformis, Cysticercus cellulosae and Cysticercus pisiformis). A comparison was made of the transamination reactions in nematodes and those of trematodes and cestodes. And the significance of transaminase in these parasites is discussed in relation to protein synthesis and its utilization.
parasitology-helminth-nematoda-trematoda-cestoda ; transaminase ; biochemistry ; spectrophotometry ; Ascaris lumbricoides ; Ascaridia galli ; Clonorchis sinensis ; Fasciola hepatica ; Eurytrema pancreaticum ; Paramphistomum cervi ; Paragonimus westermani ; Diphyllobothrium mansoni ; Dipylidium caninum ; Taenia pisiformis ; Cysticercus cellulosae ; Cysticercus pisiformis