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 series of experiments was performed to determine the lactic dehydrogenase activity of various parasitic helminths. The enzyme activity was determined by the modified method of Wroblewshi and LaDue (1955) using tissue homogenate of 16 kinds of worm parasites. The worms were mostly collected alive from local abattoir and removed from the organ or tissues of the naturally infected animal host and some materials were also obtained from the human host. They were thoroughly washed and homogenized in chilled glass tissue grinder, and then centrifuged. The supernatants were designated as enzyme preparations, and their enzyme activity was measured by spectrophotometry at the wave length of 340 millimicron. In order to know the effects of temperature and substrate concentration on the enzyme activity, the extinction of reduced Coenzyme I(NADH) was measured at the various conditions of incubation temperature and substrate concentration. The results of this experiments were as follows: The lactic dehydrogenase activity occurred over all kinds of parasites used in this study. Most worms of nematodes and trematodes displayed their maximum activity in the range of pH 2.7-3.5, and cestodes revealed their maximum activity in the ranges of both pH 2.7-3.5 and pH 7.4. In nematodes and trematodes, the lactic dehydrogenase activity increased slowly as incubation temperature increases except in the case of Eurytrema pancreaticum, while the activity in cestodes decreased inversely. The lactic dehydrogenase activity increased in proportion to the increase of substrate concentration in most of worm parasites.
parasitology-nematode-trematoda-helminth ; lacticdehydrogenase ; nicotinamide dinucloetide ; sodium pyruvate ; Ascaris lumbricoides ; Ascaridia galli ; Dirofilaria immitis ; Fasciola hepatica ; Eurytrema pancreaticum ; Paramphistomum sp. ; Clonorchis sinensis ; Paragonimus westermani ; Taenia saginata ; Taenia solium ; Taenia pisiformis ; Dipylidium caninum ; Diphyllobothrium mansoni ; sparganum, Cysticercus cellulosae ; Cysticercus fasciolaris ; biochemistry- enzyme

The malic dehydrogenase activity was determined by the modified method of Ochoa (1955) using tissue homogenates of various parasitic helminths. Worm parasites were mostly collected from local abattoir, and removed from the organ or tissues of the naturally infected animal hosts, and some materials were also obtained from the human hosts. The helminths used in this experiment include 3 kinds of nematodes, 5 kinds of trematodes, and 8 kinds of cestodes. They were throughly washed and homogenized in glass tissue grinder in ice chilled water bath, and then centrifuged. The supernatants were designated as enzyme preparations. The hydrogen concentrations of buffer solution were pH 1.4, 2.7, 3.5, 4.2, 5.2, 7.4, 8.2, 9.3, 10.2, 11.6, and enzymatic reaction of this experiment was performed at incubation temperature of 20, 30, 40, and 50 C. The extinction of Nicotinamide Adenosine Dinucleotide (NAD) was measured by spectrophotometry at the wave length of 340 millimicron. The results of the experiment were as follows: The malic dehydrogenase activity occurred over all kinds of parasitic helminths used in this study. And the activity on sparganum turned out to be highest. All helminths displayed their maximum activity in the range of alkaline pH. A comparison of the effects of temperature and substrate concentration on the enzyme activity was made among these helminths. However, no definite relationship among them has been detected. The significance of the existence of this enzyme in the helminths was briefly discussed.
parasitology-helminth-trematoda-cestoda-nematoda ; Fasciola hepatica ; Eurytrema pancreaticum ; Paramphistomum sp. ; Taenia solium ; Taenia pisiformis ; Dipylidium caninum ; Diphyllobothrium mansoni ; Cysticercus cellulosae ; Cysticercus fasciolaris ; sparganum ; Ascaris lumbricoides ; Ascaridia galli ; Dirofilaria immitis ; Paragonimus westermani ; Clonorchis sinensis ; malic dehydrogenase-biochemistry-enzyme ; malic dehydrogenase ; Nicotinamide Adenosine Dinucleotide